TW202233824A

TW202233824A - Microfluidic device and method of culturing biological cells in the microfluidic device

Info

Publication number: TW202233824A
Application number: TW110136282A
Authority: TW
Inventors: 藍道爾Ｄ二世羅威; 克麗詩汀畢奧蒙特; 阿蕯凡卡盧那卡蘭; 娜塔莉馬可斯; 傑森Ｍ麥克艾文; ＭＰ懷特; J 坦納內維爾; 王鋼鋒; 安德魯Ｗ麥克法蘭; Ｄ馬萊奧; 凱斯Ｊ布林格; 關曉; 凱文 T 查普曼
Original assignee: 美商伯克利之光生命科技公司
Priority date: 2015-04-22
Filing date: 2016-04-22
Publication date: 2022-09-01
Also published as: TWI819383B; EP3286296A4; HK1253068A1; CN114085730A; TWI744230B; EP3286296A1; CN107849505A; TW201643241A; CN107849505B

Abstract

Systems, methods and kits are described for culturing one or more biological cells in a microfluidic device, including provision of nutrients and gaseous components configured to enhance cell growth, viability, portability, or any combination thereof. In some embodiments, culturing a single cell may produce a clonal population in the microfluidic device.

Description

Microfluidic device and method for culturing biological cells in the same

在生物科學及相關領域中，培養一或若干細胞可為有用的。本發明之一些實施例包含用於在一微流體器件中培養一細胞或細胞群組之裝置及程序。In the biological sciences and related fields, culturing one or several cells can be useful. Some embodiments of the present invention include apparatus and procedures for culturing a cell or population of cells in a microfluidic device.

在一態樣中，提供一種用於培養一或多個生物細胞之微流體器件，其包含：一流動區域，其經構形以含有一第一流體培養基之一流動；及至少一生長室，其包含一隔離區域及一連接區域，該隔離區域與該連接區域流體地連接且該連接區域包含至該流動區域之一近端開口，其中該至少一生長室進一步包含經調節以支援該微流體器件內之細胞生長、存活性、可移植性或其等之任何組合之至少一表面。在一些實施例中，該微流體器件之該隔離區域可經構形以含有一第二流體培養基，且其中當該流動區域及該至少一生長室實質上分別填充有該第一流體培養基及該第二流體培養基時，該第二流體培養基之組分可擴散至該第一流體培養基中及/或該第一流體培養基之組分可擴散至該第二流體培養基中，且該第一培養基可實質上不流動至該隔離區域中。在一些實施例中，該微流體器件可進一步包含具有該流動區域之至少一部分之一微流體通道，且其中該至少一生長室之該連接區域可直接通至該微流體通道中。在一些實施例中，可使用支援該微流體器件內之細胞可移植性之一或多個試劑來調節該至少一調節表面。在一些實施例中，可使用包含伸烷基醚部分之一聚合物來調節該至少一調節表面。在其他實施例中，可使用包含糖類部分之一聚合物來調節該至少一調節表面。在一些實施例中，包含糖類部分之該聚合物可包含聚葡萄糖。在其他實施例中，可使用包含胺基酸部分之一聚合物來調節該至少一調節表面。在一些實施例中，該聚合物可為牛血清白蛋白(BSA)或去氧核糖核酸酶1 (DNase 1)。在其他實施例中，可使用包含羧酸部分、磺酸部分、核酸部分或膦酸部分之一聚合物來調節該微流體器件之該至少一調節表面。在一些實施例中，可使用包含羧酸部分、磺酸部分、核酸部分或膦酸部分之一聚合物來調節該微流體器件之該至少一調節表面。在該微流體器件之各種實施例中，該至少一調節表面包含共價鍵聯至該微流體器件之一表面之一鍵聯基團，且該鍵聯基團可鍵聯至經構形以支援該微流體器件內之細胞生長、存活性、可移植性或其等之任何組合之一部分。在一些實施例中，該鍵聯基團可為矽烷氧基鍵聯基團。在其他實施例中，該鍵聯基團可為膦酸酯鍵聯基團。在各種實施例中，該至少一調節表面可包含烷基或氟烷基部分。在一些實施例中，該等氟烷基部分可為全氟烷基部分。在一些實施例中，該等烷基或氟烷基部分可具有大於10個碳之一主鏈長度。該等烷基或氟烷基部分可具有一直鏈結構。在該微流體器件之各種實施例中，該至少一調節表面之該鍵聯基團可直接鍵聯至經構形以支援細胞生長、存活性、可移植性或其等之任何組合之該部分。在其他實施例中，該鍵聯基團可間接鍵聯至經構形以支援細胞生長、存活性、可移植性或其等之任何組合之該部分。在一些實施例中，該鍵聯基團可經由一鍵聯子而間接鍵聯至經構形以支援細胞生長、存活性、可移植性或其等之任何組合之該部分。在一些實施例中，該鍵聯子可包含伸三唑基部分。在其他實施例中，該鍵聯子可包含一或多個伸芳基部分。在一些實施例中，該至少一調節表面可包含糖類部分。在其他實施例中，該至少一調節表面可包含伸烷基醚部分。在其他實施例中，該至少一調節表面可包含胺基酸部分。替代地，該至少一調節表面可包含兩性離子。在進一步實施例中，該至少一調節表面可包含膦酸部分或羧酸部分。在其他實施例中，該至少一調節表面包含胺基或胍部分。在一些其他實施例中，該至少一調節表面可包含：烷基或氟烷基(其包含全氟烷基)部分；單糖或多糖(其可包含(但不限於)聚葡萄糖)；醇類(其包含(但不限於)炔丙醇)；多元醇，其包含(但不限於)聚乙烯醇；伸烷基醚，其包含(但不限於)聚乙二醇；聚電解質(其包含(但不限於)聚丙烯酸或聚乙烯膦酸)；胺基(其包含其衍生物，諸如(但不限於)烷基化胺、羥烷基化胺基、胍鹽及含有一未芳香化氮環原子之雜環基，諸如(但不限於)嗎啉基或哌嗪基)；羧酸，其包含(但不限於)丙炔酸(其可提供羧酸陰離子表面)；膦酸，其包含(但不限於)乙炔基膦酸(其可提供膦酸陰離子表面)；磺酸陰離子；羧基甜菜鹼；磺基甜菜鹼；胺磺酸；或胺基酸。在該微流體器件之各種實施例中，該微流體器件之該至少一調節表面可包含至少一細胞黏著阻斷分子。在一些實施例中，該至少一細胞黏著阻斷分子可破壞肌動蛋白纖維形成，阻斷整合素受體，或減弱細胞與DNA污損表面之結合。在一些實施例中，該至少一細胞黏著阻斷分子可為一含RGD之肽。在其他實施例中，該至少一細胞黏著阻斷分子可為細胞鬆弛素B、一整合素抗體、纖連蛋白(其可包含一小分子或一DNase 1蛋白)之抑制劑。在其他實施例中，該至少一細胞黏著阻斷分子可包含多於一種類型之細胞黏著阻斷分子之一組合。在該微流體器件之各種實施例中，該微流體器件之該至少一調節表面可包含一可裂解部分。在一些實施例中，該可裂解部分可經構形以容許破壞該調節表面，藉此促進該一或多個生物細胞在培養之後之可移植性。在該微流體器件之各種實施例中，該微流體器件之該至少一調節表面可包含哺乳動物血清之一或多個組分。哺乳動物血清之該一或多個組分可包含B27®補充劑、胎牛血清(FBS)或小牛血清(FCS)。在該微流體器件之各種實施例中，該微流體器件可進一步包含具有一介電泳(DEP)構形之一基板。在一些實施例中，具有一DEP構形之該基板可經構形以將一或多個生物細胞引入至該生長室中或將該一或多個生物細胞移出該生長室。可光學地致動該DEP構形。在該微流體器件之各種實施例中，該微流體器件之該至少一調節表面可經構形以穩定於至少約30°C之一溫度處。在該微流體器件之各種實施例中，該微流體器件之該至少一生長室之該隔離區域可具有足以支援細胞擴增至約100個細胞之一範圍之尺寸。在一些實施例中，可在該至少一生長室中維持不超過1×10 ²個生物細胞，且該至少一生長室之體積可小於或等於約2×10 ⁶立方微米。在其他實施例中，可在該至少一生長室中維持不超過1×10 ²個生物細胞，且該至少一生長室之體積可小於或等於約1×10 ⁷立方微米。在該微流體器件之各種實施例中，該器件可進一步包含：至少一入口，其經構形以將該第一流體培養基或該第二流體培養基輸入至該流動區域中；及至少一出口，其經構形以當該第一流體培養基自該流動區域退出時接收該第一流體培養基。在該微流體器件之各種實施例中，該微流體器件可進一步包含在該至少一生長室或其之該隔離區域上方之一可變形蓋區域，藉此按壓該可變形蓋區域施加足以使該生物細胞自該隔離區域輸出至該流動區域之一力。在該微流體器件之各種實施例中，該微流體器件可包含一蓋，其中該蓋之至少一部分可為可透氣的，藉此將一氣體分子源提供至定位於該微流體器件中之一流體培養基。該蓋之該可透氣部分可定位於該至少一生長室上方。在其他實施例中，該蓋之該可透氣部分可定位於該流動區域上方。在其他實施例中，該至少一生長室可包含複數個生長室。在各種實施例中，該一或多個生物細胞可包含複數個生物細胞。在該微流體器件之各種實施例中，該至少一生長室可包含經調節以支援一哺乳動物細胞之細胞生長、存活性、可移植性或其等之任何組合之至少一表面。在其他實施例中，該至少一生長室可包含經調節以支援一免疫細胞之細胞生長、存活性、可移植性或其等之任何組合之至少一表面。在其他實施例中，該免疫細胞可為一淋巴細胞或白血球。在一些其他實施例中，該免疫細胞可為一B細胞、一T細胞、NK細胞、一巨噬細胞或一樹突細胞。在該微流體器件之各種實施例中，該至少一生長室可包含經調節以支援一貼附細胞之細胞生長、存活性、可移植性或其等之任何組合之至少一表面。在該微流體器件之各種實施例中，該至少一生長室可包含經調節以支援一融合瘤細胞之細胞生長、存活性、可移植性或其等之任何組合之至少一表面。在該微流體器件之各種實施例中，該至少一生長室可包含經調節以支援一單一細胞及生物細胞之一對應選殖群落之細胞生長、存活性、可移植性或其等之任何組合之至少一表面。在另一態樣中，提供一種用於在一微流體器件上培養一或多個生物細胞之系統，該系統包含一微流體器件，其具有：一流動區域，其經構形以含有一第一流體培養基之一流動；及至少一生長室，其中該生長室具有經調節以支援該微流體器件中之細胞生長、存活性、可移植性或其等之任何組合之至少一表面。該至少一生長室可包含一隔離區域及一連接區域，該隔離區域與該連接區域流體地連接且該連接區域具有至該流動區域之一近端開口。在一些實施例中，該微流體器件之該隔離區域可經構形以含有一第二流體培養基，且當該流動區域及該至少一生長室實質上分別填充有該第一流體培養基及該第二流體培養基時，該第二流體培養基之組分可擴散至該第一流體培養基中及/或該第一流體培養基之組分可擴散至該第二流體培養基中，且該第一培養基可實質上不流動至該隔離區域中。在一些實施例中，該微流體器件可進一步包含一微流體通道，其包含該流動區域之至少一部分，且其中該至少一生長室之該連接區域可直接通至該微流體通道中。該微流體器件可為如本文中所描述之任何微流體器件，其具有元件之任何者之任何組合。在該系統之各種實施例中，該系統可進一步包含經構形以至少灌注該第一流體培養基之一流量控制器。該控制器經構形以非連續地灌注該至少第一流體培養基。在該系統之各種實施例中，該系統之該微流體器件可進一步包含具有一介電泳(DEP)構形之一基板，其經構形以將一或多個生物細胞引入至該生長室中或將該一或多個生物細胞移出該生長室。可光學地致動該DEP構形。在該系統之各種實施例中，該系統可進一步包含經構形以含有該第一流體培養基之一儲液器，其中該儲液器流體地連接至該微流體器件。該儲液器可經構形以與能夠使該第一流體培養基飽含溶解氣體分子之一氣體環境相通。在該系統之各種實施例中，該系統可進一步包含連接至該微流體器件之至少一入口之一感測器，其中該感測器可經構形以偵測該第一流體培養基之一pH。在該系統之各種實施例中，該系統可進一步包含連接至至少一出口之一感測器，其中該感測器經構形以在該第一流體培養基離開該微流體器件時偵測該第一流體培養基之一pH。在一些實施例中，該感測器可為一光學感測器。在該系統之各種實施例中，該系統可進一步包含經構形以擷取該至少一生長室及含於其內之任何生物細胞之一影像之一偵測器。在一些實施例中，該一或多個生物細胞可包含一或多個哺乳動物細胞。在其他實施例中，該一或多個生物細胞可包含一或多個融合瘤細胞。在其他實施例中，該一或多個生物細胞可包含一或多個淋巴細胞或白血球細胞。替代地，該一或多個生物細胞可包含一或多個貼附細胞。在該系統之各種實施例中，該生長室中之該一或多個生物細胞可為一單一細胞且該群落可為生物細胞之一選殖群落。在另一態樣中，提供一種組合物，其包含：一基板，其具有一介電泳(DEP)構形及一表面；及一調節表面，其共價鍵聯至該基板之該表面之氧化物部分。該組合物可具有式1或式2之一結構，且可具有式1或式2之元素之任何值，如本文中所界定：

式1 式2 在該組合物之一些實施例中，該調節表面可包含共價鍵聯至該表面之該等氧化物部分之一鍵聯基團，且該鍵聯基團可鍵聯至經構形以支援細胞生長、存活性、可移植性或其等之任何組合之一部分。在一些實施例中，該鍵聯基團可為矽烷氧基鍵聯基團。在其他實施例中，該鍵聯基團可為膦酸酯基。在一些實施例中，該鍵聯基團可直接鍵聯至經構形以支援細胞生長、存活性、可移植性或其等之任何組合之該部分。在一些實施例中，該鍵聯基團可間接鍵聯至經構形以支援細胞生長、存活性、可移植性或其等之任何組合之該部分。在一些實施例中，該鍵聯基團可經由連接至一鍵聯子而間接鍵聯至經構形以支援細胞生長、存活性、可移植性或其等之任何組合之該部分。在一些實施例中，該鍵聯基團可經由連接至一鍵聯子之一第一端而間接鍵聯至經構形以支援細胞生長、存活性、可移植性或其等之任何組合之該部分。該鍵聯子可進一步包含一直鏈部分，其中該直鏈部分之一主鏈包括選自矽原子、碳原子、氮原子、氧原子、硫原子及磷原子之任何組合之1個至200個非氫原子。在一些實施例中，該鍵聯子可進一步包含伸三唑基部分。在一些實施例中，該伸三唑基部分可中斷該鍵聯子之該直鏈部分或可連接至該鍵聯子之該直鏈部分之一第二端。在其他實施例中，該直鏈部分之該主鏈可包含伸芳基部分。在各種實施例中，經構形以支援細胞生長、存活性、可移植性或其等之任何組合之該部分包括烷基部分、氟烷基部分、單糖或多糖、乙醇部分、多元醇部分、伸烷基醚部分、聚電解質部分、胺基部分、羧酸部分、膦酸部分、磺酸鹽陰離子部分、羧基甜菜鹼部分、磺基甜菜鹼部分、胺磺酸部分或胺基酸部分。在一些實施例中，該至少一調節表面可包含胺基酸、烷基部分、全氟烷基部分、聚葡萄糖部分及/或伸烷基醚部分。在一些實施例中，該至少一調節表面可包含烷基或全氟烷基部分。在一些實施例中，該等烷基或全氟烷基部分具有大於10個碳之一主鏈長度。在各種實施例中，該調節表面可進一步包含一或多個可裂解部分。該可裂解部分可經構形以容許破壞該調節表面，藉此促進該一或多個生物細胞在培養之後之可移植性。在另一態樣中，提供一種在一微流體器件中培養至少一生物細胞之方法，該微流體器件具有一流動區域(其經構形以含有一第一流體培養基之一流動)及至少一生長室，該方法包括下列步驟：將該至少一生物細胞引入至該至少一生長室中，其中該至少一生長室經構形以具有經調節以支援細胞生長、存活性、可移植性或其等之任何組合之至少一表面；及在至少足以使該至少一生物細胞擴增而產生生物細胞之一群落的一長時間週期內培育該至少一生物細胞。該至少一生長室可包含一隔離區域及一連接區域，該隔離區域與該連接區域流體地連接且該連接區域具有至該流動區域之一近端開口。在一些實施例中，該微流體器件之該隔離區域可經構形以含有一第二流體培養基，且其中當該流動區域及該至少一生長室實質上分別填充有該第一流體培養基及該第二流體培養基時，該第二流體培養基之組分可擴散至該第一流體培養基中及/或該第一流體培養基之組分可擴散至該第二流體培養基中，且該第一培養基可實質上不流動至該隔離區域中。在一些實施例中，該微流體器件可進一步包含具有該流動區域之至少一部分之一微流體通道，且其中該至少一生長室之該連接區域可直接通至該微流體通道中。該微流體器件可為如本文中所描述之任何微流體器件，其具有元件之任何者之任何組合。在該方法之一些實施例中，該至少一調節表面可包含共價鍵聯至該表面之一鍵聯基團，且其中該鍵聯基團進一步鍵聯至經構形以支援該微流體器件內之該一或多個生物細胞之細胞生長、存活性、可移植性或其等之任何組合之一部分。在一些其他實施例中，經構形以支援細胞生長、存活性、可移植性或其等之任何組合之該部分可包含：烷基或氟烷基(其包含全氟烷基)部分；單糖或多糖(其可包含(但不限於)聚葡萄糖)；醇類(其包含(但不限於)炔丙醇)；多元醇，其包含(但不限於)聚乙烯醇；伸烷基醚，其包含(但不限於)聚乙二醇；聚電解質(其包含(但不限於)聚丙烯酸或聚乙烯膦酸)；胺基(其包含其衍生物，諸如(但不限於)烷基化胺、羥烷基化胺基、胍鹽及含有一未芳香化氮環原子之雜環基，諸如(但不限於)嗎啉基或哌嗪基)；羧酸，其包含(但不限於)丙炔酸(其可提供羧酸陰離子表面)；膦酸，其包含(但不限於)乙炔基膦酸(其可提供膦酸陰離子表面)；磺酸陰離子；羧基甜菜鹼；磺基甜菜鹼；胺磺酸；或胺基酸。在一些實施例中，該至少一調節表面可包含烷基或全氟烷基部分。在其他實施例中，該至少一調節表面可包含伸烷基醚部分或聚葡萄糖部分。在該方法之一些實施例中，該方法可包含下列步驟：調節該至少一生長室之至少一表面。在一些實施例中，調節可包含：使用包含一聚合物之一調節試劑來處理該至少一生長室之該至少一表面。在其他實施例中，調節可包含：使用哺乳動物血清之一或多個組分來處理該至少一生長室之至少一表面。在其他實施例中，調節可包含：使用至少一細胞黏著阻斷分子來處理該至少一生長室之至少一表面。在該方法之一些實施例中，將該至少一生物細胞引入至該至少一生長室中可包含：使用具有足以移動該至少一生物細胞之力量之一介電泳(DEP)力。在一些實施例中，使用一DEP力可包含：光學地致動該DEP力。在該方法之一些實施例中，該方法可進一步包含下列步驟：在該培育步驟期間灌注該第一流體培養基，其中該第一流體培養基經由該微流體器件之至少一入口而引入且經由該微流體器件之至少一出口而輸出，其中在輸出之後，該第一流體培養基視情況包括來自該第二流體培養基之組分。在該方法之一些實施例中，該方法可進一步包含下列步驟：在該培育步驟之後，使該調節表面之一或多個可裂解部分裂解，藉此促進該一或多個生物細胞自該生長室或其隔離區域輸出至該流動區域中。在該方法之一些實施例中，該方法可進一步包含下列步驟：使一或多個生物細胞自該生長室或其隔離區域輸出至該流動區域中。在該方法之一些實施例中，該至少一生物細胞可包含一哺乳動物細胞。在該方法之其他實施例中，該至少一生物細胞可包含至少一免疫細胞。在該方法之其他實施例中，該至少一免疫細胞可包含一淋巴細胞或白血球。在該方法之一些其他實施例中，該至少一免疫細胞可包含一B細胞、一T細胞、NK細胞、一巨噬細胞或一樹突細胞。在其他實施例中，該至少一生物細胞可包含一貼附細胞。替代地，該至少一生物細胞可包含一融合瘤細胞。在該方法之一些實施例中，將該至少一生物細胞引入至該至少一生長室中之該步驟可包含將一單一細胞引入至該生長室中，且由該培育步驟產生之生物細胞之該群落可為一選殖群落。在另一態樣中，提供一種用於培養一生物細胞之套組，其包含一微流體器件，該微流體器件具有：一流動區域，其經構形以含有一第一流體培養基之一流動；及至少一生長室，其包含經調節以支援該微流體器件內之細胞生長、存活性、可移植性或其等之任何組合之至少一表面。該至少一生長室可包含一隔離區域及一連接區域，該隔離區域與該連接區域流體地連接且該連接區域具有至該流動區域之一近端開口。該微流體器件可為如本文中所描述之任何微流體器件，其具有元件之任何組合。在一些實施例中，該微流體器件之該至少一調節表面可包含烷基部分、氟烷基部分、單糖或多糖部分、乙醇部分、多元醇部分、伸烷基醚部分；聚電解質部分、胺基部分、羧酸部分、膦酸部分、磺酸鹽部分、羧基甜菜鹼部分、磺基甜菜鹼部分、胺磺酸部分或胺基酸部分。在一些實施例中，該微流體器件之該至少一調節表面包括下列之至少一者：糖類部分、伸烷基醚部分、烷基部分、氟烷基部分或胺基酸部分。在一些實施例中，該等烷基或氟烷基部分具有大於10個碳之一主鏈長度。在該套組之各種實施例中，該微流體器件之該至少一調節表面可包含共價鍵聯至該微流體器件之一表面之一鍵聯基團，且該鍵聯基團可鍵聯至經構形以支援該微流體器件內之該一或多個生物細胞之生長、存活性、可移植性或其等之任何組合之一部分。在一些實施例中，該鍵聯基團可為矽烷氧基鍵聯基團。在其他實施例中，該鍵聯基團可為膦酸酯鍵聯基團。在一些實施例中，該鍵聯基團可直接鍵聯至經構形以支援細胞生長、存活性、可移植性或其等之任何組合之該部分。在其他實施例中，該鍵聯基團可間接鍵聯至經構形以支援細胞生長、存活性、可移植性或其等之任何組合之該部分。該鍵聯基團可經由一鍵聯子而間接鍵聯至經構形以支援細胞生長、存活性、可移植性或其等之任何組合之該部分。該鍵聯基團可經由連接至一鍵聯子之一第一端而間接鍵聯至經構形以支援細胞生長、存活性、可移植性或其等之任何組合之該部分。在各種實施例中，該鍵聯子可進一步包含一直鏈部分，其中該直鏈部分之一主鏈包括選自矽原子、碳原子、氮原子、氧原子、硫原子及磷原子之任何組合之1個至200個非氫原子。在一些實施例中，該鍵聯子可進一步包含伸三唑基部分。在該套組之各種實施例中，該套組可進一步包含一表面調節試劑。在一些實施例中，該表面調節試劑可包含一聚合物，其包括下列之至少一者：伸烷基醚部分、羧酸部分、磺酸部分、膦酸部分、胺基酸部分、核酸部分或糖類部分。在一些實施例中，該表面調節試劑可包含一聚合物，其包含下列之至少一者：伸烷基醚部分、胺基酸部分及/或糖類部分。在其他實施例中，該表面調節試劑可包含至少一細胞黏著阻斷分子。在一些實施例中，該至少一細胞黏著阻斷分子可破壞肌動蛋白纖維形成，阻斷整合素受體，或減弱細胞與DNA污損表面之結合。在一些實施例中，該至少一細胞黏著阻斷分子可為細胞鬆弛素B、一含RGD之肽、纖連蛋白之一抑制劑、一整合素抗體或一DNase 1蛋白。在一些實施例中，該表面調節試劑可包含一個以上細胞黏著阻斷分子之一組合。在其他實施例中，該表面調節試劑可包含哺乳動物血清之一或多個組分。在一些實施例中，該哺乳動物血清可為胎牛血清(FBS)或小牛血清(FCS)。在該套組之各種實施例中，該套組可進一步包含一培養基添加物，其包含經構形以加強生長室之該至少一表面之該調節之一試劑。該培養基添加物可包含一Pluronics®聚合物。在該套組之各種實施例中，該微流體器件之該至少一調節表面可包含一可裂解部分。在一些實施例中，該套組可進一步包含經構形以使該調節表面之該可裂解部分裂解之一試劑。在該套組之各種實施例中，該套組可進一步包含用以偵測該生物細胞之一狀態之至少一試劑。 In one aspect, a microfluidic device for culturing one or more biological cells is provided, comprising: a flow region configured to contain a flow of a first fluid medium; and at least one growth chamber, It includes an isolation region and a connection region, the isolation region is fluidly connected to the connection region and the connection region includes a proximal opening to the flow region, wherein the at least one growth chamber further includes conditioning to support the microfluidics At least one surface of cell growth, viability, transplantability, or any combination thereof within the device. In some embodiments, the isolation region of the microfluidic device can be configured to contain a second fluidic medium, and wherein the flow region and the at least one growth chamber are substantially filled with the first fluidic medium and the at least one growth chamber, respectively In the second fluid medium, components of the second fluid medium may diffuse into the first fluid medium and/or components of the first fluid medium may diffuse into the second fluid medium, and the first medium may There is substantially no flow into the isolation region. In some embodiments, the microfluidic device can further comprise a microfluidic channel having at least a portion of the flow region, and wherein the connecting region of the at least one growth chamber can directly communicate into the microfluidic channel. In some embodiments, the at least one conditioning surface can be conditioned using one or more agents that support cell portability within the microfluidic device. In some embodiments, the at least one conditioning surface can be conditioned using a polymer comprising an alkylene ether moiety. In other embodiments, the at least one conditioning surface can be conditioned using a polymer comprising a carbohydrate moiety. In some embodiments, the polymer comprising saccharide moieties may comprise polydextrose. In other embodiments, the at least one conditioning surface can be conditioned using a polymer comprising amino acid moieties. In some embodiments, the polymer can be bovine serum albumin (BSA) or deoxyribonuclease 1 (DNase 1). In other embodiments, the at least one conditioning surface of the microfluidic device can be conditioned using a polymer comprising a carboxylic acid moiety, a sulfonic acid moiety, a nucleic acid moiety, or a phosphonic acid moiety. In some embodiments, the at least one conditioning surface of the microfluidic device can be conditioned using a polymer comprising one of a carboxylic acid moiety, a sulfonic acid moiety, a nucleic acid moiety, or a phosphonic acid moiety. In various embodiments of the microfluidic device, the at least one conditioning surface comprises a linking group covalently bonded to a surface of the microfluidic device, and the linking group can be linked to a linking group configured to Part of supporting cell growth, viability, transplantability, or any combination thereof within the microfluidic device. In some embodiments, the linking group can be a siloxy linking group. In other embodiments, the linking group can be a phosphonate linking group. In various embodiments, the at least one conditioning surface may comprise alkyl or fluoroalkyl moieties. In some embodiments, the fluoroalkyl moieties can be perfluoroalkyl moieties. In some embodiments, the alkyl or fluoroalkyl moieties can have a backbone length of greater than 10 carbons. These alkyl or fluoroalkyl moieties may have a straight chain structure. In various embodiments of the microfluidic device, the linking group of the at least one modulating surface can be directly linked to the portion that is configured to support cell growth, viability, transplantability, or any combination thereof, etc. . In other embodiments, the linking group can be indirectly linked to the moiety that is configured to support cell growth, viability, transplantability, or any combination thereof. In some embodiments, the linking group may be indirectly linked via a linker to the moiety configured to support cell growth, viability, transplantability, or any combination thereof. In some embodiments, the linker may comprise a triazolyl moiety. In other embodiments, the linker may comprise one or more arylidene moieties. In some embodiments, the at least one conditioning surface may comprise carbohydrate moieties. In other embodiments, the at least one conditioning surface can include alkylene ether moieties. In other embodiments, the at least one conditioning surface may comprise amino acid moieties. Alternatively, the at least one conditioning surface may comprise zwitterions. In further embodiments, the at least one conditioning surface may comprise phosphonic acid moieties or carboxylic acid moieties. In other embodiments, the at least one conditioning surface comprises amine groups or guanidine moieties. In some other embodiments, the at least one conditioning surface may comprise: alkyl or fluoroalkyl (which includes perfluoroalkyl) moieties; monosaccharides or polysaccharides (which may include, but are not limited to, polydextrose); alcohols (which includes, but is not limited to, propargyl alcohol); polyols, which include, but are not limited to, polyvinyl alcohol; alkylene ethers, which include, but are not limited to, polyethylene glycols; polyelectrolytes (which include ( but not limited to) polyacrylic acid or polyvinylphosphonic acid); amine groups (including derivatives thereof such as, but not limited to, alkylated amines, hydroxyalkylated amine groups, guanidine salts, and containing an unaromatic nitrogen ring Heterocyclyl groups of atoms, such as, but not limited to, morpholinyl or piperazinyl); carboxylic acids, including but not limited to propyolic acid (which can provide a carboxylate anion surface); phosphonic acids, including ( but not limited to) ethynylphosphonic acid (which can provide a phosphonic acid anion surface); sulfonic acid anion; carboxybetaine; sulfobetaine; sulfamic acid; or amino acid. In various embodiments of the microfluidic device, the at least one modulating surface of the microfluidic device can comprise at least one cell adhesion blocking molecule. In some embodiments, the at least one cell adhesion blocking molecule disrupts actin fibril formation, blocks integrin receptors, or reduces cell binding to DNA-fouling surfaces. In some embodiments, the at least one cell adhesion blocking molecule can be an RGD-containing peptide. In other embodiments, the at least one cell adhesion blocking molecule may be an inhibitor of cytochalasin B, an integrin antibody, fibronectin (which may comprise a small molecule or a DNase 1 protein). In other embodiments, the at least one cell adhesion blocking molecule may comprise a combination of more than one type of cell adhesion blocking molecule. In various embodiments of the microfluidic device, the at least one conditioning surface of the microfluidic device can comprise a cleavable moiety. In some embodiments, the lysable moiety can be configured to allow destruction of the regulatory surface, thereby promoting the transplantability of the one or more biological cells after culture. In various embodiments of the microfluidic device, the at least one conditioning surface of the microfluidic device can comprise one or more components of mammalian serum. The one or more components of the mammalian serum may comprise B27® supplement, fetal bovine serum (FBS) or calf serum (FCS). In various embodiments of the microfluidic device, the microfluidic device may further comprise a substrate having a dielectrophoretic (DEP) configuration. In some embodiments, the substrate having a DEP configuration can be configured to introduce or remove one or more biological cells into the growth chamber. The DEP configuration can be actuated optically. In various embodiments of the microfluidic device, the at least one conditioning surface of the microfluidic device can be configured to be stable at a temperature of at least about 30°C. In various embodiments of the microfluidic device, the isolation region of the at least one growth chamber of the microfluidic device can have dimensions sufficient to support cell expansion to a range of about 100 cells. In some embodiments, no more than 1×10 ² biological cells can be maintained in the at least one growth chamber, and the volume of the at least one growth chamber can be less than or equal to about 2×10 ⁶ cubic microns. In other embodiments, no more than 1×10 ² biological cells can be maintained in the at least one growth chamber, and the volume of the at least one growth chamber can be less than or equal to about 1×10 ⁷ cubic microns. In various embodiments of the microfluidic device, the device can further comprise: at least one inlet configured to input the first fluidic medium or the second fluidic medium into the flow region; and at least one outlet, It is configured to receive the first fluid medium when the first fluid medium exits the flow region. In various embodiments of the microfluidic device, the microfluidic device can further comprise a deformable cover region over the at least one growth chamber or the isolation region thereof, whereby pressing the deformable cover region applies sufficient to cause the A force that biological cells output from the isolation region to the flow region. In various embodiments of the microfluidic device, the microfluidic device can include a lid, wherein at least a portion of the lid can be gas permeable, thereby providing a source of gas molecules to one positioned in the microfluidic device fluid medium. The gas permeable portion of the cover can be positioned over the at least one growth chamber. In other embodiments, the breathable portion of the cover may be positioned over the flow area. In other embodiments, the at least one growth chamber may include a plurality of growth chambers. In various embodiments, the one or more biological cells may comprise a plurality of biological cells. In various embodiments of the microfluidic device, the at least one growth chamber can comprise at least one surface conditioned to support cell growth, viability, transplantability, or any combination thereof of a mammalian cell. In other embodiments, the at least one growth chamber can comprise at least one surface conditioned to support cell growth, viability, transplantability, or any combination thereof of an immune cell. In other embodiments, the immune cell can be a lymphocyte or a white blood cell. In some other embodiments, the immune cell can be a B cell, a T cell, a NK cell, a macrophage or a dendritic cell. In various embodiments of the microfluidic device, the at least one growth chamber can comprise at least one surface conditioned to support cell growth, viability, transplantability, or any combination thereof of an attached cell. In various embodiments of the microfluidic device, the at least one growth chamber can comprise at least one surface conditioned to support cell growth, viability, transplantability, or any combination thereof of a fusion cell. In various embodiments of the microfluidic device, the at least one growth chamber may comprise cell growth, viability, portability, or any combination thereof, adjusted to support a single cell and a corresponding colony of biological cells at least one surface. In another aspect, a system for culturing one or more biological cells on a microfluidic device is provided, the system comprising a microfluidic device having: a flow region configured to contain a first a flow of a fluid medium; and at least one growth chamber, wherein the growth chamber has at least one surface conditioned to support cell growth, viability, portability, or any combination thereof in the microfluidic device. The at least one growth chamber may include an isolation region and a connecting region, the isolation region being fluidly connected to the connecting region and the connecting region having a proximal opening to the flow region. In some embodiments, the isolation region of the microfluidic device can be configured to contain a second fluid medium, and when the flow region and the at least one growth chamber are substantially filled with the first fluid medium and the second fluid medium, respectively In the case of a two-fluid medium, the components of the second fluid medium can diffuse into the first fluid medium and/or the components of the first fluid medium can diffuse into the second fluid medium, and the first medium can be substantially does not flow into this isolated area. In some embodiments, the microfluidic device can further comprise a microfluidic channel comprising at least a portion of the flow region, and wherein the connection region of the at least one growth chamber can directly communicate into the microfluidic channel. The microfluidic device can be any microfluidic device as described herein having any combination of any of the elements. In various embodiments of the system, the system can further comprise a flow controller configured to perfuse at least the first fluid medium. The controller is configured to discontinuously perfuse the at least first fluid medium. In various embodiments of the system, the microfluidic device of the system can further comprise a substrate having a dielectrophoretic (DEP) configuration configured to introduce one or more biological cells into the growth chamber or The one or more biological cells are removed from the growth chamber. The DEP configuration can be actuated optically. In various embodiments of the system, the system can further comprise a reservoir configured to contain the first fluid medium, wherein the reservoir is fluidly connected to the microfluidic device. The reservoir may be configured to communicate with a gaseous environment capable of saturating the first fluid medium with dissolved gas molecules. In various embodiments of the system, the system can further comprise a sensor connected to at least one inlet of the microfluidic device, wherein the sensor can be configured to detect a pH of the first fluid medium . In various embodiments of the system, the system can further comprise a sensor connected to at least one outlet, wherein the sensor is configured to detect the first fluidic medium as the first fluidic medium leaves the microfluidic device One pH of a fluid medium. In some embodiments, the sensor may be an optical sensor. In various embodiments of the system, the system can further include a detector configured to capture an image of the at least one growth chamber and any biological cells contained therein. In some embodiments, the one or more biological cells may comprise one or more mammalian cells. In other embodiments, the one or more biological cells may comprise one or more fusion tumor cells. In other embodiments, the one or more biological cells may comprise one or more lymphocytes or white blood cells. Alternatively, the one or more biological cells may comprise one or more adherent cells. In various embodiments of the system, the one or more biological cells in the growth chamber can be a single cell and the colony can be a colony of biological cells. In another aspect, there is provided a composition comprising: a substrate having a dielectrophoresis (DEP) configuration and a surface; and a conditioning surface covalently bonded to an oxide of the surface of the substrate part. The composition may have one of the structures of Formula 1 or Formula 2, and may have any value for the elements of Formula 1 or Formula 2, as defined herein:

Formula 1 Formula 2 In some embodiments of the composition, the conditioning surface may comprise a linking group covalently bonded to the oxide moieties of the surface, and the linking group may be bonded to a Part of being configured to support cell growth, viability, transplantability, or any combination thereof. In some embodiments, the linking group can be a siloxy linking group. In other embodiments, the linking group can be a phosphonate group. In some embodiments, the linking group can be directly linked to the moiety that is configured to support cell growth, viability, transplantability, or any combination thereof. In some embodiments, the linking group may be indirectly linked to the moiety that is configured to support cell growth, viability, transplantability, or any combination thereof. In some embodiments, the linking group may be indirectly linked to the moiety configured to support cell growth, viability, transplantability, or any combination thereof, via attachment to a linker. In some embodiments, the linking group may be indirectly linked via attachment to one of the first ends of a linker to a linker configured to support cell growth, viability, transplantability, or any combination thereof, etc. the section. The linker may further comprise a linear moiety, wherein a main chain of the linear moiety comprises 1 to 200 non-ferrous atoms selected from any combination of silicon atoms, carbon atoms, nitrogen atoms, oxygen atoms, sulfur atoms and phosphorus atoms A hydrogen atom. In some embodiments, the linker may further comprise a triazolyl moiety. In some embodiments, the triazolyl moiety can interrupt the linear portion of the linker or can be attached to a second end of the linear portion of the linker. In other embodiments, the backbone of the linear moiety may comprise an arylidene moiety. In various embodiments, the moiety configured to support cell growth, viability, transplantability, or any combination thereof, comprises an alkyl moiety, a fluoroalkyl moiety, a monosaccharide or polysaccharide, an ethanol moiety, a polyol moiety , alkylene ether moiety, polyelectrolyte moiety, amine moiety, carboxylic acid moiety, phosphonic acid moiety, sulfonate anion moiety, carboxybetaine moiety, sulfobetaine moiety, amine sulfonic acid moiety or amino acid moiety. In some embodiments, the at least one conditioning surface may comprise amino acids, alkyl moieties, perfluoroalkyl moieties, polydextrose moieties, and/or alkylene ether moieties. In some embodiments, the at least one conditioning surface may comprise alkyl or perfluoroalkyl moieties. In some embodiments, the alkyl or perfluoroalkyl moieties have a backbone length greater than one of 10 carbons. In various embodiments, the conditioning surface may further comprise one or more cleavable moieties. The lysable moiety can be configured to allow disruption of the regulatory surface, thereby promoting the transplantability of the one or more biological cells after culture. In another aspect, a method of culturing at least one biological cell in a microfluidic device having a flow region configured to contain a flow of a first fluid medium and at least one A growth chamber, the method comprising the steps of introducing the at least one biological cell into the at least one growth chamber, wherein the at least one growth chamber is configured to have modulation to support cell growth, viability, portability, or the like and at least one surface in any combination of the like; and incubating the at least one biological cell for a period of time at least sufficient to allow the at least one biological cell to expand to produce a colony of biological cells. The at least one growth chamber may include an isolation region and a connecting region, the isolation region being fluidly connected to the connecting region and the connecting region having a proximal opening to the flow region. In some embodiments, the isolation region of the microfluidic device can be configured to contain a second fluidic medium, and wherein the flow region and the at least one growth chamber are substantially filled with the first fluidic medium and the at least one growth chamber, respectively In the second fluid medium, components of the second fluid medium may diffuse into the first fluid medium and/or components of the first fluid medium may diffuse into the second fluid medium, and the first medium may There is substantially no flow into the isolation region. In some embodiments, the microfluidic device can further comprise a microfluidic channel having at least a portion of the flow region, and wherein the connecting region of the at least one growth chamber can directly communicate into the microfluidic channel. The microfluidic device can be any microfluidic device as described herein having any combination of any of the elements. In some embodiments of the method, the at least one conditioning surface can comprise a linking group covalently bonded to the surface, and wherein the linking group is further linked to a configuration to support the microfluidic device A portion of the cell growth, viability, transplantability, or any combination thereof, of the one or more biological cells within. In some other embodiments, the moiety configured to support cell growth, viability, transplantability, or any combination thereof, may comprise: an alkyl or fluoroalkyl (which includes perfluoroalkyl) moiety; a single moiety; Saccharides or polysaccharides (which may include, but are not limited to, polydextrose); alcohols (which include, but are not limited to, propargyl alcohol); polyols, which include, but are not limited to, polyvinyl alcohol; alkylene ethers, It includes, but is not limited to, polyethylene glycol; polyelectrolytes (which include, but are not limited to, polyacrylic acid or polyvinylphosphonic acid); amine groups (which include, but are not limited to, derivatives thereof, such as, but are not limited to, alkylated amines) , hydroxyalkylated amine groups, guanidine salts, and heterocyclic groups containing an unaromatic nitrogen ring atom, such as (but not limited to) morpholinyl or piperazinyl); carboxylic acids, including but not limited to propylene alkynoic acids (which can provide a carboxylate anion surface); phosphonic acids, including but not limited to ethynylphosphonic acids (which can provide a phosphonate anion surface); sulfonic acid anions; carboxybetaines; sulfobetaines; amines Sulfonic acid; or amino acid. In some embodiments, the at least one conditioning surface may comprise alkyl or perfluoroalkyl moieties. In other embodiments, the at least one conditioning surface may comprise alkylene ether moieties or polydextrose moieties. In some embodiments of the method, the method can include the step of conditioning at least one surface of the at least one growth chamber. In some embodiments, conditioning can include treating the at least one surface of the at least one growth chamber with a conditioning agent comprising a polymer. In other embodiments, conditioning may comprise treating at least one surface of the at least one growth chamber with one or more components of mammalian serum. In other embodiments, conditioning can include treating at least one surface of the at least one growth chamber with at least one cell adhesion blocking molecule. In some embodiments of the method, introducing the at least one biological cell into the at least one growth chamber can comprise: using a dielectrophoretic (DEP) force having a force sufficient to move the at least one biological cell. In some embodiments, using a DEP force may include optically actuating the DEP force. In some embodiments of the method, the method may further comprise the step of perfusing the first fluid medium during the incubation step, wherein the first fluid medium is introduced through at least one inlet of the microfluidic device and through the microfluidic device at least one outlet of the fluidic device is output, wherein after output, the first fluidic medium optionally includes components from the second fluidic medium. In some embodiments of the method, the method can further comprise the step of: after the incubation step, lysing one or more lysable moieties of the conditioning surface, thereby promoting the growth of the one or more biological cells from the The chamber or its isolated area is output into this flow area. In some embodiments of the method, the method may further comprise the step of exporting one or more biological cells from the growth chamber or its isolation region into the flow region. In some embodiments of the method, the at least one biological cell can comprise a mammalian cell. In other embodiments of the method, the at least one biological cell may comprise at least one immune cell. In other embodiments of the method, the at least one immune cell may comprise a lymphocyte or a white blood cell. In some other embodiments of the method, the at least one immune cell can comprise a B cell, a T cell, a NK cell, a macrophage or a dendritic cell. In other embodiments, the at least one biological cell may comprise an adherent cell. Alternatively, the at least one biological cell may comprise a fusion cell. In some embodiments of the method, the step of introducing the at least one biological cell into the at least one growth chamber may comprise introducing a single cell into the growth chamber, and the step of introducing the biological cell resulting from the incubation step The colony may be a selective colony. In another aspect, a kit for culturing a biological cell is provided, comprising a microfluidic device having: a flow region configured to flow a fluid containing a first fluid medium and at least one growth chamber comprising at least one surface conditioned to support cell growth, viability, transplantability, or any combination thereof within the microfluidic device. The at least one growth chamber may include an isolation region and a connecting region, the isolation region being fluidly connected to the connecting region and the connecting region having a proximal opening to the flow region. The microfluidic device can be any microfluidic device as described herein having any combination of elements. In some embodiments, the at least one conditioning surface of the microfluidic device may comprise alkyl moieties, fluoroalkyl moieties, monosaccharide or polysaccharide moieties, ethanol moieties, polyol moieties, alkylene ether moieties; polyelectrolyte moieties, An amine moiety, a carboxylic acid moiety, a phosphonic acid moiety, a sulfonate moiety, a carboxybetaine moiety, a sulfobetaine moiety, an amine sulfonic acid moiety, or an amino acid moiety. In some embodiments, the at least one conditioning surface of the microfluidic device includes at least one of: a carbohydrate moiety, an alkylene ether moiety, an alkyl moiety, a fluoroalkyl moiety, or an amino acid moiety. In some embodiments, the alkyl or fluoroalkyl moieties have a backbone length of greater than 10 carbons. In various embodiments of the kit, the at least one conditioning surface of the microfluidic device can comprise a linking group covalently bonded to a surface of the microfluidic device, and the linking group can be linked to a portion configured to support growth, viability, transplantability, or any combination thereof, of the one or more biological cells within the microfluidic device. In some embodiments, the linking group can be a siloxy linking group. In other embodiments, the linking group can be a phosphonate linking group. In some embodiments, the linking group can be directly linked to the moiety that is configured to support cell growth, viability, transplantability, or any combination thereof. In other embodiments, the linking group can be indirectly linked to the moiety that is configured to support cell growth, viability, transplantability, or any combination thereof. The linking group can be indirectly linked through a linker to the moiety configured to support cell growth, viability, transplantability, or any combination thereof. The linking group can be indirectly linked to the moiety configured to support cell growth, viability, transplantability, or any combination thereof, via attachment to a first end of a linker. In various embodiments, the linker may further comprise a linear moiety, wherein a backbone of the linear moiety includes a backbone selected from any combination of silicon atoms, carbon atoms, nitrogen atoms, oxygen atoms, sulfur atoms, and phosphorus atoms 1 to 200 non-hydrogen atoms. In some embodiments, the linker may further comprise a triazolyl moiety. In various embodiments of the kit, the kit can further comprise a surface conditioning agent. In some embodiments, the surface modifying agent may comprise a polymer comprising at least one of the following: an alkylene ether moiety, a carboxylic acid moiety, a sulfonic acid moiety, a phosphonic acid moiety, an amino acid moiety, a nucleic acid moiety, or sugar part. In some embodiments, the surface modifying agent may comprise a polymer comprising at least one of: an alkylene ether moiety, an amino acid moiety, and/or a carbohydrate moiety. In other embodiments, the surface modulating agent may comprise at least one cell adhesion blocking molecule. In some embodiments, the at least one cell adhesion blocking molecule disrupts actin fibril formation, blocks integrin receptors, or reduces cell binding to DNA-fouling surfaces. In some embodiments, the at least one cell adhesion blocking molecule can be cytochalasin B, an RGD-containing peptide, an inhibitor of fibronectin, an integrin antibody, or a DNase 1 protein. In some embodiments, the surface modulating agent may comprise a combination of one or more cell adhesion blocking molecules. In other embodiments, the surface modulating agent may comprise one or more components of mammalian serum. In some embodiments, the mammalian serum can be fetal bovine serum (FBS) or calf serum (FCS). In various embodiments of the kit, the kit can further comprise a medium supplement comprising an agent configured to enhance the conditioning of the at least one surface of the growth chamber. The medium supplement may contain a Pluronics® polymer. In various embodiments of the kit, the at least one conditioning surface of the microfluidic device can comprise a cleavable moiety. In some embodiments, the kit can further comprise an agent configured to cleave the cleavable portion of the modulating surface. In various embodiments of the kit, the kit can further comprise at least one reagent for detecting a state of the biological cell.

本申請案係根據35 U.S.C. 119(e)主張2015年4月22日申請之美國臨時申請案第62/151,325號(其之全部內容以引用的方式併入本文中)之權利的一非臨時申請案。微流體環境提供對一細胞或細胞群組提供一局部環境之機會，該局部環境依一時間相依方式及位置相依濃度將營養素及/或可溶細胞生長傳物種提供至該細胞或細胞群組。此等條件可表示更像活體內之生長條件或替代地容許擾動此等典型條件以容許非標準條件下之研究及生長。無法使用標準化大規模細胞培養方法來滿足此等要求。然而，需要改良方案來使一或若干細胞之操縱更容易以：a)將該(等)細胞放置至有益於支援細胞生長、存活性、可移植性或其等之任何組合之一微流體環境中；b)成功地維持該(等)細胞及/或擴增該(等)細胞之群體；及/或c)界定導致成功生長及/或維持之條件。本文中所描述之系統及方法允許用於微流體細胞培養之更精確細胞處置、環境控制及細胞隔離技術，且可用以產生(例如)選殖細胞群體。本說明書描述本發明之例示性實施例及應用。然而，本發明不受限於此等例示性實施例及應用或不受限於該等例示性實施例及應用之操作方式或其在本文中之描述方式。而且，圖式可展示簡化或局部視圖，且為清楚起見，圖式中之元件之尺寸可被放大或否則可不成比例。另外，當本文中使用術語「在...上」、「附接至」或「耦合至」時，一元件(例如一材料、一層、一基板等等)可「在另一元件上」、「附接至另一元件」或「耦合至另一元件」，無論該元件是否直接在該另一元件上、直接附接至該另一元件或直接耦合至該另一元件或在該元件與該另一元件之間存在一或多個介入元件。同樣地，方向(例如上方、下方、頂部、底部、側、向上、向下、下面、上面、上、下、水平、垂直、「x」、「y」、「z」等等)(若提供)係相對的，僅供例示的，用於使繪示及討論方便，且絕非意在限制。另外，當參考一元件系列(例如元件a、b、c)時，此參考意欲包含所列元件本身之任何者、非全部所列元件之任何組合及/或全部所列元件之一組合。本說明書中之章節劃分僅為了使檢查方便且不限制所討論之元件之任何組合。如本文中所使用，「實質上」意謂足以達成所欲目的。因此，術語「實質上」允許相對於一絕對或完美狀態、尺寸、量測、結果或其類似者之小幅、微不足道變動，諸如將由一般技術者預期且不會明顯地影響總體效能之變動。當與數值或參數或可表達為數值之特性一起使用時，「實質上」意謂在10%內。術語「若干」意謂一個以上。如本文中所使用，術語「複數個」可為2個、3個、4個、5個、6個、7個、8個、9個、10個或更多。如本文中所使用，「空氣」係指在地球之大氣中占主導地位之氣體之組合物。四種最豐富氣體係氮氣(通常存在約78體積%之一濃度，例如，在自約70%至約80%之一範圍內)、氧氣(在海平面處通常存在約20.95體積%，例如，在自約10%至約25%之一範圍內)、氬氣(通常存在約1.0體積%，例如，在自約0.1%至約3%之一範圍內)及二氧化碳(通常存在約0.04%，例如，在自約0.01%至約0.07%之一範圍內)。空氣可具有其他微量氣體，諸如甲烷、氧化亞氮或臭氧、微量污染物及有機材料，諸如花粉、柴油顆粒及其類似者。空氣可包含水蒸氣(通常存在約0.25%，或可存在自約10 ppm至約5體積%之一範圍)。空氣可經提供以在培養實驗中用作為一經過濾之受控組合物且可如本文中所描述被調節。如本文中所使用，術語「安置」涵蓋於其含義「定位」內。如本文中所使用，一「微流體器件」或「微流體裝置」係包含經構形以保持一流體之一或多個離散微流體線路之一器件，各微流體線路由流體地互連之線路元件(其包含(但不限於)(若干)區域、(若干)流動路徑、(若干)通道、(若干)室及/或(若干)圍欄)及至少兩個端口(其經構形以允許流體(及視情況，懸浮於流體中之微小物體)流動至微流體器件中及/或流出微流體器件)組成。通常，一微流體器件之一微流體線路將包含至少一微流體通道及至少一室，且將保持小於約1毫升(例如，小於約750微升、約500微升、約250微升、約200微升、約150微升、約100微升、約75微升、約50微升、約25微升、約20微升、約15微升、約10微升、約9微升、約8微升、約7微升、約6微升、約5微升、約4微升、約3微升或約2微升)之一流體體積。在某些實施例中，微流體線路保持約1微升至約2微升、約1微升至約3微升、約1微升至約4微升、約1微升至約5微升、約2微升至約5微升、約2微升至約8微升、約2微升至約10微升、約2微升至約12微升、約2微升至約15微升、約2微升至約20微升、約5微升至約20微升、約5微升至約30微升、約5微升至約40微升、約5微升至約50微升、約10微升至約50微升、約10微升至約75微升、約10微升至約100微升、約20微升至約100微升、約20微升至約150微升、約20微升至約200微升、約50微升至約200微升、約50微升至約250微升或約50微升至約300微升。如本文中所使用，一「奈流體器件」或「奈流體裝置」係具有一微流體線路之一類型之微流體器件，該微流體線路含有經構形以保持小於約1微升(例如，小於約750奈升、約500奈升、約250奈升、約200奈升、約150奈升、約100奈升、約75奈升、約50奈升、約25奈升、約20奈升、約15奈升、約10奈升、約9奈升、約8奈升、約7奈升、約6奈升、約5奈升、約4奈升、約3奈升、約2奈升、約1奈升或更小)之一流體體積之至少一線路元件。通常，一奈流體器件將包括複數個線路元件(例如至少2個、3個、4個、5個、6個、7個、8個、9個、10個、15個、20個、25個、50個、75個、100個、150個、200個、250個、300個、400個、500個、600個、700個、800個、900個、1000個、1500個、2000個、2500個、3000個、3500個、4000個、4500個、5000個、6000個、7000個、8000個、9000個、10000個或更多)。在某些實施例中，該至少一線路元件之一或多者(例如全部)經構形以保持約100皮升至約1奈升、約100皮升至約2奈升、約100皮升至約5奈升、約250皮升至約2奈升、約250皮升至約5奈升、約250皮升至約10奈升、約500皮升至約5奈升、約500皮升至約10奈升、約500皮升至約15奈升、約750皮升至約10奈升、約750皮升至約15奈升、約750皮升至約20奈升、約1奈升至約10奈升、約1奈升至約15奈升、約1奈升至約20奈升、約1奈升至約25奈升或約1奈升至約50奈升之一流體體積。在其他實施例中，該至少一線路元件之一或多者(例如全部)經構形以保持約100奈升至約200奈升、約100奈升至約300奈升、約100奈升至約400奈升、約100奈升至約500奈升、約200奈升至約300奈升、約200奈升至約400奈升、約200奈升至約500奈升、約200奈升至約600奈升、約200奈升至約700奈升、約250奈升至約400奈升、約250奈升至約500奈升、約250奈升至約600奈升或約250奈升至約750奈升之一流體體積。如本文中所使用，一「微流體通道」或「流動通道」係指一微流體器件之流動區域，其具有顯著地長於水平尺寸及垂直尺寸兩者之一長度。例如，流動通道可為水平尺寸或垂直尺寸之至少5倍長度，例如至少10倍長度、至少25倍長度、至少100倍長度、至少200倍長度、至少300倍長度、至少400倍長度、至少500倍長度或更長。在一些實施例中，一流動通道之長度係在自約20,000微米至約100,000微米之範圍內，其包含其間之任何範圍。在一些實施例中，水平尺寸係在自約100微米至約1000微米(例如約150微米至約500微米)之範圍內且垂直尺寸係在自約25微米至約200微米(例如自約40微米至約150微米)之範圍內。應注意，一流動通道可在一微流體器件中具有各種不同空間構形，且因此不受限於一完全線性元件。例如，一流動通道可為或包含具有下列構形之一或多個區段：曲線、彎曲、螺旋、傾斜、下傾、分叉(例如多個不同流動路徑)及其等之任何組合。另外，一流動通道可具有沿其路徑之不同加寬及收縮橫截面積以在其內提供一所要流體流動。如本文中所使用，術語「障礙」一般係指一凸塊或類似類型之結構，其足夠大以便部分(但非完全)阻礙目標微小物體在一微流體器件中之兩個不同區域或線路元件之間的移動。該兩個不同區域/線路元件可為(例如)一微流體培育室及一微流體通道或一微流體培育室之一連接區域及一隔離區域。如本文中所使用，術語「收縮」一般係指一微流體器件中之一線路元件(或兩個線路元件之間的一界面)之一寬度之一變窄。收縮可定位於(例如)一微流體培育室與一微流體通道之間的界面處或定位於一微流體培育室之一隔離區域與一連接區域之間的界面處。如本文中所使用，「透明」係指允許可見光穿過且實質上不會在該光穿過時更改該光之一材料。如本文中所使用，術語「微小物體」一般係指可根據本發明而隔離及收集之任何微觀物體。微小物體之非限制性實例包含：無生命微小物體，諸如微粒；微珠(例如聚苯乙烯珠粒、Luminex™珠粒或其類似者)；磁珠；微桿；微絲；量子點及其類似者；生物微小物體，諸如細胞(例如胚胎細胞、卵母細胞、精子細胞、自一組織解離之細胞、真核細胞、原生生物細胞、動物細胞、哺乳動物細胞、人類細胞、免疫細胞(其包含(但不限於) T細胞、B細胞、自然殺手細胞、巨噬細胞、樹突細胞及其類似者)、融合瘤細胞、培養細胞、來自一細胞系之細胞、癌細胞(其包含(但不限於)循環腫瘤細胞)、受感染細胞、轉染及/或轉形細胞(其包含(但不限於) CHO細胞)、報導體細胞、原核細胞及其類似者)；生物細胞器(例如細胞核)；囊泡或複合物；合成囊泡；脂質體(例如，合成的或衍生自膜配製物)；脂質奈筏(如Ritchie等人(2009)之「Reconstitution of Membrane Proteins in Phospholipid Bilayer Nanodiscs」(Methods Enzymol.，464:211-231)中所描述)及其類似者；或無生命微小物體及生物微小物體之一組合(例如附接至細胞之微珠、經脂質體塗佈之微珠、經脂質體塗佈之磁珠或其類似者)。珠粒可進一步具有共價或非共價鍵聯之其他部分/分子，諸如螢光標記、蛋白質、小分子傳信部分、抗原或能夠用於一化驗中之化學/生物物種。如本文中所使用，術語「細胞」係指一生物細胞，其可為一植物細胞、一動物細胞(例如一哺乳動物細胞)、一細菌細胞、一真菌細胞或其類似者。一哺乳動物細胞可來自(例如)一人、一小鼠、一大鼠、一馬、一山羊、一綿羊、一牛、一靈長類動物或其類似者。若能夠生殖之一生物細胞群落中之全部活細胞係衍生自一單一母細胞之子細胞，則該群落係「選殖的」。術語「選殖細胞」係指相同選殖群落之細胞。如本文中所使用，生物細胞之「群落」係指2個或2個以上細胞(例如2個至20個、4個至40個、6個至60個、8個至80個、10個至100個、20個至200個、40個至400個、60個至600個、80個至800個、100個至1000個或大於1000個細胞)。如本文中所使用，術語「維持(一或若干)細胞」係指提供包括流體組分及氣體組分兩者(其等提供使細胞保持存活及/或擴增所需之條件)之一環境。如本文中所使用，術語「擴增」在涉及細胞時係指細胞數目之增加。如本文中所涉及，「可透氣」意謂材料或結構可滲透氧氣、二氧化碳或氮氣之至少一者。在一些實施例中，可透氣材料或結構可滲透氧氣、二氧化碳及氮氣之一者以上且可進一步滲透此等氣體之全部三者。一流體培養基之一「組分」係存在於該培養基中之任何化學或生化分子，其包含溶劑分子、離子、小分子、抗生素、核苷酸及核苷、核酸、胺基酸、肽、蛋白質、糖、碳水化合物、脂質、脂肪酸、膽固醇、代謝物或其類似者。如本文中參考一流體培養基所使用，「擴散」係指該流體培養基之一組分依一濃度梯度之熱力移動。片語「一培養基之流動」意謂主要歸因於除擴散之外之任何機制之一流體培養基之整體移動。例如，一培養基之流動可涉及該流體培養基自一點至另一點之移動(歸因於該等點之間的壓力差)。此流動可包含液體之一連續、脈衝、週期性、隨機、間歇或往復流動或其等之任何組合。當一流體培養基流動至另一流體培養基中時，可導致該等培養基之紊流及混合。片語「實質上無流動」係指一流體培養基之一流動速率，其當隨時間平均化時小於一材料(例如所關注之一分析物)之組分擴散至該流體培養基中或該流體培養基內之速率。此一材料之組分之擴散速率可取決於(例如)溫度、該等組分之大小及該等組分與該流體培養基之間的相互作用之強度。如本文中參考一微流體器件內之不同區域所使用，片語「流體地連接」意謂：當該等不同區域實質上填充有流體(諸如流體培養基)時，該等區域之各者中之流體經連接以便形成一單一流體。此不意謂該等不同區域中之流體(或流體培養基)需要具有相同組合物。確切而言，一微流體器件之不同流體連接區域中之流體可具有不同組合物(例如不同濃度之溶質，諸如蛋白質、碳水化合物、離子或其他分子)，其等隨著溶質依其各自濃度梯度移動及/或流體流動通過該器件而變化。一微流體(或奈流體)器件可包括「掃及」區域及「未掃及」區域。如本文中所使用，一「掃及」區域由一微流體線路之一或多個流體互連線路元件組成，該一或多個流體互連線路元件之各者在流體流動通過該微流體線路時經歷一培養基之流動。一掃及區域之該等線路元件可包含(例如)區域、通道及全部或部分室。如本文中所使用，一「未掃及」區域由一微流體線路之一或多個流體互連線路元件組成，該一或多個流體互連線路元件之各者在流體流動通過該微流體線路時實質上未經歷流體通量。一未掃及區域可流體地連接至一掃及區域，前提是：該等流體連接經結構化以實現培養基在該掃及區域與該未掃及區域之間的擴散但實質上未實現培養基在該掃及區域與該未掃及區域之間的流動。因此，該微流體器件可經結構化以實質上使一未掃及區域與一掃及區域中之培養基之一流動隔離，同時實質上僅實現該掃及區域與該未掃及區域之間的擴散性流體連通。例如，一微流體器件之一流動通道係一掃及區域之一實例，而一微流體器件之一隔離區域(下文將進一步詳細描述)係一未掃及區域之一實例。如本文中所使用，流體培養基流之一「非掃及」速率意謂足以容許生長室之一隔離區域中之一第二流體培養基之組分擴散至流動區域中之第一流體培養基中及/或容許第一流體培養基之組分擴散至隔離區域中之第二流體培養基中之一流動速率；且其中第一培養基實質上未進一步流動至隔離區域中。如本文中所使用，一「流動路徑」係指一或多個流體連接線路元件(例如(若干)通道、(若干)區域、(若干)室及其類似者)，其等界定培養基之一流動之軌跡且經受培養基之一流動。因此，一流動路徑係一微流體器件之一掃及區域之一實例。其他線路元件(例如未掃及區域)可與包括該流動路徑但未經受該流動路徑之培養基之流動的線路元件流體地連接。如本文中所使用，「伸芳基」係指具有6個至10個環原子之芳香族基(例如，C6-C10芳香族或C6-C10芳基)，其具有至少一環(其具有呈碳環形之一共軛π電子系統(例如苯基、茀基及萘基))且具有附接至一分子之其他部分之一或兩個點。每當一數值範圍(諸如「6至10」)出現於本文中時，其係指該給定範圍內之各整數；例如，「6個至10個環原子」意謂芳基可由6個環原子、7個環原子等等(直至且包含10個環原子)組成。該術語包含單環或稠環多環(即，共用環原子之相鄰對之環)基。伸芳基之實例包含(但不限於)伸苯基、伸萘基及其類似者)。伸芳基部分可進一步經取代或可僅具有附接至分子之其他部分之一或兩點之取代。如本文中所使用，「伸雜芳基」係指一5員至18員芳香族基(例如C3-C13雜芳基)，其包含選自氮、氧及硫之一或多個環雜原子，且可包含一單環、雙環、三環或四環系統且前綴「伸」指示雜芳基環系統具有附接至一分子之其他部分之一或兩個點。每當一數值範圍(諸如「5至18」)出現於本文中時，其係指該給定範圍內之各整數；例如，「5個至18個環原子」意謂雜芳基可由5個環原子、6個環原子等等(直至且包含18個環原子)組成。一含N之「雜芳香族」或「雜芳基」部分係指芳香基，其中環之骨骼原子之至少一者係一氮原子。多環雜芳基可為稠合的或非稠合的。可視情況氧化雜芳基中之(若干)雜原子。可視情況季銨化一或多個氮原子(若存在)。雜芳基透過(若干)環之任何原子而附接至分子之剩餘部分。伸雜芳基之實例包含(但不限於)伸苯并咪唑基、伸苯吲哚基、伸異噁唑基、伸噻唑基、伸三唑基、伸四唑基及伸苯硫基(即，伸噻吩基)。伸雜芳基部分可進一步經取代或可僅具有附接至該分子之其他部分之一或兩個點之取代。如本文中所使用，術語「雜環」係指：一經或未經3員、4員、5員、6員或7員取代之飽和或部分不飽和環，其含有1個、2個或3個雜原子，較佳地，含有獨立地選自氧、氮及硫之1個或2個雜原子；或一雙環系統，其含有高達10個原子，該等原子包含獨立地選自氧、氮及硫之至少一雜原子，其中含有雜原子之環係飽和的。雜環基之實例包含(但不限於)四氫呋喃基、四氫糠基、吡咯啶基、六氫吡啶基、4-吡喃基、四氫吡喃基、硫㖦基、嗎啉基、六氫吡嗪基、二氧戊環基、二氧己環基、吲哚啉基及5-甲基-6-苯并二氫吡喃基。雜環基具有附接至分子之其他部分之一或兩個點或可進一步經取代或可不進一步經取代。系統本發明提供一種用於在一微流體器件中培養一或多個生物細胞之系統，其包含一微流體器件，該微流體器件包括：一流動區域，其經構形以含有一第一流體培養基之一流動；及至少一生長室，其中該生長室具有經調節以支援細胞生長、存活性、可移植性或其等之任何組合之至少一表面。 微流體器件及用於操作 及 觀測此等器件之系統圖1繪示可用於本發明之實踐中之一微流體器件100及一系統150之一實例。圖中展示微流體器件100之一透視圖，其使其蓋110經部分切除以提供微流體器件100之一部分視圖。微流體器件100大體上包括一微流體線路120，其包括一流動路徑106，一流體培養基180可流動通過流動路徑106以視情況運送一或多個微小物體(圖中未展示)至微流體線路120中及/或通過微流體線路120。雖然圖1中繪示一單一微流體線路120，但適合微流體器件可包含複數個(例如2個或3個)此等微流體線路。無論如何，微流體器件100可經構形為一奈流體器件。在圖1所繪示之實施例中，微流體線路120包括複數個微流體生長室124、126、128及130，其等各具有與流動路徑106流體連通之一或多個開口。如下文將進一步討論，微流體生長室包括各種特徵及結構，其等已經最佳化以即使在一培養基180流動通過流動路徑106時亦使微小物體保留於微流體器件(諸如微流體器件100)中。然而，在開始描述上述各者之前，提供微流體器件100及系統150之一簡要描述。如圖1中大體上所繪示，微流體線路120由一封閉體102界定。雖然可將封閉體102實體地結構化成不同構形，但在圖1所展示之實例中，將封閉體102描繪為包括一支撐結構104 (例如一基底)、一微流體線路結構108及一蓋110。支撐結構104、微流體線路結構108及蓋110可彼此附接。例如，微流體線路結構108可安置於支撐結構104之一內表面109上，且蓋110可安置於微流體線路結構108上方。微流體線路結構108可與支撐結構104及蓋110一起界定微流體線路120之元件。支撐結構104可位於微流體線路120之底部處且蓋110可位於微流體線路120之頂部處，如圖1中所繪示。替代地，可使支撐結構104及蓋110依其他定向構形。例如，支撐結構104可位於微流體線路120之頂部處且蓋110可位於微流體線路120之底部處。無論如何，可存在各包括進入或離開封閉體102之一通路之一或多個端口107。一通路之實例包含一閥、一閘、一貫通孔或其類似者。如圖中所繪示，端口107係由微流體線路結構108中之一間隙產生之一貫通孔。然而，端口107可位於封閉體102之其他組件(諸如蓋110)中。雖然圖1中僅繪示一個端口107，但微流體線路120可具有兩個或兩個以上端口107。例如，可存在用作使流體進入微流體線路120之一入口之一第一端口107，且可存在用作使流體退出微流體線路120之一出口之一第二端口107。一端口107是否用作一入口或一出口可取決於流體流動通過流體路徑106之方向。支撐結構104可包括一或多個電極(圖中未展示)及一基板或複數個互連基板。例如，支撐結構104可包括一或多個半導體基板，其等之各者電連接至一電極(例如，該等半導體基板之全部或一子集可電連接至一單一電極)。支撐結構104可進一步包括一印刷電路板總成(「PCBA」)。例如，該(等)半導體基板可安裝於一PCBA上。微流體線路結構108可界定微流體線路120之線路元件。此等線路元件可包括可在微流體線路120填充有流體時流體地互連之空間或區域，諸如流動通道、室、圍欄、捕集器及其類似者。在圖1所繪示之微流體線路120中，微流體線路結構108包括一框架114及一微流體線路材料116。框架114可部分或完全圍封微流體線路材料116。框架114可為(例如)實質上包圍微流體線路材料116之一相對剛性結構。例如，框架114可包括一金屬材料。可使用空穴或其類似者來圖案化微流體線路材料116以界定微流體線路120之線路元件及互連件。微流體線路材料116可包括可透氣之一可撓性材料，諸如一可撓性聚合物(例如橡膠、塑膠、彈性體、聚矽氧、聚二甲矽氧烷(「PDMS」)或其類似者)。可構成微流體線路材料116之材料之其他實例包含模製玻璃、一可蝕刻材料(諸如聚矽氧(例如光可圖案化聚矽氧或「PPS」)、光阻劑(例如SU8)或其類似者。在一些實施例中，此等材料(及因此微流體線路材料116)可為剛性的及/或實質上可不透氣的。無論如何，微流體線路材料116可安置於支撐結構104上及框架114內。蓋110可為框架114及/或微流體線路材料116之一整合部分。替代地，蓋110可為一不同結構元件，如圖1中所繪示。蓋110可包括相同於或不同於框架114及/或微流體線路材料116之材料。類似地，支撐結構104可為與框架114或微流體線路材料116分離之一結構(如圖中所繪示)或為框架114或微流體線路材料116之一整合部分。同樣地，框架114及微流體線路材料116可為分離結構(如圖1中所展示)或為相同結構之整合部分。在一些實施例中，蓋110可包括一剛性材料。該剛性材料可為玻璃或具有類似性質之一材料。在一些實施例中，蓋110可包括一可變形材料。該可變形材料可為一聚合物，諸如PDMS。在一些實施例中，蓋110可包括剛性材料及可變形材料兩者。例如，蓋110之一或多個部分(例如定位於生長室124、126、128、130上方之一或多個部分)可包括與蓋110之剛性材料界接之一可變形材料。在一些實施例中，蓋110可進一步包含一或多個電極。該一或多個電極可包括可塗佈於玻璃或一類似絕緣材料上之一導電氧化物，諸如氧化銦錫(ITO)。替代地，該一或多個電極可為嵌入於一可變形材料(諸如一聚合物(例如PDMS))中之可撓性電極，諸如單壁奈米管、多壁奈米管、奈米線、導電奈米粒子之叢集或其等之組合。例如，U.S. 2012/0325665 (Chiou等人)中已描述可用於微流體器件中之可撓性電極，該案之內容以引用的方式併入本文中。在一些實施例中，蓋110可經改質(例如，藉由調節向內面向微流體線路120之一表面之全部或部分)以支援細胞黏著、存活性及/或生長。該改質可包含塗佈一合成或天然聚合物。在一些實施例中，蓋110及/或支撐結構104可為透光的。蓋110亦可包含至少一可透氣材料，例如PDMS或PPS。圖1亦展示用於操作及控制微流體器件(諸如微流體器件100)之一系統150。如圖中所繪示，系統150包含一電源192、一成像器件194及一傾斜器件190。電源192可將電力提供至微流體器件100及/或傾斜器件190以根據需要提供偏壓電壓或電流。電源192可(例如)包括一或多個交流(AC)及/或直流(DC)電壓源或電流源。成像器件194可包括用於擷取微流體線路120內之影像之一器件，諸如一數位攝影機。在一些例項中，成像器件194進一步包括具有一快圖框速率及/或高敏感度(例如，用於弱光應用)之一偵測器。成像器件194亦可包含用於將刺激輻射及/或光束導引至微流體線路120中且收集自微流體線路120 (或含於其內之微小物體)反射或發射之輻射及/或光束之一機構。發射光束可在可見光譜中且可(例如)包含螢光發射。反射光束可包含源自一LED或一寬光譜燈(諸如一汞燈(例如一高壓汞燈)或一氙弧燈)之反射發射。如關於圖3所討論，成像器件194可進一步包含一顯微鏡(或一光學元件串)，其可或可不包含一目鏡。系統150可進一步包括經構形以使一微流體器件100圍繞一或多個旋轉軸旋轉之一傾斜器件190。在一些實施例中，傾斜器件190經構形以圍繞至少一軸支撐及/或保持包括微流體線路120之封閉體102，使得微流體器件100 (及因此微流體線路120)可保持於一水平定向(即，相對於x軸及y軸成0°)、一垂直定向(即，相對於x軸及/或y軸成90°)或其間之任何定向上。微流體器件100 (及微流體線路120)相對於一軸之定向在本文中指稱微流體器件100 (及微流體線路120)之「傾角」。例如，傾斜器件190可使微流體器件100相對於x軸傾斜0.1°、0.2°、0.3°、0.4°、0.5°、0.6°、0.7°、0.8°、0.9°、1°、2°、3°、4°、5°、10°、15°、20°、25°、30°、35°、40°、45°、50°、55°、60°、65°、70°、75°、80°、90°或其間之任何度數。水平定向(及因此x軸及y軸)經界定為法向於由重力界定之一垂直軸。傾斜器件亦可使微流體器件100 (及微流體線路120)相對於x軸及/或y軸傾斜大於90°之任何度數，或使微流體器件100 (及微流體線路120)相對於x軸或y軸傾斜180°以使微流體器件100 (及微流體線路120)完全倒置。類似地，在一些實施例中，傾斜器件190使微流體器件100 (及微流體線路120)圍繞由微流體線路120之流動路徑106或某一其他部分界定之一旋轉軸傾斜。在一些例項中，使微流體器件100傾斜成一垂直定向，使得流動路徑106定位於一或多個生長室上方或一或多個生長室下方。如本文中所使用，術語「上方」指示：在由重力界定之一垂直軸上，流動路徑106定位成高於一或多個生長室(即，一流動路徑106上方之一生長室中之一物體將具有比流動路徑中之一物體高之一重力勢能)。如本文中所使用，術語「下方」指示：在由重力界定之一垂直軸上，流動路徑106定位成低於一或多個生長室(即，一流動路徑106下方之一生長室中之一物體將具有比流動路徑中之一物體低之一重力勢能)。在一些例項中，傾斜器件190使微流體器件100圍繞平行於流動路徑106之一軸傾斜。而且，可使微流體器件100傾斜小於90°之一角度，使得流動路徑106定位於一或多個生長室上方或一或多個生長室下方且非直接定位於生長室上方或生長室下方。在其他例項中，傾斜器件190使微流體器件100圍繞垂直於流動路徑106之一軸傾斜。在其他例項中，傾斜器件190使微流體器件100圍繞既不平行於流動路徑106且不垂直於流動路徑106之一軸傾斜。系統150可進一步包含一培養基源178。培養基源178 (例如一容器、儲液器或其類似者)可包括各用於保持一不同流體培養基180之多個區段或容器。因此，培養基源178可為位於微流體器件100外且與微流體器件100分離之一器件，如圖1中所繪示。替代地，培養基源178可全部或部分定位於微流體器件100之封閉體102內。例如，培養基源178可包括儲液器，其係微流體器件100之部分。圖1亦繪示控制及監測設備152之實例之簡化方塊圖描述，控制及監測設備152構成系統150之部分且可與一微流體器件100一起使用。如圖中所展示，此控制及監測設備152之實例包含：一主控制器154，其包括：用於控制培養基源178之一培養基模組160、用於控制微流體線路120中之微小物體(圖中未展示)及/或培養基(例如培養基之液滴)之移動及/或選擇之一動力模組162、用於控制一成像器件194 (例如一攝影機、顯微鏡、光源或其等之任何組合)來擷取影像(例如數位影像)之一成像模組164、及用於控制一傾斜器件190之一傾斜模組166。控制設備152亦可包含用於控制、監測或執行關於微流體器件100之其他功能之其他模組168。如圖中所展示，設備152可進一步包含一顯示器件170及一輸入/輸出器件172。主控制器154可包括一控制模組156及一數位記憶體158。控制模組156可包括(例如)一數位處理器，其經構形以根據作為非暫時性資料或信號儲存於記憶體158中之機械可執行指令(例如軟體、韌體、原始程式碼或其類似者)而操作。替代地或另外，控制模組156可包括固線式數位電路及/或類比電路。可類似地構形培養基模組160、動力模組162、成像模組164、傾斜模組166及/或其他模組168。因此，可由如上文所討論般構形之主控制器154、培養基模組160、動力模組162、成像模組164、傾斜模組166及/或其他模組168之任何一或多者執行如關於微流體器件100或任何其他微流體裝置而執行之本文中所討論之功能、程序、行動、動作或一程序之步驟。類似地，主控制器154、培養基模組160、動力模組162、成像模組164、傾斜模組166及/或其他模組168可經通信地耦合以傳輸及接收用於本文中所討論之任何功能、程序、行動、動作或步驟中之資料。培養基模組160控制培養基源178。例如，培養基模組160可控制培養基源178將一選定流體培養基180輸入至封閉體102中(例如，透過一入口107)。培養基模組160亦可控制自封閉體102移除培養基(例如，透過一出口(圖中未展示))。因此，可將一或多個培養基選擇性地輸入至微流體線路120中及自微流體線路120移除。培養基模組160亦可控制流體培養基180在微流體線路120內之流動路徑106中之流動。例如，在一些實施例中，在傾斜模組166致使傾斜器件190使微流體器件100傾斜一所要傾斜角之前，培養基模組160停止培養基180在流動路徑106中流動且停止培養基180流動通過封閉體102。動力模組162可經構形以控制微流體線路120中之微小物體(圖中未展示)之選擇、捕集及移動。如下文將關於圖2A及圖2B而討論，封閉體102可包括一介電泳(DEP)、光電鑷子(OET)及/或光電潤濕(OEW)構形(圖1中未展示)，且動力模組162可控制啟動電極及/或電晶體(例如光電晶體)來選擇及移動流動路徑106及/或生長室124、126、128、130中之微小物體(圖中未展示)及/或培養基之液滴(圖中未展示)。成像模組164可控制成像器件194。例如，成像模組164可接收及處理來自成像器件194之影像資料。來自成像器件194之影像資料可包括由成像器件194擷取之任何類型之資訊(例如，存在或不存在微小物體、培養基之液滴、標記(諸如螢光標記)之累積等等)。成像模組164可使用由成像器件194擷取之資訊來進一步計算物體(例如微小物體、培養基之液滴)之位置及/或此等物體在微流體器件100內之運動速率。傾斜模組166可控制傾斜器件190之傾斜運動。替代地或另外，傾斜模組166可控制傾斜速率及時序以最佳化微小物體經由重力而轉移至一或多個生長室。傾斜模組166與成像模組164通信地耦合以接收描述微小物體及/或培養基之液滴在微流體線路120中之運動之資料。傾斜模組166可使用此資料來調整微流體線路120之傾角以調整微小物體及/或培養基之液滴在微流體線路120中之移動速率。傾斜模組166亦可使用此資料來反覆地調整一微小物體及/或培養基之液滴在微流體線路120中之位置。在圖1所展示之實例中，將微流體線路120繪示為包括一微流體通道122及生長室124、126、128、130。各室包括至通道122之一開口，但其他部分經封閉使得室可實質上使室內之微小物體與通道122之流動路徑106中或其他室中之流體培養基180及/或微小物體隔離。在一些實施例中，室124、126、128、130經構形以實體地圈住微流體線路120內之一或多個微小物體。根據本發明之生長室可包括各種形狀、表面及特徵，其等經最佳化以與DEP、OET、OEW及/或重力一起使用，如將在下文詳細討論及展示。微流體線路120可包括任何數目個微流體生長室。雖然圖中展示5個生長室，但微流體線路120可具有更少或更多生長室。在一些實施例中，微流體線路120包括複數個微流體生長室，其中該等生長室之兩者或兩者以上包括不同結構及/或特徵。在圖1所繪示之實施例中，展示一單一通道122及流動路徑106。然而，其他實施例可含有各經構形以包括一流動路徑106之多個通道122。微流體線路120進一步包括與流動路徑106及流體培養基180流體連通之一入口閥或入口107，藉此流體培養基180可經由入口107而進入通道122。在一些例項中，流動路徑106包括一單一路徑。在一些例項中，將該單一路徑配置成一Z字形圖案，藉此流動路徑106在交替方向上橫跨微流體器件100行進兩次或兩次以上。在一些例項中，微流體線路120包括複數個並行通道122及流動路徑106，其中各流動路徑106內之流體培養基180在相同方向上流動。在一些例項中，各流動路徑106內之流體培養基在一向前方向或一相反方向之至少一者上流動。在一些例項中，複數個生長室經構形(例如，相對於一通道122)使得其等可並行地裝載有目標微小物體。在一些實施例中，微流體線路120進一步包括一或多個微小物體捕集器132。捕集器132一般形成於形成一通道122之邊界之一壁中，且可經定位成相對於微流體生長室124、126、128、130之一或多者之一開口。在一些實施例中，捕集器132經構形以接收或捕獲來自流動路徑106之一單一微小物體。在一些實施例中，捕集器132經構形以接收或捕獲來自流動路徑106之複數個微小物體。在一些例項中，捕集器132包括近似等於一單一目標微小物體之體積的一體積。捕集器132可進一步包括經構形以有助於目標微小物體流動至捕集器132中之一開口。在一些例項中，捕集器132包括具有近似等於一單一目標微小物體之尺寸之一高度及寬度之一開口，藉此防止較大微小物體進入至微小物體捕集器中。捕集器132可進一步包括經構形以有助於目標微小物體保留於捕集器132內之其他特徵。在一些例項中，捕集器132與一微流體生長室之開口對準且位於與一微流體生長室之開口相對之一通道122之側上，使得在使微流體器件100圍繞平行於通道122之一軸傾斜之後，所捕集之微小物體依致使微小物體降落至生長室之開口中之一軌跡退出捕集器132。在一些例項中，捕集器132包括一側通路134，其小於目標微小物體以促進流動通過捕集器132且藉此增加捕獲捕集器132中之一微小物體之可能性。在一些實施例中，經由一或多個電極(圖中未展示)而橫跨流體培養基180 (例如，在流動路徑中及/或在生長室中)施加介電泳(DEP)力以操縱、運輸、分離及分類定位於流動路徑及/或生長室中之微小物體。例如，在一些實施例中，將DEP力施加於微流體線路120之一或多個部分以將一單一微小物體自流動路徑106轉移至一所要微流體生長室中。在一些實施例中，DEP力用以防止一生長室(例如生長室124、126、128或130)內之一微小物體自該生長室排出。進一步言之，在一些實施例中，DEP力用以自一生長室選擇性地移除先前根據本發明之教示而收集之一微小物體。在一些實施例中，DEP力包括光電鑷子(OET)力。在其他實施例中，經由一或多個電極(圖中未展示)而將光電潤濕(OEW)力施加於微流體器件100之支撐結構104 (及/或蓋110)中之一或多個位置(例如有助於界定流動路徑及/或生長室之位置)以操縱、運輸、分離及分類定位於微流體線路120中之液滴。例如，在一些實施例中，將OEW力施加於支撐架構104 (及/或蓋110)中之一或多個位置以將一單一液滴自流動路徑106轉移至一所要微流體生長室中。在一些實施例中，OEW力用以防止一生長室(例如生長室124、126、128或130)內之一液滴自該生長室排出。進一步言之，在一些實施例中，OEW力用以自一生長室選擇性地移除先前根據本發明之教示而收集之一液滴。在一些實施例中，DEP力及/或OEW力與其他力(諸如流動力及/或重力)組合以便操縱、運輸、分離及分類微流體線路120內之微小物體及/或液滴。例如，封閉體102可經傾斜(例如，藉由傾斜器件190)以將流動路徑106及定位於流動路徑106內之微小物體定位於微流體生長室上方，且重力可將微小物體及/或液滴運輸至室中。在一些實施例中，可在其他力之前施加DEP力及/或OEW力。在其他實施例中，可在其他力之後施加DEP力及/或OEW力。在其他例項中，DEP力及/或OEW力可與其他力同時被施加或與其他力依一交替方式被施加。圖2A至圖2F繪示可用於本發明之實踐中之微流體器件之各種實施例。圖2A描繪其中微流體器件200經構形為一光學致動之電動器件之一實施例。此項技術中已知各種光學致動之電動器件，其包含具有一光電鑷子(OET)構形之器件及具有一光電潤濕(OEW)構形之器件。下列美國專利文件(其等之各者之全文以引用的方式併入本文中)中繪示適合OET構形之實例：美國專利第RE 44,711號(Wu等人)(最初發表為美國專利第7,612,355號)及美國專利第7,956,339號(Ohta等人)。美國專利第6,958,132號(Chiou等人)及美國專利申請公開案第2012/0024708號(Chiou等人)中繪示OEW構形之實例，該兩個專利之全文以引用的方式併入本文中。一光學致動之電動器件之又一實例包含一組合之OET/OEW構形，美國專利公開案第20150306598號(Khandros等人)及第20150306599號(Khandros等人)及其對應PCT公開案WO2015/164846及WO2015/164847中展示該組合之OET/OEW構形之實例，全部該等案之全文以引用的方式併入本文中。 動力微流體器件構形如上文所描述，系統之控制及監測設備可包括用於選擇及移動一微流體器件之微流體線路中之物體(諸如微小物體或液滴)之一動力模組。微流體器件可具有取決於所移動之物體之類型及其他考量之各種動力構形。例如，可利用一介電泳(DEP)構形來選擇及移動微流體線路中之微小物體。因此，微流體器件100之支撐結構104及/或蓋110可包括用於選擇性地誘發微流體線路120中之一流體培養基180中之微小物體上之DEP力之一DEP構形且藉此選擇、捕獲及/或移動個別微小物體或微小物體群組。替代地，微流體器件100之支撐結構104及/或蓋110可包括用於選擇性地誘發微流體線路120中之一流體培養基180中之液滴上之電潤濕(EW)力之一EW構形且藉此選擇、捕獲及/或移動個別液滴或液滴群組。圖2A及圖2B中繪示包括一DEP構形之一微流體器件200之一實例。雖然為簡單起見，圖2A及圖2B分別展示具有一敞開區域/室202之微流體器件200之一封閉體102之一部分之一側視橫截面圖及一俯視橫截面圖，但應瞭解，區域/室202可為具有一更詳細結構(諸如一生長室、一流動區域或一流動通道)之一流體線路元件之部分。此外，微流體器件200可包含其他流體線路元件。例如，微流體器件200可包含複數個生長室及/或一或多個流動區域或流動通道，諸如本文中關於微流體器件100所描述之生長室及/或流動區域或流動通道。可將一DEP構形併入至微流體器件200之任何此等流體線路元件或其選擇部分中。應進一步瞭解，上文所描述或下文將描述之微流體器件組件及系統組件之任何者可併入於微流體器件200中及/或與微流體器件200一起使用。例如，包含上文所描述之控制及監測設備152之系統150可與包含培養基模組160、動力模組162、成像模組164、傾斜模組166及其他模組168之一或多者之微流體器件200一起使用。如圖2A中所見，微流體器件200包含：一支撐結構104，其具有一底部電極204及上覆於底部電極204上之一電極啟動基板206；及一蓋110，其具有一頂部電極210，其中頂部電極210與底部電極204間隔開。頂部電極210及電極啟動基板206界定區域/室202之相對表面。因此，含於區域/室202中之一培養基180提供頂部電極210與電極啟動基板206之間的一電阻連接。圖中亦展示一電源212，其經構形以連接至底部電極204及頂部電極210且在該等電極之間產生產生區域/室202中之DEP力所需之一偏壓電壓。電源212可為(例如)一交流(AC)電源。在某些實施例中，圖2A及圖2B中所繪示之微流體器件200可具有一光學致動之DEP構形。據此，改變來自光源220之光圖案222 (其可由動力模組162控制)可選擇性地啟動及不啟動改變電極啟動基板206之內表面208之區域214處之DEP電極之圖案。(在下文中，具有一DEP構形之一微流體器件之區域214指稱「DEP電極區域」)。如圖2B中所繪示，導引至電極啟動基板206之內表面208上之一光圖案222可照射呈一圖案(諸如一正方形)之選定DEP電極區域214a (展示為白色)。未經照射之DEP電極區域214 (畫有交叉影線)在下文中指稱「暗」DEP電極區域214。通過DEP電極啟動基板206 (即，自底部電極204直至與流動區域106中之培養基180界接之電極啟動基板206之內表面208)之相對電阻抗大於通過各暗DEP電極區域214處之區域/室202中之培養基180 (即，自電極啟動基板206之內表面208至蓋110之頂部電極210)之相對電阻抗。然而，一經照射之DEP電極區域214a展現通過電極啟動基板206之一減小相對阻抗，其小於通過各經照射DEP電極區域214a處之區域/室202中之培養基180之相對阻抗。在啟動電源212之後，前述DEP構形在經照射之DEP電極區域214a與相鄰暗DEP電極區域214之間的流體培養基180中產生一電場梯度，其繼而產生吸引或排斥流體培養基180中之鄰近微小物體(圖中未展示)之局部DEP力。因此，可藉由改變自一光源220投射至微流體器件200中之光圖案222而在區域/室202之內表面208處之諸多不同此等DEP電極區域214處選擇性地啟動及不啟動吸引或排斥流體培養基180中之微小物體之DEP電極。DEP力是否吸引或排斥鄰近微小物體可取決於諸如電源212之頻率及培養基180及/或微小物體(圖中未展示)之介電性質之參數。圖2B中所繪示之經照射DEP電極區域214a之正方形圖案224僅係一實例。可由投影至器件200中之光圖案222照射(且藉此啟動) DEP電極區域214之任何圖案，且可藉由改變或移動光圖案222而重複地改變經照射/經啟動DEP電極區域214之圖案。在一些實施例中，電極啟動基板206可包括一光導材料或由一光導材料組成。在此等實施例中，電極啟動基板206之內表面208可為無特徵的。例如，電極啟動基板206可包括一層氫化非晶矽(a-Si:H)或由一層氫化非晶矽(a-Si:H)組成。a-Si:H可包括(例如)約8%至約40%氫(經計算為100*氫原子之數目/氫原子及矽原子之總數目)。a-Si:H層可具有約500奈米至約2.0微米之一厚度。在此等實施例中，DEP電極區域214可產生於電極啟動基板206之內表面208上之任何位置處且在電極啟動基板206之內表面208上呈根據光圖案222之任何圖案。因此，DEP電極區域214之數目及圖案無需固定，而是可對應於光圖案222。例如，美國專利第RE 44,711號(Wu等人)(最初發表為美國專利第7,612,355號)中已描述具有一DEP構形(其包括諸如上文所討論之一光導層)之微流體器件之實例，該專利之全部內容以引用的方式併入本文中。在其他實施例中，電極啟動基板206可包括一基板，其包括形成半導體積體電路之複數個摻雜層、電絕緣層(或區域)及導電層，諸如半導體領域中所知。例如，電極啟動基板206可包括複數個光電晶體(其包含(例如)橫向雙極光電晶體)，各光電晶體對應於一DEP電極區域214。替代地，電極啟動基板206可包括由光電晶體開關控制之電極(例如導電金屬電極)，其中各此類電極對應於一DEP電極區域214。電極啟動基板206可包含此等光電晶體或光電晶體控制電極之一圖案。例如，該圖案可為配置成列及行之實質上呈正方形之光電晶體或光電晶體控制電極之一陣列，諸如圖2B中所展示。替代地，該圖案可為形成一六方晶格之實質上呈六邊形之光電晶體或光電晶體控制電極之一陣列。無論圖案如何，電路元件可形成電極啟動基板206之內表面208處之DEP電極區域214與底部電極210之間的電連接，且該等電連接(即，光電晶體或電極)可由光圖案222選擇性地啟動及不啟動。當未啟動各電連接時，各電連接可具有高阻抗，使得通過電極啟動基板206 (即，自底部電極204至與區域/室202中之培養基180界接之電極啟動基板206之內表面208)之相對阻抗大於通過對應DEP電極區域214處之培養基180 (即，自電極啟動基板206之內表面208至蓋110之頂部電極210)之相對阻抗。然而，當由光圖案222中之光啟動時，通過電極啟動基板206之相對阻抗小於通過各經照射DEP電極區域214處之培養基180之相對阻抗，藉此啟動對應DEP電極區域214處之DEP電極，如上文所討論。因此，可依由光圖案222判定之一方式在區域/室202中之電極啟動基板206之內表面208處之諸多不同DEP電極區域214處選擇性地啟動及不啟動吸引或排斥培養基180中之微小物體(圖中未展示)之DEP電極。例如，美國專利第7,956,339號(Ohta等人)中已描述具有包括光電晶體之電極啟動基板之微流體器件之實例(例如，參閱圖21及圖22中所繪示之器件300及其描述)，該專利之全部內容以引用的方式併入本文中。例如，美國專利申請案第2014/0124370號(Short等人)中已描述具有包括由光電晶體開關控制之電極之電極啟動基板之微流體器件之實例(例如，參閱全部圖式中所繪示之器件200、400、500、600及900及其描述)，該案之全部內容以引用的方式併入本文中。在一DEP構形之微流體器件之一些實施例中，頂部電極210係封閉體102之一第一壁(或蓋110)之部分，且電極啟動基板206及底部電極204係封閉體102之一第二壁(或支撐結構104)之部分。區域/室202可介於第一壁與第二壁之間。在其他實施例中，電極210係第二壁(或支撐結構104)之部分且電極啟動基板206及/或電極210之一或兩者係第一壁(或蓋110)之部分。而且，光源220可替代地用以自下方照射封閉體102。就具有一DEP構形之圖2A至圖2B之微流體器件200而言，動力模組162可藉由將一光圖案222投影至器件200中以啟動一圖案(例如正方形圖案224)(其包圍且捕獲之區域/室202中之培養基180中之一微小物體(圖中未展示))中之電極啟動基板206之內表面208之DEP電極區域214a處之一第一組之一或多個DEP電極而選擇該微小物體。接著，動力模組162可藉由相對於器件200移動光圖案222以啟動DEP電極區域214處之一第二組之一或多個DEP電極而移動該捕獲之微小物體。替代地，可相對於光圖案222移動器件200。在其他實施例中，微流體器件200可具有一DEP構形，其不依賴於電極啟動基板206之內表面208處之DEP電極之光啟動。例如，電極啟動基板206可包括可選擇性地定址及供能之電極，其經定位成相對於包含至少一電極(例如蓋110)之一表面。開關(例如一半導體基板中之電晶體開關)可選擇性地接通及切斷以啟動或不啟動DEP電極區域214處之DEP電極，藉此在啟動DEP電極附近產生作用於區域/室202中之一微小物體(圖中未展示)之一凈DEP力。取決於諸如電源212之頻率及區域/室202中之培養基(圖中未展示)及/或微小物體之介電性質之特性，DEP力可吸引或排斥一鄰近微小物體。可藉由選擇性地啟動及不啟動一組DEP電極(例如，在形成一正方形圖案224之一組DEP電極區域214處)而捕集區域/室202中之一或多個微小物體且使其在區域/室202內移動。圖1中之動力模組162可控制此等開關且因此啟動及不啟動DEP電極之個別者以選擇、捕集及移動區域/室202周圍之特定微小物體(圖中未展示)。此項技術中已知且(例如)美國專利第6,294,063號(Becker等人)及第6,942,776號(Medoro)中已描述具有包含可選擇性定址及供能之電極之一DEP構形之微流體器件，該等專利之全部內容以引用的方式併入本文中。作為又一實例，微流體器件200可具有一電潤濕(EW)構形，其可替換DEP構形或可定位於與具有DEP構形之部分分離之微流體器件200之一部分中。EW構形可為一光電潤濕構形或一介電潤濕潤(EWOD)構形，該兩個構形在此項技術中係已知的。在一些EW構形中，支撐結構104具有夾於一介電層(圖中未展示)與底部電極204之間的一電極啟動基板206。該介電層可包括一疏水材料及/或可塗佈有一疏水材料。針對具有一EW構形之微流體器件200，支撐結構104之內表面208係該介電層或其疏水塗層之內表面。介電層(圖中未展示)可包括一或多個氧化層，且可具有約50奈米至約250奈米(例如，約125奈米至約175奈米)之一厚度。在某些實施例中，介電層可包括一層氧化物，諸如一金屬氧化物(例如氧化鋁或氧化鉿)。在某些實施例中，介電層可包括除一金屬氧化物之外之一介電材料，諸如氧化矽或氮化物。無論準確組合物及厚度如何，介電層可具有約10千歐姆至約50千歐姆之一阻抗。在一些實施例中，向內面向區域/室202之介電層之表面塗佈有一疏水材料。該疏水材料可包括(例如)氟化碳分子。氟化碳分子之實例包含全氟聚合物，諸如聚四氟乙烯(例如TEFLON®)或聚(2,3-二氟亞甲基-全氟四氫呋喃)(例如CYTOP™)。組成該疏水材料之分子可共價鍵聯至介電層之表面。例如，該疏水材料之分子可藉由一鍵聯子(諸如矽氧烷基、膦酸基或硫醇基)而共價鍵聯至介電層之表面。因此，在一些實施例中，該疏水材料可包括烷基封端之矽氧烷、烷基封端之膦酸或烷基封端之硫醇。烷基可為長鏈碳氫化合物(例如，具有至少10個碳或至少16個、18個、20個、22個或22個以上碳之一鏈)。替代地，氟化(或全氟化)碳鏈可用於替換烷基。因此，例如，該疏水材料可包括氟烷基封端之矽氧烷、氟烷基封端之膦酸或氟烷基封端之硫醇。在一些實施例中，該疏水塗層具有約10奈米至約50奈米之一厚度。在其他實施例中，該疏水塗層具有小於10奈米(例如，小於5奈米或約1.5奈米至約3.0奈米)之一厚度。在一些實施例中，具有一電潤濕構形之一微流體器件200之蓋110亦塗佈有一疏水材料(圖中未展示)。該疏水材料可為用以塗佈支撐結構104之介電層之相同疏水材料，且該疏水塗層可具有實質上相同於支撐結構104之介電層上之疏水塗層之厚度的一厚度。而且，蓋110可包括依支撐結構104之方式夾於一介電層與頂部電極210之間的一電極啟動基板206。電極啟動基板206及蓋110之介電層可具有相同於支撐結構104之電極啟動基板206及介電層之組合物及/或尺寸。因此，微流體器件200可具有兩個電潤濕表面。在一些實施例中，電極啟動基板206可包括一光導材料，諸如上文所描述。據此，在某些實施例中，電極啟動基板206可包括一層氫化非晶矽(a-Si:H)或由一層氫化非晶矽(a-Si:H)組成。例如，a-Si:H可包括約8%至40%氫(經計算為100*氫原子之數目/氫原子及矽原子之總數目)。a-Si:H層可具有約500奈米至約2.0微米之一厚度。替代地，電極啟動基板206可包括由光電晶體開關控制之電極(例如導電金屬電極)，如上文所描述。此項技術中已知及/或可使用此項技術中已知之電極啟動基板來建構具有一光電潤濕構形之微流體器件。例如，美國專利第6,958,132號(Chiou等人)(該專利之全部內容以引用的方式併入本文中)揭示具有一光導材料(諸如a-Si:H)之光電潤濕構形，而上文所引用之美國專利公開案第2014/0124370號(Short等人)揭示具有由光電晶體開關控制之電極之電極啟動基板。因此，微流體器件200可具有一光電潤濕構形，且光圖案222可用以啟動電極啟動基板206中之光導EW區域或光響應EW電極。電極啟動基板206之此等啟動EW區域或EW電極可在支撐結構104之內表面208 (即，上覆介電層或其疏水塗層之內表面)處產生一電潤濕力。可藉由改變入射於電極啟動基板206上之光圖案222 (或相對於光源220移動微流體器件200)而使接觸支撐結構104之內表面208之液滴(例如，其含有一水培養基、溶液或溶劑)移動通過存在於區域/室202中之一不溶混流體(例如一油培養基)。在其他實施例中，微流體器件200可具有一EWOD構形，且電極啟動基板206可包括不依賴於用於啟動之光之可選擇性定址及供能之電極。因此，電極啟動基板206可包含此等電潤濕(EW)電極之一圖案。例如，該圖案可為配置成列及行之實質上呈正方形之EW電極之一陣列，諸如圖2B中所展示。替代地，該圖案可為形成一六方晶格之實質上呈六邊形之EW電極之一陣列。無論圖案如何，可由電開關(例如一半導體基板中之電晶體開關)選擇性地啟動(或不啟動)EW電極。可藉由選擇性地啟動及不啟動電極啟動基板206中之EW電極而使接觸上覆介電層或其疏水塗層之內表面208之液滴(圖中未展示)在區域/室202內移動。圖1中之動力模組162可控制此等開關且因此啟動及不啟動個別EW電極來選擇及移動區域/室202周圍之特定液滴。此項技術中已知且(例如)美國專利第8,685,344號(Sundarsan等人)中已描述具有一EWOD構形(其具有可選擇性定址及供能之電極)之微流體器件，該專利之全部內容以引用的方式併入本文中。無論微流體器件200之構形如何，一電源212可用以提供對微流體器件200之電路供電之一電位(例如一AC電壓電位)。電源212可相同於圖1中所參考之電源192或可為圖1中所參考之電源192之一組件。電源212可經構形以將一AC電壓及/或電流提供至頂部電極210及底部電極204。針對一AC電壓，電源212可提供一頻率範圍及一平均或峰值電力(例如電壓或電流)範圍，其足以產生足夠強之凈DEP力(或電潤濕力)來捕集及移動區域/室202中之微小物體(圖中未展示)(如上文所討論)及/或改變區域/室202中之支撐結構104之內表面208 (即，介電層及/或介電層上之疏水塗層)之濕潤性質(亦如上文所討論)。此等頻率範圍及平均或峰值電力範圍在此項技術中係已知的。例如，參閱美國專利第6,958,132號(Chiou等人)、美國專利第RE44,711號(Wu等人)(最初發表為美國專利第7,612,355號)及美國專利申請公開案第US2014/0124370號(Short等人)、第US2015/0306598號(Khandros等人)及第US2015/0306599號(Khandros等人)。 生長室圖2C及圖2D中所描繪之微流體器件240內展示通用生長室244、246及248之非限制性實例。各生長室244、246及248可包括一隔離結構250，其界定一隔離區域258及將隔離區域258流體地連接至一通道122之一連接區域254。連接區域254可包括至通道122之一近端開口252及至隔離區域258之一遠端開口256。連接區域254可經構形使得自通道122流動至生長室244、246、248中之一流體培養基(圖中未展示)之一流動之最大滲透深度不延伸至隔離區域258中。因此，歸因於連接區域254，安置於一生長室244、246、248之一隔離區域258中之一微小物體(圖中未展示)或其他材料(圖中未展示)可因此與通道122中之培養基180之一流動隔離且實質上不受通道122中之培養基180之一流動影響。因此，通道122可為一掃及區域之一實例，且生長室244、246、248之隔離區域258可為未掃及區域之實例。如本文所提及，通道122及生長室244、246、248可經構形以含有一或多個流體培養基180。在圖2C至圖2D所展示之實例中，端口242連接至通道122且允許將一流體培養基180引入至微流體器件240中或自微流體器件240移除一流體培養基180。在引入流體培養基180之前，微流體器件可預充有諸如二氧化碳氣體之一氣體。一旦微流體器件240含有流體培養基180，則可選擇性地產生且停止通道122中之流體培養基180之流260。例如，如圖中所展示，可將端口242安置於通道122之不同位置(例如相對端)處，且可自用作一入口之一端口242至用作一出口之另一端口242產生培養基之一流260。圖2E繪示根據本發明之一生長室244之一實例之一詳細圖。圖中亦展示微小物體270之實例。眾所周知，一微流體通道122中之流體培養基180之一流260通過生長室244之一近端開口252可致使培養基180之一副流262進入及/或流出生長室244。為使一生長室244之隔離區域258中之微小物體270與副流262隔離，生長室244之連接區域254之長度L _con(即，自近端開口252至遠端開口256)應大於副流262進入連接區域254之滲透深度D _p。副流262之滲透深度D _p取決於在通道122中流動之流體培養基180之速度及與通道122之構形及至通道122之連接區域254之近端開口252相關之各種參數。針對一給定微流體器件，通道122及開口252之構形將為固定的，而通道122中之流體培養基180之流260之速率將為可變的。據此，針對各生長室244，可識別通道122中之流體培養基180之流260之一最大速度Vmax，其確保副流262之滲透深度D _p不超過連接區域254之長度L _con。只要通道122中之流體培養基180之流260之速率不超過最大速度Vmax，則所得副流262可受限於通道122及連接區域254且保持不進入隔離區域258。因此，通道122中之培養基180之流260不會自隔離區域258帶出微小物體270。確切而言，無論通道122中之流體培養基180之流260如何，定位於隔離區域258中之微小物體270將留在隔離區域258中。而且，只要通道122中之培養基180之流260之速率不超過Vmax，則通道122中之流體培養基180之流260不會將混雜粒子(例如微粒及/或奈米粒子)自通道122移動至一生長室244之隔離區域258中。因此，使連接區域254之長度L _con大於副流262之最大滲透深度D _p可防止來自通道122或另一生長室(例如圖2D中之生長室246、248)之混雜粒子污染一生長室244。因為通道122及生長室244、246、248之連接區域254會受通道122中之培養基180之流260影響，所以通道122及連接區域254可被視作微流體器件240之掃及(或流動)區域。在另一方面，生長室244、246、248之隔離區域258可被視作未掃及(或非流動)區域。例如，通道122中之一第一流體培養基180中之組分(圖中未展示)可實質上僅藉由第一培養基180之組分自通道122擴散通過連接區域254且進入隔離區域258中之一第二流體培養基280而與隔離區域258中之第二流體培養基280混合。類似地，隔離區域258中之第二培養基280之組分(圖中未展示)可實質上僅藉由第二培養基280之組分自隔離區域258擴散通過連接區域254且進入通道122中之第一培養基180而與通道122中之第一培養基180混合。第一培養基180可為相同於或不同於第二培養基280之一培養基。而且，第一培養基180及第二培養基280可起初相同，接著變為不同(例如，透過由隔離區域258中之一或多個細胞調節第二培養基280，或藉由改變流動通過通道122之培養基180)。由通道122中之流體培養基180之流260所致之副流262之最大滲透深度D _p可取決於如上文所提及之若干參數。此等參數之實例包含通道122之形狀(例如，通道可將培養基導引至連接區域254中，自連接區域254轉走培養基，或在實質上垂直於至通道122之連接區域254之近端開口252之一方向上導引培養基)、近端開口252處之通道122之一寬度W _ch(或橫截面積)、近端開口252處之連接區域254之一寬度W _con(或橫截面積)、通道122中之流體培養基180之流260之速度V、第一培養基180及/或第二培養基280之黏度、或其類似者。在一些實施例中，通道122及生長室244、246、248之尺寸可相對於通道122中之流體培養基180之流260之向量而定向如下：通道寬度W _ch(或通道122之橫截面積)可實質上垂直於培養基180之流260；開口252處之連接區域254之寬度W _con(或橫截面積)可實質上平行於通道122中之培養基180之流260；及/或連接區域之長度L _con可實質上垂直於通道122中之培養基180之流260。上述定向僅為實例，且通道122及生長室244、246、248之相對位置可在相對於彼此之其他定向上。如圖2E中所繪示，自近端開口252至遠端開口256之連接區域254之寬度W _con可為均勻的。因此，遠端開口256處之連接區域254之寬度W _con可在本文中針對近端開口252處之連接區域254之寬度W _con所識別之範圍之任何者內。替代地，遠端開口256處之連接區域254之寬度W _con可大於近端開口252處之連接區域254之寬度W _con。如圖2E中所繪示，遠端開口256處之隔離區域258之寬度可實質上相同於近端開口252處之連接區域254之寬度W _con。因此，遠端開口256處之隔離區域258之寬度可在本文中針對近端開口252處之連接區域254之寬度W _con所識別之範圍之任何者內。替代地，遠端開口256處之隔離區域258之寬度可大於或小於近端開口252處之連接區域254之寬度W _con。而且，遠端開口256可小於近端開口252且連接區域254之寬度W _con可在近端開口252與遠端開口256之間變窄。例如，可使用各種不同幾何形狀來使連接區域254在近端開口與遠端開口之間變窄(例如，斜裁連接區域，斜切連接區域)。進一步言之，連接區域254之任何部分或子部分可變窄(例如相鄰於近端開口252之連接區域之一部分)。圖4A至圖4C描繪含有一微流體線路432及流動通道434之一微流體器件400之另一例示性實施例，微流體器件400、微流體線路432及流動通道434係圖1之各自微流體器件100、線路132及通道134之變體。微流體器件400亦具有複數個生長室436，其係上文所描述之生長室124、126、128、130、244、246或248之額外變體。特定言之，應瞭解，圖4A至圖4C中所展示之器件400之生長室436可替換器件100、200、240及290中之上文所描述之生長室124、126、128、130、244、246或248之任何者。同樣地，微流體器件400係微流體器件100之另一變體且亦可具有相同於或不同於上文所描述之微流體器件100、200、240、290之DEP構形以及本文中所描述之其他微流體系統組件之任何者。圖4A至圖4C之微流體器件400包括一支撐結構(圖4A至圖4C中不可見，但可相同於或大體上類似於圖1中所描繪之器件100之支撐結構104)、一微流體線路結構412及一蓋(圖4A至圖4C中不可見，但可相同於或大體上類似於圖1中所描繪之器件100之蓋110)。微流體線路結構412包含一框架414及微流體線路材料416，其等可相同於或大體上類似於圖1中所展示之器件100之框架114及微流體線路材料116。如圖4A中所展示，由微流體線路材料416界定之微流體線路432可包括多個生長室436流體地連接至其之多個通道434 (圖中展示兩個，但可存在更多個)。各生長室436可包括一隔離結構446、隔離結構446內之一隔離區域444、及一連接區域442。連接區域442自通道434處之一近端開口472至隔離結構436處之一遠端開口474將通道434流體地連接至隔離區域444。一般而言，根據圖2D及圖2E之上述討論，一通道434中之一第一流體培養基402之一流482可產生自通道434進入生長室436之各自連接區域442及/或離開生長室436之各自連接區域442之第一培養基402之副流484。如圖4B中所繪示，各生長室436之連接區域442大體上包含延伸於至一通道434之近端開口472與至一隔離結構446之遠端開口474之間的區域。連接區域442之長度L _con可大於副流484之最大滲透深度D _p，在該情況中，副流484將延伸至連接區域442中且未被轉向隔離區域444 (如圖4A中所展示)。替代地，如圖4C中所繪示，連接區域442可具有小於最大滲透深度D _p之一長度L _con，在該情況中，副流484將延伸通過連接區域442且被轉向隔離區域444。在此後一情況中，連接區域442之長度L _c1及L _c2之總和大於最大滲透深度D _p，使得副流484不會延伸至隔離區域444中。無論連接區域442之長度L _con大於滲透深度D _p或連接區域442之長度L _c1及L _c2之總和大於滲透深度D _p，通道434中之一第一培養基402之一流482 (其不超過一最大速度V _max)將產生具有一滲透深度D _p之一副流，且一生長室436之隔離區域444中之微小物體(圖中未展示，但可相同於或大體上類似於圖2E中所展示之微小物體270)不會被通道434中之第一培養基402之一流482帶出隔離區域444。通道434中之流482亦不會將混雜材料(圖中未展示)自通道434帶入至一生長室436之隔離區域444中。因而，擴散係通道434中之一第一培養基402中之組分可自通道434移動至一生長室436之一隔離區域444中之一第二培養基404中之唯一機制。同樣地，擴散係一生長室436之一隔離區域444中之一第二培養基404中之組分可自隔離區域444移動至通道434中之一第一培養基402之唯一機制。第一培養基402可為相同於第二培養基404之培養基，或第一培養基402可為不同於第二培養基404之一培養基。替代地，第一培養基402及第二培養基404可起初相同，接著變為不同，例如，透過由隔離區域444中之一或多個細胞調節第二培養基，或藉由改變流動通過通道434之培養基。如圖4B中所繪示，通道434中之通道434之寬度W _ch(即，橫向於流動通過通道之一流體培養基之方向(由圖4A中之箭頭482指示)而取得)可實質上垂直於近端開口472之一寬度W _con1且因此實質上平行於遠端開口474之一寬度W _con2。然而，近端開口472之寬度W _con1及遠端開口474之寬度W _con2無需實質上彼此垂直。例如，近端開口472之寬度W _con1定向於其上之一軸(圖中未展示)與遠端開口474之寬度W _con2定向於其上之另一軸之間的一角度可不成直角且因此不為90°。替代定向角之實例包含下列範圍之任何者內之角度：自約30°至約90°、自約45°至約90°、自約60°至約90°或其類似者。在生長室(例如124、126、128、130、244、246、248或436)之各種實施例中，隔離區域(例如258或444)經構形以含有複數個微小物體。在其他實施例中，隔離區域可經構形以僅含有1個、2個、3個、4個、5個或類似相對較小數目個微小物體。據此，一隔離區域之體積可為(例如)至少3×10 ³立方微米、6×10 ³立方微米、9×10 ³立方微米、1×10 ⁴立方微米、2×10 ⁴立方微米、4×10 ⁴立方微米、8×10 ⁴立方微米、1×10 ⁵立方微米、2×10 ⁵立方微米、4×10 ⁵立方微米、8×10 ⁵立方微米、1×10 ⁶立方微米、2×10 ⁶立方微米、4×10 ⁶立方微米、6×10 ⁶立方微米、1×10 ⁷立方微米、2×10 ⁷立方微米、4×10 ⁷立方微米、6×10 ⁷立方微米、1×10 ⁸立方微米或更大。在生長室之各種實施例中，一近端開口(例如252、472)處之通道122、434之寬度W _ch可在下列範圍之任何者內：50微米至1000微米、50微米至500微米、50微米至400微米、50微米至300微米、50微米至250微米、50微米至200微米、50微米至150微米、50微米至100微米、70微米至500微米、70微米至400微米、70微米至300微米、70微米至250微米、70微米至200微米、70微米至150微米、90微米至400微米、90微米至300微米、90微米至250微米、90微米至200微米、90微米至150微米、100微米至300微米、100微米至250微米、100微米至200微米、100微米至150微米及100微米至120微米。上述範圍僅為實例，且通道122、434之寬度W _ch可在其他範圍(例如由上文所列之端點之任何者界定之一範圍)內。而且，通道122、434之W _ch可經選定為在除一生長室之一近端開口處之外之通道之區域中之此等區域之任何者內。在一些實施例中，一生長室具有約30微米至約200微米或約50微米至約150微米之一橫截面高度。在一些實施例中，生長室具有約100,000平方微米至約2,500,000平方微米或約200,000平方微米至約2,000,000平方微米之一橫截面積。在一些實施例中，一連接區域具有匹配對應生長室之橫截面高度之一橫截面高度。在一些實施例中，連接區域具有約50微米至約500微米或約100微米至約300微米之一橫截面寬度。在生長室之各種實施例中，一近端開口252、472處之通道122、434之高度H _ch可在下列範圍之任何者內：20微米至100微米、20微米至90微米、20微米至80微米、20微米至70微米、20微米至60微米、20微米至50微米、30微米至100微米、30微米至90微米、30微米至80微米、30微米至70微米、30微米至60微米、30微米至50微米、40微米至100微米、40微米至90微米、40微米至80微米、40微米至70微米、40微米至60微米或40微米至50微米。上述範圍僅為實例，且通道122、434之高度H _ch可在其他範圍(例如由上文所列之端點之任何者界定之一範圍)內。通道122、434之高度H _ch可經選擇為在除一生長室之一近端開口處之外之通道之區域中之此等範圍之任何者內。在生長室之各種實施例中，一近端開口252、472處之通道122、434之一橫截面積可在下列範圍之任何者內：500平方微米至50,000平方微米、500平方微米至40,000平方微米、500平方微米至30,000平方微米、500平方微米至25,000平方微米、500平方微米至20,000平方微米、500平方微米至15,000平方微米、500平方微米至10,000平方微米、500平方微米至7,500平方微米、500平方微米至5,000平方微米、1,000平方微米至25,000平方微米、1,000平方微米至20,000平方微米、1,000平方微米至15,000平方微米、1,000平方微米至10,000平方微米、1,000平方微米至7,500平方微米、1,000平方微米至5,000平方微米、2,000平方微米至20,000平方微米、2,000平方微米至15,000平方微米、2,000平方微米至10,000平方微米、2,000平方微米至7,500平方微米、2,000平方微米至6,000平方微米、3,000平方微米至20,000平方微米、3,000平方微米至15,000平方微米、3,000平方微米至10,000平方微米、3,000平方微米至7,500平方微米或3,000平方微米至6,000平方微米。上述範圍僅為實例，且一近端開口252、472處之通道122之橫截面積可在其他範圍(例如由上文所列之端點之任何者界定之一範圍)內。在生長室之各種實施例中，連接區域254、442之長度L _con可在下列範圍之任何者內：1微米至200微米、5微米至150微米、10微米至100微米、15微米至80微米、20微米至60微米、20微米至500微米、40微米至400微米、60微米至300微米、80微米至200微米及100微米至150微米。上述範圍僅為實例，且一連接區域254、442之長度L _con可在不同於上述實例之一範圍(例如由上文所列之端點之任何者界定之一範圍)內。在生長室之各種實施例中，一近端開口252處之一連接區域254、442之寬度W _con可在下列範圍之任何者內：20微米至500微米、20微米至400微米、20微米至300微米、20微米至200微米、20微米至150微米、20微米至100微米、20微米至80微米、20微米至60微米、30微米至400微米、30微米至300微米、30微米至200微米、30微米至150微米、30微米至100微米、30微米至80微米、30微米至60微米、40微米至300微米、40微米至200微米、40微米至150微米、40微米至100微米、40微米至80微米、40微米至60微米、50微米至250微米、50微米至200微米、50微米至150微米、50微米至100微米、50微米至80微米、60微米至200微米、60微米至150微米、60微米至100微米、60微米至80微米、70微米至150微米、70微米至100微米及80微米至100微米。上述範圍僅為實例，且一近端開口252處之一連接區域254、442之寬度W _con可不同於上述實例(例如由上文所列之端點之任何者界定之一範圍)。在生長室之各種實施例中，一近端開口252、472處之一連接區域254、442之寬度W _con可在下列範圍之任何者內：2微米至35微米、2微米至25微米、2微米至20微米、2微米至15微米、2微米至10微米、2微米至7微米、2微米至5微米、2微米至3微米、3微米至25微米、3微米至20微米、3微米至15微米、3微米至10微米、3微米至7微米、3微米至5微米、3微米至4微米、4微米至20微米、4微米至15微米、4微米至10微米、4微米至7微米、4微米至5微米、5微米至15微米、5微米至10微米、5微米至7微米、6微米至15微米、6微米至10微米、6微米至7微米、7微米至15微米、7微米至10微米、8微米至15微米及8微米至10微米。上述範圍僅為實例，且一近端開口252、472處之一連接區域254、442之寬度W _con可不同於上述實例(例如由上文所列之端點之任何者界定之一範圍)。在生長室之各種實施例中，近端開口252、472處之一連接區域254、442之長度L _con與連接區域254、442之一寬度W _con之一比率可大於或等於下列比率之任何者：0.5、1.0、1.5、2.0、2.5、3.0、3.5、4.0、4.5、5.0、6.0、7.0、8.0、9.0、10.0或更大。上述比率僅為實例，且近端開口252、472處之一連接區域254之長度L _con與連接區域254、442之一寬度W _con之比率可不同於上述實例。在微流體器件100、200、240、290、400之各種實施例中，V _max可經設定為約0.2微升/秒、約0.3微升/秒、約0.4微升/秒、約0.5微升/秒、約0.6微升/秒、約0.7微升/秒、約0.8微升/秒、約0.9微升/秒、約1.0微升/秒、約1.1微升/秒、約1.2微升/秒、約1.3微升/秒、約1.4微升/秒或約1.5微升/秒。在一些其他實施例中，替代地，V _max可經設定為約0.2微升/秒、約0.3微升/秒、約0.4微升/秒、約0.5微升/秒、約0.6微升/秒、約0.7微升/秒、約0.8微升/秒、約0.9微升/秒、約1.0微升/秒、約1.1微升/秒、約1.2微升/秒、約1.3微升/秒、約1.4微升/秒、約1.5微升/秒、約1.6微升/秒、約1.7微升/秒、約1.8微升/秒、約1.9微升/秒、約2.0微升/秒、約2.1微升/秒、約2.2微升/秒、約2.3微升/秒、約2.4微升/秒或約2.5微升/秒。在其他實施例中，V _max可經設定為或大致設定為2.0微升/秒、2.2微升/秒、2.4微升/秒、2.6微升/秒、2.8微升/秒、3.0微升/秒、3.2微升/秒、3.4微升/秒、3.6微升/秒、3.8微升/秒、4.0微升/秒、4.2微升/秒、4.4微升/秒、4.6微升/秒、4.8微升/秒、5.0微升/秒、6.0微升/秒、7.0微升/秒、8.0微升/秒或9.0微升/秒。在具有生長室之微流體器件之各種實施例中，一生長室之一隔離區域258、444之體積可為(例如)至少3×10 ³立方微米、6×10 ³立方微米、9×10 ³立方微米、1×10 ⁴立方微米、2×10 ⁴立方微米、4×10 ⁴立方微米、8×10 ⁴立方微米、1×10 ⁵立方微米、2×10 ⁵立方微米、4×10 ⁵立方微米、8×10 ⁵立方微米、1×10 ⁶立方微米、2×10 ⁶立方微米、4×10 ⁶立方微米、6×10 ⁶立方微米或更大。在具有生長室之微流體器件之各種實施例中，一生長室之體積可為約5×10 ³立方微米、約7×10 ³立方微米、約1×10 ⁴立方微米、約3×10 ⁴立方微米、約5×10 ⁴立方微米、約8×10 ⁴立方微米、約1×10 ⁵立方微米、約2×10 ⁵立方微米、約4×10 ⁵立方微米、約6×10 ⁵立方微米、約8×10 ⁵立方微米、約1×10 ⁶立方微米、約2×10 ⁶立方微米、約4×10 ⁶立方微米、約8×10 ⁶立方微米、約1×10 ⁷立方微米、約3×10 ⁷立方微米、約5×10 ⁷立方微米或約8×10 ⁷立方微米或更大。在一些實施例中，微流體器件具有其中可維持不超過1×10 ²個生物細胞之生長室，且一生長室之體積可不超過2×10 ⁶立方微米。在一些實施例中，微流體器件具有其中可維持不超過1×10 ²個生物細胞之生長室，且一生長室可不超過4×10 ⁵立方微米。在其他實施例中，微流體器件具有其中可維持不超過50個生物細胞之生長室，一生長室可不超過4×10 ⁵立方微米。在各種實施例中，微流體器件具有如同本文中所討論之實施例之任何者般構形之生長室，其中微流體器件具有約100個至約500個生長室、約200個至約1000個生長室、約500個至約1500個生長室、約1000個至約2000個生長室或約1000個至約3500個生長室。在一些其他實施例中，微流體器件具有如同本文中所討論之實施例之任何者般構形之生長室，其中微流體器件具有約1500個至約3000個生長室、約2000個至約3500個生長室、約2500個至約4000個生長室、約3000個至約4500個生長室、約3500個至約5000個生長室、約4000個至約5500個生長室、約4500個至約6000個生長室、約5000個至約6500個生長室、約5500個至約7000個生長室、約6000個至約7500個生長室、約6500個至約8000個生長室、約7000個至約8500個生長室、約7500個至約9000個生長室、約8000個至約9500個生長室、約8500個至約10,000個生長室、約9000個至約10,500個生長室、約9500個至約11,000個生長室、約10,000個至約11,500個生長室、約10,500個至約12,000個生長室、約11,000個至約12,500個生長室、約11,500個至約13,000個生長室、約12,000個至約13,500個生長室、約12,500個至約14,000個生長室、約13,000個至約14,500個生長室、約13,500個至約15,000個生長室、約14,000個至約15,500個生長室、約14,500個至約16,000個生長室、約15,000個至約16,500個生長室、約15,500個至約17,000個生長室、約16,000個至約17,500個生長室、約16,500個至約18,000個生長室、約17,000個至約18,500個生長室、約17,500個至約19,000個生長室、約18,000個至約19,500個生長室、約18,500個至約20,000個生長室、約19,000個至約20,500個生長室、約19,500個至約21,000個生長室或約20,000個至約21,500個生長室。圖2F繪示根據一實施例之一微流體器件290。圖2F中所繪示之微流體器件290係一微流體器件100之一典型圖。實際上，微流體器件290及其組成線路元件(例如通道122及生長室128)將具有本文中所討論之尺寸。圖2F中所繪示之微流體線路120具有兩個端口107、四個不同通道122及四個不同流動路徑106。微流體器件290進一步包括朝向各通道122敞開之複數個生長室。在圖2F所繪示之微流體器件中，生長室具有類似於圖2E中所繪示之圍欄之一幾何形狀，且因此具有連接區域及隔離區域兩者。據此，微流體線路120包含掃及區域(例如通道122及副流262之最大滲透深度D _p內之連接區域254之部分)及未掃及區域(例如隔離區域258及不在副流262之最大滲透深度D _p內之連接區域254之部分)兩者。圖3A及圖3B展示根據本發明之可用以操作及觀測微流體器件(例如100、200、240、290)之系統150之各種實施例。如圖3A中所繪示，系統150可包含經構形以保持一微流體器件100 (圖中未展示)或本文中所描述之任何其他微流體器件之一結構(「巢套」) 300。巢套300可包含能夠與微流體器件360 (例如一光學致動之電動器件100)界接且提供自電源192至微流體器件360之電連接之一插座302。巢套300可進一步包含一整合電信號產生子系統304。電信號產生子系統304可經構形以將一偏壓電壓供應至插座302，使得當由插座302保持微流體器件360時，橫跨微流體器件360中之一對電極而施加該偏壓電壓。因此，電信號產生子系統304可為電源192之部分。將一偏壓電壓施加於微流體器件360之能力不意謂將在由插座302保持微流體器件360時始終施加一偏壓電壓。確切而言，在大多數情況中，將間歇地施加偏壓電壓，例如，僅在需要促進在微流體器件360中產生電動力(諸如介電泳或電潤濕)時施加偏壓電壓。如圖3A中所繪示，巢套300可包含一印刷電路板總成(PCBA) 320。電信號產生子系統304可安裝於PCBA 320上且電整合至PCBA 320中。例示性巢套300亦包含安裝於PCBA 320上之插座302。通常，電信號產生子系統304將包含一波形產生器(圖中未展示)。電信號產生子系統304可進一步包含經構形以放大自該波形產生器接收之一波形之一示波器(圖中未展示)及/或一波形放大電路(圖中未展示)。該示波器(若存在)可經構形以量測供應至由插座302保持之微流體器件360之波形。在某些實施例中，該示波器量測接近於微流體器件360 (且遠離於該波形產生器)之一位置處之波形以因此確保量測實際上施加於器件之波形時之較大精確度。例如，可將自該示波器量測獲得之資料作為回饋提供至該波形產生器，且該波形產生器可經構形以基於此回饋而調整其輸出。一適當組合之波形產生器及示波器之一實例係Red Pitaya™。在某些實施例中，巢套300進一步包括一控制器308，諸如用以感測及/或控制電信號產生子系統304之一微處理器。適合微處理器之實例包含Arduino™微處理器，諸如Arduino Nano™。控制器308可用以執行功能及分析或可與一外部主控制器154 (如圖1中所展示)通信以執行功能及分析。在圖3A所繪示之實施例中，控制器308透過一介面310 (例如一插頭或連接器)而與一主控制器154通信。在一些實施例中，巢套300可包括一電信號產生子系統304，其包括一Red Pitaya™波形產生器/示波器單元(「Red Pitaya單元」)及一波形放大電路，該波形放大電路放大由該Red Pitaya單元產生之波形且將放大電壓傳至微流體器件100。在一些實施例中，該Red Pitaya單元經構形以量測微流體器件360處之放大電壓且接著根據需要而調整其自身輸出電壓，使得微流體器件360處之量測電壓係所要值。在一些實施例中，該波形放大線路可具有由安裝於PCBA 320上之一對DC-DC轉換器產生之一+6.5伏特至-6.5伏特電源供應器以導致微流體器件100處之高達13 Vpp之一信號。如圖3A中所繪示，巢套300可進一步包含一熱控制子系統306。熱控制子系統306可經構形以調節由巢套300保持之微流體器件360之溫度。例如，熱控制子系統306可包含一帕耳帖(Peltier)熱電器件(圖中未展示)及一冷卻單元(圖中未展示)。該帕耳帖熱電器件可具有經構形以與微流體器件360之至少一表面界接之一第一表面。該冷卻單元可為(例如)一冷卻塊(圖中未展示)，諸如一液冷鋁塊。該帕耳帖熱電器件之一第二表面(例如相對於該第一表面之一表面)可經構形以與此一冷卻塊之一表面界接。該冷卻塊可連接至經構形以使冷卻流體循環通過該冷卻塊之一流體路徑330。在圖3A所繪示之實施例中，巢套300包括一入口332及一出口334來自一外部儲液器(圖中未展示)接收冷卻流體，將該冷卻流體引入至流體路徑330中且通過該冷卻塊，且接著使該冷卻流體返回至該外部儲液器。在一些實施例中，該帕耳帖熱電器件、該冷卻單元及/或流體路徑330可安裝於巢套300之一罩殼340上。在一些實施例中，熱控制子系統306經構形以調節該帕耳帖熱電器件之溫度以便達到微流體器件360之一目標溫度。例如，可藉由一熱電電源供應器(諸如一Pololu™熱電電源供應器(Pololu Robotics and Electronics公司))而達成該帕耳帖熱電器件之溫度調節。熱控制子系統306可包含一回饋電路，諸如由一類比電路提供之一溫度值。替代地，該回饋電路可由一數位電路提供。在一些實施例中，巢套300可包含具有一回饋電路(其係一類比分壓器電路(圖中未展示))之一熱控制子系統306，其包含一電阻器(例如，具有1千歐姆+/-0.1%之電阻、+/-0.02 ppm/C0之溫度係數)及一NTC熱阻器(例如，具有1千歐姆+/-0.01%之標稱電阻)。在一些例項中，熱控制子系統306自回饋電路量測電壓且接著將計算溫度值用作為至一板上PID控制迴路演算法之輸入。來自該PID控制迴路演算法之輸出可驅動(例如)一Pololu™馬達驅動器(圖中未展示)上之一方向接針及一脈寬調變信號接針兩者來致動熱電電源供應器，藉此控制帕耳帖熱電器件。巢套300可包含一串列埠350，其允許控制器308之微處理器經由介面310而與一外部主控制器154通信。另外，控制器308之微處理器可與電信號產生子系統304及熱控制子系統306通信(例如，經由一Plink工具(圖中未展示))。因此，經由控制器308、介面310及串列埠350之組合，電信號產生子系統304及熱控制子系統306可與外部主控制器154通信。依此方式，主控制器154可尤其藉由對輸出電壓調整執行縮放計算而輔助電信號產生子系統304。經由耦合至外部主控制器154之一顯示器件170而提供之一圖形使用者介面(GUI)(圖中未展示)可經構形以繪製分別自熱控制子系統306及電信號產生子系統304獲得之溫度及波形資料。替代地或另外，該GUI可允許更新控制器308、熱控制子系統306及電信號產生子系統304。如上文所討論，系統150可包含一成像器件194。在一些實施例中，成像器件194包括一光調變子系統422。光調變子系統422可包含一數位鏡面器件(DMD)或一微快門陣列系統(MSA)，其等之任一者可經構形以自一光源420接收光且將該接收光之一子集傳輸至顯微鏡450之一光學元件串中。替代地，光調變子系統422可包含自身產生光(且因此無需一光源420)之一器件，諸如一有機發光二極體顯示器(OLED)、一矽上液晶(LCOS)器件、一矽上鐵電液晶器件(FLCOS)或一透射式液晶顯示器(LCD)。光調變子系統422可為(例如)一投射器。因此，光調變子系統422能夠發射結構光及非結構光兩者。一適合光調變子系統422之一實例係來自Andor Technologies™之Mosaic™系統。在某些實施例中，系統150之成像模組164及/或動力模組162可控制光調變子系統422。在某些實施例中，成像器件194進一步包括一顯微鏡450。在此等實施例中，巢套300及光調變子系統422可個別地經構形以安裝於顯微鏡450上。顯微鏡450可為(例如)一標準研究級光學顯微鏡或螢光顯微鏡。因此，巢套300可經構形以安裝於顯微鏡450之置物台426上及/或光調變子系統422可經構形以安裝於顯微鏡450之一端口上。在其他實施例中，本文中所描述之巢套300及光調變子系統422可為顯微鏡450之整合組件。在某些實施例中，顯微鏡450可進一步包含一或多個偵測器440。在一些實施例中，偵測器440由成像模組164控制。偵測器440可包含一目鏡、一電荷耦合器件(CCD)、一攝影機(例如一數位攝影機)或其等之任何組合。若存在至少兩個偵測器440，則一偵測器可為(例如)一快圖框速率攝影機且另一偵測器可為一高敏感度攝影機。此外，顯微鏡450可包含一光學元件串，其經構形以自微流體器件360接收反射光及/或發射光且將該反射光及/或發射光之至少一部分聚焦於一或多個偵測器440上。顯微鏡之該光學元件串亦可包含用於不同偵測器之不同鏡筒透鏡(圖中未展示)，使得各偵測器上之最終放大倍數可不同。在某些實施例中，成像器件194經構形以使用至少兩個光源。例如，一第一光源420可用以產生結構光(例如，經由光調變子系統422)且一第二光源430可用以提供非結構光。第一光源420可產生用於光學致動電動及/或螢光激發之結構光，且第二光源430可用以提供亮場照明。在此等實施例中，動力模組164可用以控制第一光源420且成像模組164可用以控制第二光源430。顯微鏡450之光學元件串可經構形以：(1)自光調變子系統422接收結構光且當一微流體器件(諸如一光學致動之電動器件)由巢套300保持時將該結構光聚焦於該器件中之至少一第一區域上；及(2)自該微流體器件接收反射光及/或發射光且將此反射光及/或發射光之至少一部分聚焦至偵測器440上。光學元件串可進一步經構形以自一第二光源接收非結構光且當該微流體器件由巢套300保持時將該非結構光聚焦於該器件之至少一第二區域上。在某些實施例中，該微流體器件之該第一區域及該第二區域可為重疊區域。例如，該第一區域可為該第二區域之一子集。圖3B中展示將光供應至一光調變子系統422之第一光源420，光調變子系統422將結構光提供至系統450之顯微鏡450之光學元件串。圖中展示經由一分光器424而將非結構提供至光學元件串之第二光源430。來自光調變子系統422之結構光及來自第二光源430之非結構光一起自分光器424行進穿過光學元件串而到達一第二分光器424 (或二向色濾光器448，取決於由光調變子系統422提供之光)，其中該光經反射以向下穿過物鏡454而到達樣本平面428。接著，來自樣本平面428之反射光及/或發射光向上反向行進穿過物鏡454，穿過分光器及/或二向色濾光器448而到達二向色濾光器452。到達二向色濾光器452之光之僅一部分穿過且到達偵測器440。在一些實施例中，第二光源430發射藍光。使用一適當二向色濾光器452，自樣本平面428反射之藍光能夠穿過二向色濾光器452而到達偵測器440。相比而言，來自光調變子系統422之結構光自樣本平面428反射，但不穿過二向色濾光器452。在此實例中，二向色濾光器452濾除具有長於495奈米之一波長之可見光。自光調變子系統422濾除該光將僅在自光調變子系統發射之光不包含短於495奈米之任何波長時完成(如圖中所展示)。實際上，若來自光調變子系統422之光包含短於495奈米之波長(例如藍光波長)，則來自光調變子系統之一些光將穿過濾光器452而到達偵測器440。在此一實施例中，濾光器452用以改變自第一光源420及第二光源430到達偵測器440之光量之間的平衡。若第一光源420顯著地強於第二光源430，此可為有益的。在其他實施例中，第二光源430可發射紅光，且二向色濾光器452可濾除除紅光之外之可見光(例如具有短於650奈米之一波長之可見光)。 用於維持微流體器件之生長室內之細胞之存活性之額外系統組件為促進細胞群體之生長及/或擴增，有利於維持功能細胞之環境條件可由系統之額外組件提供。例如，此等額外組件可提供營養素、細胞生長傳信物種、pH調變、氣體交換、溫度控制及自細胞移除廢棄物。 微流體器件之調節表面在一些實施例中，微流體器件之至少一表面經調節以支援細胞生長、存活性、可移植性或其等之任何組合。在一些實施例中，調節實質上全部內表面。一調節表面可為促進微流體器件內之成功細胞培育之元件之一者。識別一適當調節表面需要平衡若干操作要求。第一，調節表面可提供用以使細胞免遭可用於製造此類微流體器件之材料類型影響之一接觸表面。在不受理論限制之情況下，調節表面可由水合水包圍，該水合水提供與細胞之一水接觸層而非一金屬接觸層。第二，調節表面可提供可在培育期間使用其來適當支撐至少一生物細胞之一接觸表面，其不會實質上抑制在培育完成之後自生長室移除細胞之能力。例如，諸多細胞需要具有一定程度之親水性之一接觸表面來充分貼附以可存活及/或生長。替代地，一些細胞需要具有一定程度之疏水性之一接觸表面來生長且呈現存活性之所要位準。另外，一些細胞需要選定蛋白質或肽基元存在於接觸表面內以啟始存活性/生長回應。第三，調節至少一表面可容許用於微流體器件中之動力實質上在正常作用電力範圍內起作用。例如，若採用光致動之動力，則調節表面可實質上容許光穿過調節表面，使得光致動之動力實質上不受抑制。至少一調節表面可包含生長室之一表面或流動區域之一表面或其等之一組合。在一些實施例中，複數個生長室之各者具有至少一調節表面。在其他實施例中，複數個流動區域之各者具有至少一調節表面。在一些實施例中，複數個生長室之各者及複數個流動區域之各者之至少一表面係調節表面。 包含一聚合物之調節表面至少一調節表面可包含一聚合物。該聚合物可共價鍵聯或非共價鍵聯至至少一表面。聚合物可具有各種結構基元，其包含嵌段聚合物(及共聚物)、星狀聚合物(星狀共聚物)、及接枝或梳形聚合物(接枝共聚物)，其等之全部可適合用於本文中。聚合物可包含具有伸烷基醚部分之一聚合物。各種含有伸烷基醚之聚合物可適合用於本文中所描述之微流體器件中。含有伸烷基醚之聚合物之一非限制性例示性類別係兩親性非離子嵌段共聚物，其包含在聚合物鏈內具有不同比率且位於不同位置中之聚環氧乙烷(PEO)亞單元及聚環氧丙烷(PPO)亞單元之嵌段。Pluronic®聚合物(BASF)係此類型之嵌段共聚物且在此項技術中被視為適合用在與活細胞接觸時。聚合物之平均分子量M _w在自約2000Da至約20KDa之範圍內。在一些實施例中，PEO-PPO嵌段共聚物可具有大於約10之一親水親油平衡值(HLB)(例如12至18)。用於產生一調節表面之特定Pluronic®聚合物包含Pluronic® L44、L64、P85及F127 (其包含F127NF)。含有伸烷基醚之聚合物之另一類別係聚乙二醇(PEG, Mw＜100,000Da)或替代地聚環氧乙烷(PEO, M _w＞100,000)。在一些實施例中，一PEG可具有約1000Da、約5000Da、約10,000Da或約20,000Da之一M _w。在其他實施例中，聚合物調節表面可包含含有羧酸部分之一聚合物。羧酸亞單元可為含有烷基部分、烯基部分或芳香族部分之一亞單元。一非限制性實例係聚乳酸(PLA)。在一些其他實施例中，聚合物調節表面可包含含有胺基甲酸酯部分之一聚合物，諸如(但不限於)聚胺基甲酸酯。在其他實施例中，聚合物調節表面可包含含有磺酸部分之一聚合物。磺酸亞單元可為含有烷基部分、烯基部分或芳香族部分之一亞單元。一非限制性實例係聚苯乙烯磺酸(PSSA)或聚茴香腦磺酸。此等後一例示性聚合物係聚電解質且可更改表面之特性以輔助/阻止黏著。在其他實施例中，聚合物調節表面可包含一聚合物，其在聚合物主鏈之一末端或來自該聚合物之主鏈之側鏈處含有磷酸部分。在其他實施例中，聚合物調節表面可包含含有糖類部分之一聚合物。在一非限制性實例中，多糖(諸如衍生自藻類或真菌多糖(諸如黃原膠或聚葡萄糖))可適合用於形成可輔助或防止細胞黏著之一聚合物調節表面。例如，具有約3Kda之一大小之聚葡萄糖聚合物可用以提供一微流體器件內之一調節表面。在其他實施例中，聚合物調節表面可包含含有核苷酸部分(即，核酸)之一聚合物，其可具有核糖核苷酸部分或脫氧核糖核苷酸部分。核酸可僅含有天然核苷酸部分或可含有非天然核苷酸部分，其包括核鹼基、核糖或磷酸部分類似物，諸如(但不限於) 7-去氮腺嘌呤、戊醣、甲基膦酸酯或硫代磷酸酯部分。含有核酸之一聚合物可包含可輔助或防止黏著之聚電解質。在其他實施例中，聚合物調節表面可包含含有胺基酸部分之一聚合物。含有胺基酸部分之聚合物可包含含有天然胺基酸之一聚合物或含有非天然胺基酸之一聚合物，其等之任一者可包含肽、多肽或蛋白質。在一非限制性實例中，蛋白質可為牛血清白蛋白(BSA)。在一些實施例中，一細胞外間質(ECM)蛋白質可提供於調節表面內以用於促進細胞生長之最佳化細胞黏著。可包含於一調節表面中之一細胞基質蛋白質可包含(但不限於)一膠原蛋白、一彈性蛋白、一含RGD之鈦(例如一纖連蛋白)或一層黏連蛋白。在其他實施例中，生長因子、細胞激素、內分泌素或其他細胞傳信物種可提供於微流體器件之至少一調節表面內。在進一步實施例中，聚合物調節表面可包含具有胺基部分之一聚合物。多胺聚合物可包含一天然多胺聚合物或一合成多胺聚合物。天然多胺之實例包含精胺、亞精胺及腐胺。在一些實施例中，聚合物調節表面可包含含有下列之一者以上之一聚合物：伸烷基氧化物部分、羧酸部分、磺酸部分、磷酸部分、糖類部分、核苷酸部分或胺基酸部分。在其他實施例中，聚合物調節表面可包含各具有下列各者之一個以上聚合物之一混合物：伸烷基氧化物部分、羧酸部分、磺酸部分、磷酸部分、糖類部分、核苷酸部分及/或胺基酸部分，其等可獨立地或同時併入至調節表面中。 共價鍵聯 之 調節表面在一些實施例中，至少一調節表面包含經構形以支援微流體器件內之一或多個生物細胞之細胞生長、存活性、可移植性或其等之任何組合之一共價鍵聯部分。該共價鍵聯部分可包含一鍵聯基團，其中該鍵聯基團共價鍵聯至微流體器件之一表面。該鍵聯基團亦鍵聯至經構形以支援微流體器件內之一或多個生物細胞之細胞生長、存活性、可移植性或其等之任何組合之該部分。該鍵聯基團鍵聯至其之該表面可包含微流體器件之基板之一表面，針對其中微流體器件包含一DEP構形之實施例，該表面可包含矽及/或二氧化矽。在一些實施例中，該共價鍵聯調節表面包含微流體器件之全部內表面。圖9中展示具有一調節表面之一微流體器件之一示意圖。如圖9中所見，一微流體器件900具有面向該微流體器件之一封閉區域902之一第一DEP基板904及一第二DEP基板906，微流體器件900可包含至少一生長室及/或流動區域。器件900可依其他方式構形，如同微流體器件100、200、240、290、400、500A至500E或600之任何者。封閉區域902可為其中維持生物細胞或將生物細胞輸入至其中或自其輸出生物細胞之區域。(第二DEP基板906之)內表面910及(第一DEP基板904之)內表面912經一調節表面916改質，調節表面916可為支援細胞生長、存活性、可移植性或其等之任何組合之任何部分。在此實施例中，調節表面經由矽烷氧基鍵聯基團914而共價鍵聯至內表面之氧化官能基。在一些實施例中，經構形以支援細胞生長、存活性、可移植性或其等之任何組合之共價鍵聯部分可包含：烷基或氟烷基(其包含全氟烷基)部分；單糖或多糖(其可包含(但不限於)聚葡萄糖)；醇類(其包含(但不限於)炔丙醇)；多元醇，其包含(但不限於)聚乙烯醇；伸烷基醚，其包含(但不限於)聚乙二醇；聚電解質(其包含(但不限於)聚丙烯酸或聚乙烯膦酸)；胺基(其包含其衍生物，諸如(但不限於)烷基化胺、羥烷基化胺基、胍鹽及含有一未芳香化氮環原子之雜環基，諸如(但不限於)嗎啉基或哌嗪基)；羧酸，其包含(但不限於)丙炔酸(其可提供羧酸陰離子表面)；膦酸，其包含(但不限於)乙炔基膦酸(其可提供膦酸陰離子表面)；磺酸陰離子；羧基甜菜鹼；磺基甜菜鹼；胺磺酸；或胺基酸。經構形以支援微流體器件內之一或多個生物細胞之細胞生長、存活性、可移植性或其等之任何組合之共價鍵聯部分可為如本文中所描述之任何聚合物，且可包含一或多個聚合物，其含有伸烷基氧化物部分、羧酸部分、糖類部分、磺酸部分、磷酸部分、胺基酸部分、核酸部分或胺基部分。在其他實施例中，經構形以支援一或多個生物細胞之細胞生長、存活性、可移植性或其等之任何組合之共價鍵聯部分可包含非聚合部分，諸如烷基部分、氟烷基部分(其包含(但不限於)全氟烷基)、胺基酸部分、醇部分、胺基部分、羧酸部分、膦酸部分、磺酸部分、胺磺酸部分或糖類部分。在一些實施例中，共價鍵聯部分可為烷基。該烷基可包括形成一直鏈之碳原子(例如至少10個碳或至少14個、16個、18個、20個、22個或22個以上碳之一直鏈)。因此，該烷基可為一無支鏈烷基。在一些實施例中，該烷基可包含一經取代之烷基(例如，該烷基中之一些碳可經氟化或經全氟化)。該烷基可包括接合至未經取代碳之一直鏈之經取代(例如，經氟化或經全氟化)碳之一直鏈。例如，該烷基可包含接合至一第二鏈段(其可包含一未經取代烷基)之一第一鏈段(其可包含全氟烷基)。該第一鏈段及該第二鏈段可直接或間接接合(例如，藉由一醚鍵)。該烷基之該第一鏈段可定位於鍵聯基團之遠端處，且該烷基之該第二鏈段可定位於鍵聯基團之近端處。在其他實施例中，該烷基可包含一支鏈烷基且可進一步具有中斷該烷基之烷基主鏈之一或多個伸芳基。在一些實施例中，該烷基或氟化烷基之一支鏈部分或經伸芳基中斷部分定位於至表面之共價鍵之遠端處之一點處。在其他實施例中，共價鍵聯部分可包含至少一個胺基酸，其可包含一個以上胺基酸。共價鍵聯部分可包含肽或蛋白質。在一些實施例中，共價鍵聯部分可包含胺基酸，其可提供用於支援細胞生長、存活性、可移植性或其等之任何組合之一兩性離子表面。共價鍵聯部分可包含一或多個糖。該等共價鍵聯糖可為單糖、雙糖或多糖。該等共價鍵聯糖可經改質以引入容許偶合至或(詳細言之)附接至表面之一反應配對部分。例示性反應配對部分可包含醛基、炔基或鹵基部分。可使多糖依一隨機方式改質，其中糖類單體之各者可經改質或該多糖內之糖類單體之僅一部分經改質以提供可直接或間接偶合至一表面之一反應配對部分。一範例可包含可經由一無支鏈鍵聯子而間接偶合至一表面之聚葡萄糖多糖。共價鍵聯部分可包含一或多個胺基。該胺基可為一經取代之胺部分、胍部分、含氮雜環部分或雜芳基部分。含胺基部分可具有容許微流體器件內及(視情況)生長室內之環境之pH改質之結構。共價鍵聯部分可包含一或多個羧酸、膦酸、胺磺酸或磺酸部分。在一些實施例中，共價鍵聯部分可包含一或多個核酸部分，其可具有經設計以自微流體器件內之生物細胞捕獲核酸之個別核苷酸之一序列。該等捕獲核酸可具有與來自生物細胞之核酸互補且可藉由雜交而捕獲核酸之一核苷酸序列。調節表面可由僅一種部分組成或可包含一種以上不同部分。例如，氟烷基調節表面(其包含全氟烷基)可具有全部相同之複數個共價鍵聯部分，例如，具有至表面之相同共價附接且具有相同數目個氟亞甲基單元(其包括支援生長及/或存活性及/或可移植性之氟烷基部分)。替代地，調節表面可具有附接至表面之一種以上部分。例如，調節表面可包含具有指定數目個亞甲基或氟亞甲基單元之烷基或氟烷基且可進一步包含附接至具有一帶電部分之表面之另一組部分，該帶電部分附接至具有更大數目個亞甲基或氟亞甲基單元之烷基或氟烷基鏈。在一些實施例中，具有一種以上附接部分之調節表面可經設計使得具有較大數目個主鏈原子且因此具有自共價附接至表面之一較大長度之一第一組附接配位基可提供容量來使體積較大部分呈現於調節表面處，而具有空間需求較小之不同末端且具有較少主鏈原子之一第二組附接配位基可有助於使整個基板表面官能化以防止與一矽或氧化鋁基板本身之非所要黏著或接觸。在另一實例中，附接至表面之部分可提供一兩性離子表面以依一隨機方式將交變電荷呈現於該表面上。 調節表面性質在一些實施例中，共價鍵聯部分可在共價鍵聯至微流體器件之表面(例如一經DEP構形之基板表面)時形成一單層。在一些實施例中，由共價鍵聯部分形成之調節表面可具有小於10奈米(例如，小於5奈米或約1.5奈米至約3.0奈米)之一厚度。在其他實施例中，由共價鍵聯部分形成之調節表面可具有約10奈米至約50奈米之一厚度。在一些實施例中，調節表面無需一完美形成單層適當地運用於一DEP構形內之操作。在各種實施例中，微流體器件之(若干)調節表面可提供所要電性質。在不受理論限制之情況下，影響一調節表面之穩健性之一因數係本徵電荷捕集。不同表面調節材料可捕集會導致材料被擊穿之電子。調節表面中之缺陷會導致電荷捕集且進一步導致調節表面被擊穿。除調節表面之組合物之外，其他因數(諸如疏水性材料之實體厚度)亦可影響DEP力。各種因數可更改調節表面之實體厚度，諸如調節表面形成於基板上之方式(例如氣相沈積、液相沈積、旋轉塗佈、溢流及靜電塗佈)。可使用一橢偏計來量測調節表面之實體厚度及均勻性。除調節表面之電性質之外，調節表面亦可具有有益於與生物分子一起使用之性質。例如，含有氟化(或全氟化)碳鏈之一調節表面可提供比烷基封端之鏈減少表面污損量之一益處。如本文中所使用，表面污損係指無差別材料沈積於微流體器件之表面上之量，其可包含諸如蛋白質及其降解物、核酸及各自降解物及其類似者之生物材料之永久性或半永久性沈積。可用於DEP構形中之調節表面之各種性質包含於下表中。如表中可見，針對條目1至7 (其等全部為如本文中所描述之共價鍵聯調節表面)，藉由橢圓偏振術而量測之厚度始終薄於條目8 (藉由非共價旋轉塗佈而形成之一CYTOP表面)之厚度(N/A表示整個表中無資料可用)。吾人發現，污損更多取決於表面之化學性質而非形成之模式，此係因為氟化表面通常比烷(烴)調節表面更少污損。表 1.藉由相較於一非共價形成之CYTOP表面使一表面共價改質而製備之各種調節表面之性質 表面改質類型 表面改質劑之式 厚度污損 1. 烷基封端之矽氧烷 (C ₁₈) CH ₃-(CH ₂) ₁₇-Si-(OCH ₃) ₃ ~ 2 nm 比氟化層更多污損 2. 氟烷基封端之烷基矽氧烷 (C ₁₆F) CF ₃-(CF ₂) ₁₃-(CH ₂) ₂-Si-(OCH ₃) ₃ ~2 nm 比烷基封端之層更抗污損 3. 氟烷基封端之烷基矽氧烷 C ₁₀F CF ₃-(CF ₂) ₇-(CH ₂) ₂-Si-(OCH ₃) ₃ ~1 nm 比烷基封端之層更抗污損 4. 烷基封端之矽氧烷 (C ₂₂) CH ₃-(CH ₂) ₂₁-Si-(OCH ₂CH ₃) ₃ ~2-2.5 nm 比氟化層更多污損 5. 烷基封端之矽氧烷 (C ₁₆) CH ₃-(CH ₂) ₁₅-Si-(OCH ₃) ₃ N/A 比氟化層更多污損 6. 氟烷基封端之烷氧基烷基矽氧烷 C ₆FC ₁₃ CF ₃-(CF ₂) ₅-(CH ₂) ₂-O-(CH ₂) ₁₁-Si(OCH ₃) ₃ ~2 nm N/A 7. 烷基封端之膦酸酯 C ₁₈PA CH ₃-(CH ₂) ₁₇-P=O(OH) ₂ N/A 比氟化層更多污損 8. CYTOP 含氟聚合物 ^{1, 2} ~30 nm 比烷基封端之層更抗污損 1. CYTOP結構：

2. 旋轉塗佈，非共價。 至表面之鍵聯基 團形成調節表面之共價鍵聯部分經由一鍵聯基團而附接至表面。該鍵聯基團可為藉由使一含矽氧烷之試劑與基板表面(其可由矽或氧化鋁形成)之氧化物反應而形成之矽烷氧基鍵聯基團。在一些其他實施例中，該鍵聯基團可為藉由使一含膦酸之試劑與矽或鋁基板表面之氧化物反應而形成之膦酸酯。 多部分調節表面可藉由一表面調節試劑(其經構形以已含有提供調節表面之部分)(例如烷基矽氧烷試劑或一經氟取代之烷基矽氧烷試劑，其可包含全氟矽氧烷試劑)之反應而形成共價鍵聯調節表面，如下文將描述。替代地，可藉由將支援細胞生長、存活性、可移植性或其等之任何組合之部分偶合至一表面改質配位基(其本身共價鍵聯至表面)而形成調節表面。 一調節表面之結構及製備方法在一些實施例中，共價鍵聯至介電泳基板之表面之氧化物的一調節表面具有式1之一結構：

式1 調節表面可共價鍵聯至介電泳基板之表面之氧化物。介電泳基板可為矽或氧化鋁，且氧化物可作為基板之天然化學結構之部分而存在或可如下文將討論般被引入。調節表面可經由一鍵聯基團LG而附接至氧化物，鍵聯基團LG可為藉由使矽氧烷或膦酸基與氧化物反應而形成之矽烷氧基或膦酸酯基。經構形以支援細胞生長、存活性、可移植性或其等之任何組合之部分可包含：烷基或氟烷基(其包含全氟烷基)部分；單糖或多糖(其可包含(但不限於)聚葡萄糖)；醇類(其包含(但不限於)炔丙醇)；多元醇，其包含(但不限於)聚乙烯醇；伸烷基醚，其包含(但不限於)聚乙二醇；聚電解質(其包含(但不限於)聚丙烯酸或聚乙烯膦酸)；胺基(其包含其衍生物，諸如(但不限於)烷化胺基、羥烷基化胺基、胍鹽及含有一未芳香化氮環原子之雜環基，諸如(但不限於)嗎啉基或哌嗪基)；羧酸，其包含(但不限於)丙炔酸(其可提供羧酸陰離子表面)；膦酸，其包含(但不限於)乙炔基膦酸(其可提供膦酸陰離子表面)；磺酸陰離子；羧基甜菜鹼；磺基甜菜鹼；胺磺酸；或胺基酸。烷基或氟烷基部分可具有等於或大於10個碳之一主鏈長度。在一些實施例中，該烷基或氟烷基部分可具有約10個、約12個、約14個、約16個、約18個、約20個或約22個碳之一主鏈長度。鍵聯基團LG可直接或間接連接至提供支援微流體器件內之細胞生長、存活性、可移植性或其等之任何組合之部分。當鍵聯基團LG直接連接至該部分時，選用鍵聯子L不存在且n係0。當鍵聯基團LG間接連接至該部分時，鍵聯子L存在且n係1。鍵聯子L可具有一直鏈部分，其中該直鏈部分之一主鏈可包含選自矽原子、碳原子、氮原子、氧原子、硫原子及磷原子(經受此項技術中已知之化學接合限制)之任何組合之1個至200個非氫原子。在一些非限制性實例中，可使用選自由醚、胺基、羰基、醯胺基或膦酸酯基組成之群組之一或多個部分之任何組合來中斷鍵聯子L。另外，鍵聯子L可具有中斷鍵聯子之主鏈之一或多個伸芳基、伸雜芳基或雜環基。在一些實施例中，鍵聯子L之主鏈可包含10個至20個原子。在其他實施例中，鍵聯子L之主鏈可包含約5個原子至約200個原子、約10個原子至約80個原子、約10個原子至約50個原子或約10個原子至約40個原子。在一些實施例中，主鏈原子全部為碳原子。在其他實施例中，主鏈原子並非全部為碳，而是可包含經受此項技術中已知之化學接合限制之矽原子、碳原子、氮原子、氧原子、硫原子或磷原子之任何可行組合。 表面調節試劑當在一步驟程序中將經構形以支援細胞生長、存活性、可移植性或其等之任何組合且藉此提供調節表面之部分添加至基板之表面時，式6之一表面調節試劑可用以引入調節表面。表面調節試劑可具有式6之一結構：

。式6 在式6之表面調節試劑中，表面調節試劑可包含一鍵聯基團LG，其可為矽氧烷或膦酸基。鍵聯基團LG可直接或間接鍵聯至經構形以支援細胞生長、存活性、可移植性或其等之任何組合之部分。LG可經由連接至一鍵聯子L之一第一端而直接(n=0)或間接(n=1)鍵聯至經構形以支援細胞生長、存活性、可移植性或其等之任何組合之部分。鍵聯子L可進一步包含一直鏈部分，其中該直鏈部分之一主鏈可具有選自矽原子、碳原子、氮原子、氧原子、硫原子及磷原子(經受此項技術中已知之化學接合限制)之任何組合之1個至200個非氫原子。該直鏈部分之主鏈可進一步包含一或多個伸芳基部分。經構形以支援細胞生長、存活性、可移植性或其等之任何組合之部分(「部分」)可包含：烷基或氟烷基(其包含全氟烷基)部分；單糖或多糖(其可包含(但不限於)聚葡萄糖)；醇類(其包含(但不限於)炔丙醇)；多元醇，其包含(但不限於)聚乙烯醇；伸烷基醚，其包含(但不限於)聚乙二醇；聚電解質(其包含(但不限於)聚丙烯酸或聚乙烯膦酸)；胺基(其包含其衍生物，諸如(但不限於)烷化胺基、羥烷基化胺基、胍鹽及含有一未芳香化氮環原子之雜環基，諸如(但不限於)嗎啉基或哌嗪基)；羧酸，其包含(但不限於)丙炔酸(其可提供羧酸陰離子表面)；膦酸，其包含(但不限於)乙炔基膦酸(其可提供膦酸陰離子表面)；磺酸陰離子；羧基甜菜鹼；磺基甜菜鹼；胺磺酸；或胺基酸。經構形以支援細胞生長、存活性、可移植性或其等之任何組合之部分可包含烷基或全氟烷基部分。該等烷基或全氟烷基部分可具有大於10個碳之一主鏈長度。經構形以支援細胞生長、存活性、可移植性或其等之任何組合之表面調節試劑之部分可包含糖類部分，且可為聚葡萄糖。在其他實施例中，經構形以支援細胞生長、存活性、可移植性或其等之任何組合之表面調節試劑之部分可包含伸烷基醚部分。該等伸烷基醚部分可為聚乙二醇。表面調節試劑可進一步包含一可裂解部分，其可定位於鍵聯子L內或可為經構形以支援細胞生長、存活性、可移植性或其等之任何組合之表面調節試劑之部分之部分。該可裂解部分可經構形以容許破壞調節表面，藉此促進一或多個生物細胞之可移植性。在一些實施例中，可在一多步驟程序中將支援細胞生長、存活性、可移植性或其等之任何組合之部分添加至基板之表面。當部分依一逐步方式偶合至表面時，鍵聯子L可進一步包含一偶合基團CG，如式2中所展示。

式2 在一些實施例中，偶合基團CG表示來自一反應部分R _x與經構形以與一反應配對部分R _px反應之一部分之反應之所得部分。例如，一典型CG可包含甲醯胺基，其係胺基與羧酸之一衍生物(諸如活性酯、醯氯或其類似者)之反應結果。CG可包含可在一反應部分與其各自反應配對部分反應之後形成之伸三唑基、甲醯胺基、硫代醯胺基、肟、烷硫基、二硫化物、醚或烯基或任何其他適合基團。偶合基團CG可定位於其中附接部分之鍵聯子L之第二端處。在一些其他實施例中，偶合基團CG可中斷鍵聯子L之主鏈。在一些實施例中，偶合基團CG係伸三唑基，其係炔基與疊氮基之間的一反應之結果，炔基及疊氮基之任一者可為此項技術中已知用於點擊偶合反應中之反應部分或反應配對部分。亦可進一步取代伸三唑基。例如，二苯并環辛烯基稠合伸三唑基部分可源於具有二苯并環辛炔基反應配對部分R _px之一調節改質劑與表面改質配位基之疊氮基反應部分R _x之反應，如下列段落中將更詳細描述。各種二苯并環辛炔基改質分子在此項技術中係已知的或可經合成以併入經構形以支援細胞生長、存活性、可移植性或其等之任何組合之一部分。當在一多步驟程序中形成調節表面時，可藉由一調節改質劑(式5)與一基板(其具有共價鍵聯至其之一表面改質配位基)之反應而引入支援細胞生長、存活性、可移植性或其等之任何組合之部分，該表面改質配位基具有式3之一結構。

式5 式3 式3之中間改質表面具有附接至其之一表面改質配位基，其具有-LG-(L'') _j-R _x之一式，該式鍵聯至基板之氧化物，且如上文針對式1之調節表面所描述般類似形成。DEP基板之表面係如上文所描述，且包含基板固有或引入至其內之氧化物。鍵聯基團LG係如上文所描述。一鍵聯子L''可存在(j=1)或不存在(j=0)。鍵聯子L''可具有一直鏈部分，其中該直鏈部分之一主鏈可包含選自矽原子、碳原子、氮原子、氧原子、硫原子及磷原子(經受此項技術中已知之化學接合限制)之任何組合之1個至100個非氫原子。在一些非限制性實例中，可使用醚、胺基、羰基、醯胺基或膦酸酯基之任何組合來中斷鍵聯子L''。另外，鍵聯子L''可具有中斷鍵聯子之主鏈之一或多個伸芳基、伸雜芳基或雜環基。在一些實施例中，鍵聯子L''之主鏈可包含10個至20個原子。在其他實施例中，鍵聯子L''之主鏈可包含約5個原子至約100個原子、約10個原子至約80個原子、約10個原子至約50個原子或約10個原子至約40個原子。在一些實施例中，主鏈原子全部為碳原子。在其他實施例中，主鏈原子並非全部為碳，而是可包含經受此項技術中已知之化學接合限制之矽原子、碳原子、氮原子、氧原子、硫原子或磷原子之任何可行組合。一反應部分R _x存在於表面改質配位基與表面之共價鍵遠端處之表面改質配位基之末端處。反應部分R _x係用於偶合反應以引入支援細胞生長、存活性、可移植性或其等之任何組合之部分之任何適合反應部分。在一些實施例中，反應部分R _x可為疊氮基、胺基、溴基、硫醇基、活性酯基、琥珀醯亞胺基或炔基部分。 調節改質劑調節改質劑(式5)經構形以供應支援細胞生長、存活性、可移植性或其等之任何組合之部分。

式5 經構形以支援細胞生長、存活性、可移植性或其等之任何組合之調節改質劑之部分藉由一反應配對部分R _px與反應部分R _x之反應而鍵聯至表面改質配位基。反應配對部分R _px係經構形以與各自反應部分R _x反應之任何適合反應部分。在一非限制性實例中，一適合反應配對部分R _px可為炔基且反應部分R _x可為疊氮基。替代地，反應配對部分R _px可為疊氮基部分且各自反應部分R _x可為炔基。在其他實施例中，反應配對部分R _px可為活性酯官能基且反應部分R _x可為胺基。在其他實施例中，反應配對部分R _px可為醛基且反應部分R _x可為胺基。其他反應部分-反應配對部分組合係可行的，且此等實例絕不具限制性。經構形以支援細胞生長、存活性、可移植性或其等之任何組合之式5之調節改質劑之部分可包含：烷基或氟烷基(其包含全氟烷基)部分；單糖或多糖(其可包含(但不限於)聚葡萄糖)；醇類(其包含(但不限於)炔丙醇)；多元醇，其包含(但不限於)聚乙烯醇；伸烷基醚，其包含(但不限於)聚乙二醇；聚電解質(其包含(但不限於)聚丙烯酸或聚乙烯膦酸)；胺基(其包含其衍生物，諸如(但不限於)烷化胺基、羥烷基化胺基、胍鹽及含有一未芳香化氮環原子之雜環基，諸如(但不限於)嗎啉基或哌嗪基)；羧酸，其包含(但不限於)丙炔酸(其可提供羧酸陰離子表面)；膦酸，其包含(但不限於)乙炔基膦酸(其可提供膦酸陰離子表面)；磺酸陰離子；羧基甜菜鹼；磺基甜菜鹼；胺磺酸；或胺基酸。提供增強細胞生長、存活性、可移植性或其等之任何組合之式5之調節改質劑之部分可直接(L'，其中m=0)或間接連接至反應配對部分R _px。當反應配對部分R _px間接連接至提供增強細胞生長、存活性、可移植性或其等之任何組合之部分時，反應配對部分R _px可連接至一鍵聯子L' (m=1)。反應配對部分R _px可連接至鍵聯子L'之一第一端，且提供增強細胞生長、存活性、可移植性或其等之任何組合之部分可連接至鍵聯子L'之一第二端。鍵聯子L'可具有一直鏈部分，其中該直鏈部分之一主鏈可包含選自矽原子、碳原子、氮原子、氧原子、硫原子及磷原子(經受此項技術中已知之化學接合限制)之任何組合之1個至100個非氫原子。在一些非限制性實例中，可使用醚、胺基、羰基、醯胺基或膦酸酯基之任何組合來中斷鍵聯子L'。另外，鍵聯子L'可具有中斷鍵聯子L'之主鏈之一或多個伸芳基、伸雜芳基或雜環基。在一些實施例中，鍵聯子L'之主鏈可包含10個至20個原子。在其他實施例中，鍵聯子L'之主鏈可包含約5個原子至約100個原子、約10個原子至約80個原子、約10個原子至約50個原子或約10個原子至約40個原子。在一些實施例中，主鏈原子全部為碳原子。在其他實施例中，主鏈原子並非全部為碳，而是可包含經受此項技術中已知之化學接合限制之矽原子、碳原子、氮原子、氧原子、硫原子或磷原子之任何可行組合。當調節改質劑(式5)與具有一表面改質配位基(式3)之表面反應時，形成具有式2之一調節表面之一基板。接著，鍵聯子L'及鍵聯子L''形式上係鍵聯子L之部分，且反應配對部分R _px與反應部分R _x之反應產生式2之偶合基團CG。 表面改質劑表面改質劑係具有LG-(L'') _j-R _x(式4)之一結構之一化合物。鍵聯基團LG共價鍵聯至介電泳基板之表面之氧化物。介電泳基板可為矽或氧化鋁，且氧化物可作為基板之天然化學結構之部分而存在或可如本文中所討論般被引入。鍵聯基團LG可為由矽氧烷或膦酸基與基板之表面上之氧化物之反應形成之矽烷氧基或膦酸酯基。反應部分R _x係如上文所描述。反應部分R _x可直接(L''，j=0)或經由一鍵聯子L''(j=1)而間接連接至鍵聯基團LG。鍵聯基團LG可附接至鍵聯子L''之一第一端且反應部分R _x可連接至鍵聯子L''之一第二端，在表面改質配位基已如式3般附接至表面之後，該第二端將位於基板之表面之遠端處。

式4 式3 鍵聯子L''可具有一直鏈部分，其中該直鏈部分之一主鏈包含選自矽原子、碳原子、氮原子、氧原子、硫原子及磷原子之任何組合之1個至100個非氫原子。在一些非限制性實例中，可使用醚、胺基、羰基、醯胺基或膦酸酯基之任何組合來中斷鍵聯子L''。另外，鍵聯子L''可具有中斷鍵聯子L''之主鏈之一或多個伸芳基、伸雜芳基或雜環基。在一些實施例中，鍵聯子L''之主鏈可包含10個至20個原子。在其他實施例中，鍵聯子L''之主鏈可包含約5個原子至約100個原子、約10個原子至約80個原子、約10個原子至約50個原子或約10個原子至約40個原子。在一些實施例中，主鏈原子全部為碳原子。在其他實施例中，主鏈原子並非全部為碳，而是可包含經受此項技術中已知之化學接合限制之矽原子、碳原子、氮原子、氧原子、硫原子或磷原子之任何可行組合。 可裂解部分在各種實施例中，下列之任何者可進一步包含一可裂解部分(如下文將討論)：支援細胞生長、存活性、可移植性或其等之任何組合之部分、鍵聯子L、鍵聯子L'、鍵聯子L''或偶合基團CG。該可裂解部分可經構形以容許破壞一微流體器件之一調節表面，其促進一或多個生物細胞之可移植性。在一些實施例中，可期望一或多個生物細胞之可移植性能夠在培養細胞一時間週期之後移動細胞且尤其能夠自微流體器件輸出細胞。 基板之組合物據此，提供一組合物，其包含：一基板，其具有一介電泳(DEP)構形及一表面；及一調節表面，其共價鍵聯至該基板之該表面之氧化物部分。該基板上之該調節表面可具有式1或式2之一結構：

；式1 式2 其中：LG係一鍵聯基團；L係一鍵聯子，其可存在(n=1)或不存在(n=0)；部分係支援微流體器件內之細胞生長、存活性、可移植性或其等之任何組合之部分；及CG係一偶合基團，如本文中所界定。調節表面可包含共價鍵聯至表面之氧化物部分之一鍵聯基團LG。該鍵聯基團可進一步鍵聯至經構形以支援細胞生長、存活性、可移植性或其等之任何組合之一部分。該鍵聯基團可為矽烷氧基鍵聯基團。在其他實施例中，該鍵聯基團可為膦酸酯基。該鍵聯基團可直接或間接鍵聯至經構形以支援細胞生長、存活性、可移植性或其等之任何組合之該部分。該鍵聯基團可經由連接至一鍵聯子之一第一端而間接鍵聯至經構形以支援細胞生長、存活性、可移植性或其等之任何組合之該部分。該鍵聯子可進一步包含一直鏈部分，其中該直鏈部分之一主鏈可具有選自矽原子、碳原子、氮原子、氧原子、硫原子及磷原子之任何組合之1個至200個非氫原子，如上文所討論。該直鏈部分之主鏈可進一步包含一或多個伸芳基部分。鍵聯子可具有如上文所界定之一偶合基團CG。偶合基團CG可包含伸三唑基部分。該伸三唑基部分可中斷鍵聯子之直鏈部分或可連接至鍵聯子之直鏈部分之一第二端。鍵聯子之第二端可位於基板之表面之遠端處。經構形以支援細胞生長、存活性、可移植性或其等之任何組合之部分可包含：烷基或氟烷基(其包含全氟烷基)部分；單糖或多糖(其可包含(但不限於)聚葡萄糖)；醇類(其包含(但不限於)炔丙醇)；多元醇，其包含(但不限於)聚乙烯醇；伸烷基醚，其包含(但不限於)聚乙二醇；聚電解質(其包含(但不限於)聚丙烯酸或聚乙烯膦酸)；胺基(其包含其衍生物，諸如(但不限於)烷化胺基、羥烷基化胺基、胍鹽及含有一未芳香化氮環原子之雜環基，諸如(但不限於)嗎啉基或哌嗪基)；羧酸，其包含(但不限於)丙炔酸(其可提供羧酸陰離子表面)；膦酸，其包含(但不限於)乙炔基膦酸(其可提供膦酸陰離子表面)；磺酸陰離子；羧基甜菜鹼；磺基甜菜鹼；胺磺酸；或胺基酸。在一些實施例中，將不同部分之一混合物(諸如(但不限於)提供一兩性離子調節表面之陰離子官能基及陽離子官能基之一混合物)併入於調節表面中。調節表面可包含烷基或全氟烷基部分。該等烷基或全氟烷基部分可具有大於10個碳之一主鏈長度。調節表面可包含糖類部分，且可為聚葡萄糖。在其他實施例中，調節表面可包含伸烷基醚部分。該等伸烷基醚部分可為聚乙二醇。調節表面可進一步包含一可裂解部分。該可裂解部分可經構形以容許破壞調節表面，藉此促進一或多個生物細胞之可移植性。提供另一組合物，其包含：一基板，其包含一介電泳(DEP)構形及一表面；及一表面改質配位基，其共價鍵聯至該基板之該表面之氧化物部分。具有一表面改質配位基之該基板可具有式3之一結構：

式3 其中：LG係鍵聯基團；L''係一選用鍵聯子，j係0或1。鍵聯子L''當j=1時，鍵聯子L''存在，且當j=0時，鍵聯子L''不存在；及R _x係如本文中所描述之一反應部分。表面改質配位基之反應部分可為疊氮基、胺基、溴基、硫醇基、活性酯基、琥珀醯亞胺基或炔基部分。表面改質配位基可經由一鍵聯基團而共價鍵聯至氧化物部分。該鍵聯基團可為矽烷氧基部分。在其他實施例中，該鍵聯基團可為膦酸酯基。該鍵聯基團可經由一鍵聯子而間接連接至表面改質配位基之反應部分。該鍵聯基團可附接至該鍵聯子之一第一端且該反應部分可附接至該鍵聯子之一第二端。鍵聯子L''可包含一直鏈部分，其中該直鏈部分之一主鏈包含選自矽原子、碳原子、氮原子、氧原子、硫原子及磷原子之任何組合之1個至100個非氫原子。鍵聯子L''之主鏈可包含10個至20個原子。在其他實施例中，鍵聯子L''之主鏈可包含約5個原子至約50個原子。在一些實施例中，鍵聯子L''之主鏈可全部為碳原子。該直鏈部分之主鏈可包含一或多個伸芳基部分。鍵聯子L''可包含伸三唑基部分。該伸三唑基部分可中斷鍵聯子L''或可附接於鍵聯子L''之一末端處。表面改質配位基可包含一可裂解部分。該可裂解部分可經構形以容許破壞一微流體器件之一調節表面，藉此促進一或多個生物細胞之可移植性。 製備之方法在一些實施例中，使用化學氣相沈積來將調節表面或表面改質配位基沈積於微流體器件之內表面上。調節表面/表面改質配位基可透過分子之氣相沈積而達成緊密堆積單層，其中包括調節表面/表面改質配位基之分子共價鍵聯至微流體器件(100、200、240、290、400、500A至500E、600)之任何者之內表面之分子。為達成一所要堆積密度，可在至少110°C之一溫度(例如至少120°C、130°C、140°C、150°C、160°C等等)處氣相沈積包括(例如)烷基封端之矽氧烷之分子達至少15小時之一週期(例如至少20、25、30、35、40、45或45以上小時)。通常在真空下且在存在一水源時執行此氣相沈積，諸如一含水硫酸鹽(例如MgSO ₄·7H ₂0)。通常，氣相沈積之溫度升高及持續時間增加產生調節表面/表面改質配位基之改良特性。在一些實施例中，可藉由一液相中之反應而引入調節表面或表面改質配位基。為製備微流體表面，可藉由一氧氣電漿處理而處理蓋、微流體線路材料及電極活性基板，該氧氣電漿處理可移除各種雜質，同時引入氧化表面(例如可如本文中所描述般共價改質之表面處之氧化物)。可(例如)在真空條件下依100 W操作氧氣電漿清洗器達60秒。替代地，可使用液相處理，其包含用以氧化表面之氧化劑，諸如過氧化氫。例如，鹽酸及過氧化氫之一混合物或硫酸及過氧化氫之一混合物(例如食人魚溶液，其可具有自約3:1至約7:1範圍內之硫酸與過氧化氫之一比率)。例如，可藉由預清洗蓋、微流體線路材料及電極活性基板而視情況改良氣相沈積程序。例如，此預清洗可包含一溶劑浴，諸如丙酮浴、乙醇浴或其等之一組合。該溶劑浴可包含音波處理。在一些實施例中，在已組裝微流體器件以形成界定一微流體線路之一封閉體之後使用氣相沈積來塗佈微流體器件之(若干)內表面。當具有一表面改質配位基之一基板進一步與調節改質劑反應以製備具有一調節表面之基板時，可就地使用任何適合溶劑來執行反應，該任何適合溶劑將溶解試劑且不會破壞微流體線路材料或具有一表面改質配位基之表面。在一些實施例中，溶劑係一水溶液。 製備一調節表面或包含一表面改質配位基之一表面之方法據此，提供製備具有一介電泳(DEP)構形之一微流體器件之一改質表面之一方法，其包含下列步驟：提供一微流體器件之一基板之一表面，其中該基板包含一DEP構形；使該表面之氧化物與一改質劑反應，藉此將該基板之該表面轉換成一改質表面。在一些實施例中，該基板之該表面可經電漿清洗以將氧化物提供於該表面上。在一些實施例中，可在組裝該微流體器件之前電漿清洗該表面。在其他實施例中，可在組裝該微流體器件之後電漿清洗該表面。藉由使表面暴露於包括改質劑之一液體而執行方法中之使表面之氧化物與改質劑反應之步驟。在一些實施例中，可藉由在減小壓力處使表面暴露於含有改質劑之一蒸汽而執行使表面之氧化物反應之步驟。在一些實施例中，改質劑可包含一表面調節試劑，其具有經構形以與表面共價反應之一第一部分及經構形以支援細胞生長、存活性、可移植性或其等之任何組合之一第二部分，藉此使表面改質為經調節以支援細胞生長、存活性、可移植性或其等之任何組合之一表面。表面調節試劑可具有式6之一結構：

式6 第一部分可包含一鍵聯基團LG，其可為矽氧烷或膦酸基。鍵聯基團LG可直接或間接鍵聯至經構形以支援細胞生長、存活性、可移植性或其等之任何組合之部分。第一部分可直接(n=0)或經由連接至一鍵聯子L之一第一端而間接(n=1)鍵聯至第二部分，該第二部分係經構形以支援細胞生長、存活性、可移植性或其等之任何組合之部分。鍵聯子L可進一步包含一直鏈部分，其中該直鏈部分之一主鏈可具有選自矽原子、碳原子、氮原子、氧原子、硫原子及磷原子之任何組合之1個至200個非氫原子。該直鏈部分之主鏈可進一步包含一或多個伸芳基部分。表面調節試劑之第二部分(「部分」)可包含：烷基或氟烷基(其包含全氟烷基)部分；單糖或多糖(其可包含(但不限於)聚葡萄糖)；醇類(其包含(但不限於)炔丙醇)；多元醇，其包含(但不限於)聚乙烯醇；伸烷基醚，其包含(但不限於)聚乙二醇；聚電解質(其包含(但不限於)聚丙烯酸或聚乙烯膦酸)；胺基(其包含其衍生物，諸如(但不限於)烷化胺基、羥烷基化胺基、胍鹽及含有一未芳香化氮環原子之雜環基，諸如(但不限於)嗎啉基或哌嗪基)；羧酸，其包含(但不限於)丙炔酸(其可提供羧酸陰離子表面)；膦酸，其包含(但不限於)乙炔基膦酸(其可提供膦酸陰離子表面)；磺酸陰離子；羧基甜菜鹼；磺基甜菜鹼；胺磺酸；或胺基酸。表面調節試劑之第二部分可包含烷基或全氟烷基部分。該等烷基或全氟烷基部分可具有大於10個碳之一主鏈長度。表面調節試劑之第二部分可包含糖類部分，且可為聚葡萄糖。在其他實施例中，表面調節試劑之第二部分可包含伸烷基醚部分。該等伸烷基醚部分可為聚乙二醇。表面調節試劑之第二部分可進一步包含一可裂解部分，其可定位於鍵聯子L內或可為表面調節試劑之第二部分之部分。該可裂解部分可經構形以容許破壞調節表面，藉此促進一或多個生物細胞之可移植性。在各種實施例中，改質劑可包含一表面改質劑，其具有如上文所界定之式4之一結構，其中該表面改質劑包含：一第一部分LG，其經構形以與表面反應；及一第二部分R _x，其可為一反應部分或可經改質以包含一反應部分(該反應部分包含疊氮基、胺基、溴基、硫醇基、活性酯基、琥珀醯亞胺基或炔基部分)，藉此將表面轉換成包括一表面改質配位基之一表面，該表面改質配位基具有式3之一結構，如上文所描述。在一些實施例中，該表面改質劑之該第一部分(其經構形以與表面之氧化物反應)可為矽氧烷或膦酸。在一些實施例中，該方法包含下列步驟：使包括一表面改質配位基(式3)之表面與一調節改質劑反應，該調節改質劑包含經構形以支援細胞生長、存活性、可移植性或其等之任何組合之一第一部分及經構形以與該表面改質配位基之反應部分反應之一第二部分R _px；藉此提供經構形以支援一生物細胞之細胞生長、存活性、可移植性或其等之任何組合之一表面，該表面具有式2之一結構，如上文所描述。該調節改質劑可具有式5之一結構。在一些實施例中，該調節改質劑之該第一部分包括下列之至少一者：環氧烷部分、胺基酸部分、糖類部分、陰離子部分、陽離子部分及兩性離子部分。在各種實施例中，表面調節試劑、表面改質劑或調節改質劑之任何者可進一步包含如本文中所描述之一可裂解部分。 含有其他組分之調節表面調節表面可包含除由一共價鍵聯部分形成之一聚合物或一調節表面之外之其他組分，或調節表面除包含由一共價鍵聯部分形成之一聚合物或一調節表面之外，亦可另外包含其他組分，該共價鍵聯部分包含生物相容金屬離子(例如鈣、鈉、鉀或鎂)、抗氧化劑、表面活性劑及/或必需營養素。一非限制性例示性列表包含：維生素，諸如B7、α-生育酚、α-生育酚乙酸酯、維生素A及其乙酸酯；蛋白質，諸如BSA、過氧化氫酶、胰島素、轉鐵蛋白、超氧化物歧化酶；小分子，諸如皮質固酮、D-半乳糖、乙醇胺鹽酸鹽、還原谷胱甘肽、L-肉鹼鹽酸鹽、亞油酸、亞麻酸、黃體固酮、腐胺二鹽酸鹽及三碘甲腺原氨酸；及鹽，其包含(但不限於)亞硒酸鈉、磷酸鈉、磷酸鉀、磷酸鈣及/或磷酸鎂。抗氧化劑可包含(但不限於)類胡蘿蔔素、肉桂酸及其衍生物、阿魏酸、多酚(諸如類黃酮、醌及其衍生物)(其包含米托蒽醌-Q)、N-乙醯半胱胺酸及抗氧化維生素(諸如抗壞血酸、維生素E及其類似者)。調節表面可包含一培養基補充劑(諸如B-27®補充劑)，其含有抗氧化劑及上文所列之其他組分之諸多者。B-27®補充劑係來自ThermoFisher Scientific (Cat# 17504044)之市售(50倍)無血清培養基。在一些實施例中，至少一調節表面可包含哺乳動物血清之一或多個組分。在一些實施例中，哺乳動物血清係胎牛血清(FBS)或小牛血清(FCS)。調節表面可包含哺乳動物血清之特定組分(諸如常見於血清中之特定數量及類型之蛋白質)，其之界定數量或類型可自無血清培養基或合成培養基提供。在其他實施例中，至少一調節表面不包含一哺乳動物血清。在各種實施例中，至少一調節表面可不包含任何鈦、鎳或鐵金屬離子。在其他實施例中，至少一調節表面可不包含任何顯著濃度之鈦、鎳或鐵金屬離子。在其他實施例中，至少一調節表面可不包含任何金、鋁或鎢金屬離子。 用以減弱黏著之試劑處理 / 試劑之混合物當在一微流體器件內培養細胞時，細胞主動分泌蛋白質及其他生物分子且被動地流出可黏著至該微流體器件內之表面之類似生物分子。培養細胞可彼此黏著或黏著至調節表面，且變得難以自生長室移除以自該微流體器件輸出。另外，在一些情況下，可期望將相同於或不同於培養細胞之額外細胞類型帶入至該微流體件中。此等新傳遞細胞亦可變得黏著至累積於微流體環境內之表面污損，且難以在一隨後時間點處自該器件移除。使用蛋白酶(諸如胰蛋白酶或Accutase® (具有蛋白質水解活性及溶膠原活性之一酶促混合物，Innovative Cell Technologies))之處理無法提供充分功效足以(針對一非限制性實例)容許黏著細胞自微流體器件輸出之效能。提供抗黏著性質之一或多個蛋白質及/或肽可用作用以減弱此兩種情形之黏著之一混合物。可使用具有抵抗各種細胞黏著機制之一者之活性之生物分子或小分子。可受抑制之一些細胞黏著機制可為活性肌動蛋白纖維形成及相關程序，其可藉由使用化合物(諸如細胞鬆弛素B (New England Biosciences Cat No: M0303S)、微纖維延伸之一小分子抑制劑)而達成。可藉由使用含RGD之肽而達成特定受體驅動黏著程序，諸如(但不限於)抑制整合素受體介導黏著至纖連蛋白(其可見於一污損表面上)。另一類型之污損材料(即，自死細胞釋放之核酸)可吸引細胞結合，其可藉由使用一核酸內切酶(其將裂解污損核酸)而達成。一特定核酸內切酶(去氧核糖核酸酶1 (DNase 1，Sigma Aldrich，Catalog No. AMPD1-1KT))亦結合至肌動蛋白以因此提供黏著之一雙活性阻斷。在一些實施例中，全部三個阻斷劑之一混合物可用以防止/減弱細胞黏著。 一般處理協定 / 在培養之後：針對已在一微流體器件內生長2天、3天、4天或4天以上之細胞，三個抗黏著劑之混合物或單一抗黏著劑(如下文將描述)可流動至該微流體器件中且被允許在輸出細胞之前之約20分鐘、約30分鐘、約40分鐘、約50分鐘或約60分鐘之一時間週期內擴散至生長室中。 預處理針對待輸入至一微流體器件中之細胞，可在含有混合物或單一抗黏著劑之一培養基中預培育細胞達約30分鐘，接著，將細胞輸入至微流體晶片。抑制在無需進一步添加試劑之情況下持續1小時、2小時、3小時或3小時以上之時間週期。 RGD三肽(mw. 614.6，Santa Cruz Biotechnology Cat No: sc-201176)可以約0.1毫莫耳至約20毫莫耳之一濃度存在於培養基或預輸入培育基中。在一些實施例中，RGD三肽可以約0.1毫莫耳、約0.5毫莫耳、約0.7毫莫耳、約1.0毫莫耳、約3.0毫莫耳、約5.0毫莫耳、約8.0毫莫耳、約10.0毫莫耳或該範圍內之任何值之一濃度存在。細胞鬆弛素B可以約0.01毫莫耳或約50毫莫耳或約0.01毫莫耳、約0.05毫莫耳、約0.1毫莫耳、約2毫莫耳、約4毫莫耳、約6毫莫耳、約8毫莫耳、約10毫莫耳、約20毫莫耳、約30毫莫耳、約50毫莫耳或該範圍內之任何值之一濃度存在於預輸入培育基中。DNase 1可以約0.001 U/微升至約10 U/微升或約0.001 U/微升、約0.005 U/微升、約0.01 U/微升、約0.05 U/微升、約1.0 U/微升、約5.0 U/微升、約10 U/微升或該範圍內之任何值之一濃度存在。在一些實施例中，一單一試劑可用以減弱已在一微流體器件中培養細胞之前或已在一微流體器件中培養細胞之後之黏著。例如，RGD三肽可以5毫克/毫升之一濃度而用於預培育或可在輸出之前作為一處理劑流入該微流體器件內。可使用之另一抑制劑係四肽纖連蛋白抑制劑(Arg-Gly-Asp-Ser-OH，mw. 433.4，Santa Cruz Biotechnology Cat No: sc-202156)。該纖連蛋白抑制劑可以約1.75微克/毫升(4微莫耳)之一濃度使用。類似於將蛋白質或小分子試劑用以減弱或防止黏著，細胞外黏著相關蛋白質之抗體可用以影響輸出及微流體器件內之可移植性。一非限制性實例係抗B1整合素：選殖M-106 (Santa Cruz Biotechnology Cat No: sc-8978)。 含有一可裂解部分之調節表面在一些實施例中，調節表面可具有併入於調節表面之共價或非共價鍵聯分子內之可裂解部分。調節表面可包含具有上述之一功能之肽基元(諸如RGD)，或其可具有促進細胞生長或提供細胞增殖之接觸線索之另一肽基元。在其他實施例中，調節表面對細胞提供非特定支援，且可僅用以緩衝來自微流體器件之矽或鋁氧化物表面之細胞。可期望在完成細胞培養之一週期之後破壞調節表面以促進微流體器件之一生長室內之擴增細胞群體之輸出。此可用於細胞展現黏著行為時。可破壞調節表面，藉由併入其他肽基元(其係並非由所關注之細胞高度分泌之一蛋白酶之酶基)而部分或全部移除調節表面。在一非限制性實例中，ENLYQS (Glu-Asn-Leu-Tyr-Gln-Ser)之肽基元可依預設計時間間隔併入至一調節表面中。此基元係高度序列特定且因此用於高度受控裂解之TEV蛋白酶(菸草蝕刻病毒半胱胺酸蛋白酶，Sigma Aldrich catalog no. T4455)之一酶基。在完成培養週期之後，TEV蛋白酶可流動至微流體器件中且被允許擴散至生長室之隔離區域中。接著，破壞調節表面以促進微流體器件內之細胞之輸出。因此，各種其他解蛋白基元可經設計且併入至一調節表面中以藉由熟習此項技術者可想出之一適合特定蛋白酶而裂解。 流體培養基關於與具有一通道及一或多個生長室之微流體器件有關之上述討論，一流體培養基(例如一第一培養基及/或一第二培養基)可為能夠使一細胞維持於一實質上可變狀態中之任何流體。該可變狀態將取決於生物微小物體及所執行之培養實驗。第一流體培養基及/或第二流體培養基可提供細胞存活所需之流體組分及溶解氣體組分兩者，且亦可使用緩衝流體培養基或pH監測或兩者來使pH維持於一所要範圍內。若該細胞係一哺乳動物細胞，則第一流體培養基及/或第二流體培養基可包含此項技術中已知之哺乳動物血清或一無血清合成培養基，其能夠提供必需營養素、內分泌素、生長因子或細胞生長信號。類似於上述調節表面，第一流體培養基及/或第二流體培養基可包含胎牛血清(FBS)或小牛血清(FCS)。替代地，第一流體培養基及/或第二流體培養基可不包含任何動物源血清，但可包含一合成培養基，其可包含生理相關金屬離子(其包含(但不限於)鈉、鉀、鈣、鎂及/或鋅)抗氧化劑、表面活性劑及/或必需營養素之任何者或全部。合成培養基可為無血清的，同時仍含有一些蛋白質，其中該等蛋白質具有經界定數量及類型。一無血清培養基中之組分之一非限制性例示性列表包含：維生素，諸如B7、α-生育酚、α-生育酚乙酸酯、維生素A及其乙酸酯；蛋白質，諸如BSA、過氧化氫酶、胰島素、轉鐵蛋白、超氧化物歧化酶；小分子，諸如皮質固酮、D-半乳糖、乙醇胺鹽酸鹽、還原谷胱甘肽、L-肉鹼鹽酸鹽、亞油酸、亞麻酸、黃體固酮、腐胺二鹽酸鹽及三碘甲腺原氨酸；及鹽，其包含(但不限於)亞硒酸鈉、磷酸鈉、磷酸鉀、磷酸鈣及/或磷酸鎂。流體培養基可含有上文針對調節表面所描述之抗氧化劑之任何者。可透過一0.22微米過濾器單元(VWR, Cat. No. 73520-986)而無菌過濾流體培養基。在一些實施例中，一適合培養基可包含下列之任何者或可完全由下列之任何者組成：杜爾貝科氏改質伊格爾培養基(ThermoFisher Scientific, Cat # 11960-051)、FreeStyle ^TM培養基(Invitrogen, ThermoFisher Scientific, Cat. No. 11960-051)、RPMI-1640 (GIBCO®, ThermoFisher Scientific, Cat. No. 11875-127)、Hybridoma-SFM (ThermoFisher Scientific, Cat. No. 12045-076)、培養基E (Stem Cell, Cat. No. 3805)、1X CD CHO培養基(ThermoFisher Scientific, Cat. No. 10743-011)、Iscove改質杜爾貝科氏培養基(ThermoFisher Scientific, Cat. No. 12440-061)或CD DG44培養基(ThermoFisher Scientific, Cat. No. 10743-011)。培養基可另外包含胎牛血清(FBS，購自GIBCO®, ThermoFisher Scientific)、熱失活胎牛血清或小牛血清(FCS，Sigma-Aldrich Cat Nos. F2442、F6176、F4135及其他)。FBS可以下列之一濃度存在：約1% v/v至約20% v/v、約1% v/v至約15% v/v、約1% v/v至約10% v/v或約1% v/v至約5% v/v、或該等範圍之任何者內之任何數值。培養基可另外包含人類AB血清(Sigma-Aldrich, Cat. No. S2146)且可以下列之一濃度存在：約1% v/v至約20% v/v、約1% v/v至約15% v/v、約1% v/v至約10% v/v或約1% v/v至約5% v/v、或該等範圍之任何者內之任何數值。培養基可另外包含青黴素-鏈黴素(ThermoFisher Scientific, Cat. No. 15140-163)。筆鏈球菌可以下列範圍內之一濃度存在：約0.01% v/v至約10% v/v、約0.1% v/v至約10% v/v、約0.01% v/v至約5% v/v、約0.1% v/v至約5% v/v、約0.1% v/v至約3% v/v、約0.1% v/v至約2% v/v、約0.1% v/v至約1% v/v或該等範圍之任何者內之任何值。在其他實施例中，培養基可包含遺傳黴素(ThermoFisher Scientific, Cat. No. 101310-035)。遺傳黴素可以下列之一濃度存在：約0.5微克/毫升、約1.0微克/毫升、約5.0微克/毫升、約10.0微克/毫升、約15微克/毫升、約20微克/毫升、約30微克/毫升、約50微克/毫升、約70微克/毫升、約100微克/毫升或此等範圍內之任何值。培養基可包含一緩衝液。該緩衝液可為Good緩衝液之一者。該緩衝液可為(但不限於)(4-(2-羥乙基)-1-哌嗪乙磺酸)(HEPES)(ThermoFisher Scientific, Cat. No. 15630-080)。該緩衝液可以下列之一濃度存在：約1毫莫耳、約3毫莫耳、約5毫莫耳、約7毫莫耳、約9毫莫耳、約10毫莫耳、約12毫莫耳、約15毫莫耳、約20毫莫耳、約40毫莫耳、約60毫莫耳、約100毫莫耳或此等範圍內之任何值。培養基可另外包含穀胺醯胺之二肽取代物，GlutaMAX ^TM(GIBCO® ThermoFisher Scientific, Cat No. 35050-079)。穀胺醯胺之取代物可以下列之一濃度存在：約0.2毫莫耳、約0.5毫莫耳、約0.7毫莫耳、約1.0毫莫耳、約1.2毫莫耳、約1.5毫莫耳、約1.7毫莫耳、約2.0毫莫耳、約2.5毫莫耳、約3.0毫莫耳、約4.0毫莫耳、約7.0毫莫耳或約10.0毫莫耳或此等範圍內之任何值。培養基可包含MEM非必需胺基酸(ThermoFisher Scientific, Cat. No. 10370-088)。MEM非必需胺基酸可以下列之一濃度存在：約0.2毫莫耳、約0.5毫莫耳、約0.7毫莫耳、約1.0毫莫耳、約1.2毫莫耳、約1.5毫莫耳、約1.7毫莫耳、約2.0毫莫耳、約2.5毫莫耳、約3.0毫莫耳、約4.0毫莫耳、約7.0毫莫耳或約10.0毫莫耳或此等範圍內之任何值。培養基可另外含有葡萄糖(ThermoFisher Scientific, Cat. No. 15023-021)。葡萄糖可以下列之一濃度存在：約0.1 g/L、約0.3 g/L、約0.5 g/L、約0.8 g/L、約1.0 g/L、約1.5 g/L、約2.0 g/L、約2.5 g/L、約3.0 g/L、約3.5 g/L、約4.0 g/L、約5.0 g/L、約7.0 g/L、約10.0 g/L或此等範圍內之任何值。培養基可另外包含巰基乙醇(ThermoFisher Scientific, Cat. No. 31350-010)。巰基乙醇可以下列之一濃度存在：約0.001% v/v至約1.5% v/v、約0.005% v/v至約1.0% v/v、約0.01% v/v至約1.0% v/v、約0.15% v/v至約1.0% v/v、約0.2% v/v至約1% v/v或此等範圍內之任何值。培養基可包含OPI培養基添加物，其包含草醯乙酸、丙酮酸及胰島素(Sigma-Aldrich, Cat. No. O-5003)。OPI培養基添加物可以下列之一濃度存在：約0.001% v/v至約1.5% v/v、約0.005% v/v至約1.0% v/v、約0.01% v/v至約1.0% v/v、約0.15% v/v至約1.0% v/v、約0.2% v/v至約1% v/v或此等範圍內之任何值。培養基可含有B-27補充劑(50倍)、無血清培養基(ThermoFisher Scientific, Cat. No. 17504-163)。B-27補充劑可以下列之一濃度存在：約0.01% v/v至約10.5% v/v、約0.05% v/v至約5.0% v/v、約0.1% v/v至約5.0% v/v、約0.5% v/v至約5% v/v或此等範圍內之任何值。如本文中所描述，一培養基或一培養基之一添加物可包含用於產生一調節表面之一或多個Pluronic®聚合物，且可包含Pluronic® L44、L64、P85、F68及F127 (其包含F127NF)。Pluronic®聚合物可以下列之一濃度存在於培養基中：約0.001% v/v至約10% v/v、約0.01% v/v至約5% v/v、約0.01% v/v至約1% v/v或約0.05% v/v至約1% v/v。針對可經提供為一套組之一培養基添加物，濃度可為最終培養基濃度之1倍、5倍、10倍、100倍或約100倍。培養基可包含IL 6 (Sigma-Aldrich, Cat. No. SRP3096-20UG)。IL 6可以下列之一濃度存在：約1 nM、約5 nM、約10 nM、約15 nM、約20 nM、約25 nM、約30 nM、約40 nM、約50 nM或此等範圍內之任何值。培養基可另外包含丙酮酸鈉(ThermoFisher Scientific, Cat. No. 11360-070)。穀胺醯胺之取代物可以下列之一濃度存在：約0.1毫莫耳、約0.02毫莫耳、約0.04毫莫耳、約0.06毫莫耳、約0.08毫莫耳、約0.1毫莫耳、約0.5毫莫耳、約0.7毫莫耳、約1.0毫莫耳、約1.2毫莫耳、約1.5毫莫耳、約1.7毫莫耳、約2.0毫莫耳、約2.5毫莫耳、約3.0毫莫耳、約4.0毫莫耳、約7.0毫莫耳或約10.0毫莫耳或此等範圍內之任何值。 氣體環境系統提供細胞存活所需氣體之一混合物，其包含(但不限於)氧氣及二氧化碳。兩種氣體溶解至流體培養基中，且可由細胞使用，因此隨時間更改一生長室之一隔離區域中之流體培養基之氣體含量。特定言之，二氧化碳含量可隨時間改變，其影響微流體器件中之流體培養基之pH。在一些實驗條件中，可使用非最佳氧氣分壓。 溫度控制在一些實施例中，藉由控制至少一調節表面之溫度而調節(若干)生長室及/或(若干)流動區域之至少一調節表面。系統可包含一組件，其可控制及調變微流體器件之生長室及/或流動區域之至少一調節表面之溫度。系統可包含用於對微流體器件提供溫度調變之帕耳帖加熱、電阻加熱或任何其他適合方法。系統亦可包含感測器及/或回饋組件來將熱輸入控制至一預定範圍。在一些實施例中，至少一調節表面具有至少約25°C、約26°C、約27°C、約28°C、約29°C、約30°C、約31°C、約32°C、約33°C、約34°C、約35°C、約36°C、約37°C、約38°C、約39°C或約40°C之一溫度，且穩定於該溫度處。在一些實施例中，至少一表面具有大於約25°C之一溫度。在其他實施例中，至少一表面具有自約30°C至約40°C、自約35°C至約38°C或自約36°C至約37°C範圍內之一溫度。在一些實施例中，至少一調節表面具有至少約30°C之一溫度。 在培育期間提供灌注之流量控制器流量控制器可在培育週期期間將第一流體培養基灌注於流動區域中(如上文所描述)以將營養素提供至生長室中之細胞且自生長室帶走廢棄物，其中營養素之交換及廢棄物之移除實質上經由擴散而發生。控制器可為與微流體器件分離之一組件或可經併入為微流體器件之部分。流量控制器可經構形以非連續地將培養基灌注於流動區域中。流量控制器可經構形以依一週期性方式或一不規律方式灌注(若干)培養基。在一些其他實施例中，控制器可經構形以約每隔4小時、3小時、2小時、60分鐘、57分鐘、55分鐘、53分鐘、50分鐘、47分鐘、45分鐘、43分鐘、40分鐘、37分鐘、35分鐘、33分鐘、30分鐘、27分鐘、25分鐘、23分鐘、20分鐘、17分鐘、15分鐘、13分鐘、10分鐘、7分鐘或5分鐘將(若干)流體培養基灌注於流動區域中一次。在一些實施例中，控制器可經構形以約每隔5分鐘至約每隔20分鐘灌注流體培養基一次。在其他實施例中，控制器可經構形以約每隔15分鐘至約每隔45分鐘灌注流體培養基一次。在其他實施例中，控制器可經構形以每隔30分鐘至約每隔60分鐘灌注流體培養基一次。在其他實施例中，控制器可經構形以每隔45分鐘至約每隔90分鐘灌注流體培養基一次。在一些其他實施例中，控制器可經構形以每隔60分鐘至約120分鐘灌注流體培養基一次。替代地，控制器可經構形以每隔2小時至每隔6小時灌注流體培養基一次。在一些實施例中，控制器226可經構形以在可為約5秒、約10秒、約15秒、約20秒、約25秒、約30秒、約35秒、約40秒、約45秒、約50秒、約55秒、約60秒、約65秒或約70秒之一時間週期內灌注培養基。在其他實施例中，控制器可經構形以灌注培養基達約1分鐘、約1.2分鐘、約1.4分鐘、約1.5分鐘、約1.6分鐘、約1.8分鐘、約2.0分鐘、約2.2分鐘、約2.4分鐘、約2.5分鐘、約2.6分鐘、約2.8分鐘、約3.0分鐘、約3.2分鐘、約3.4分鐘、約3.5分鐘、約3.6分鐘、約3.8分鐘或約4.0分鐘。在各種實施例中，控制器可經構形以灌注培養基達約5秒至約4分鐘、約10秒至約3.5分鐘、約15秒至約3分鐘、約15秒至約2分鐘、約25秒至約90秒、約30秒至約75秒、約40秒至約2.0分鐘、約60秒至約2.5分鐘、約90秒至約3.0分鐘或1.8分鐘至約4分鐘。流量控制器(圖中未展示)可經構形以依一速率將第一流體培養基灌注於流動區域中，該速率遠大於組分自生長室之隔離區域擴散至流動通道之平均速率。例如，流動區域中之流體流動速率可為約0.009微升/秒、約0.01微升/秒、約0.02微升/秒、約0.03微升/秒、約0.05微升/秒、約0.1微升/秒、約0.2微升/秒、約0.3微升/秒、約0.4微升/秒、約0.5微升/秒、約0.6微升/秒、約0.7微升/秒、約0.8微升/秒、約0.9微升/秒、約1.0微升/秒、約1.1微升/秒、約1.2微升/秒、約1.3微升/秒、約1.4微升/秒、約1.5微升/秒、約1.7微升/秒、約1.9微升/秒、約2.0微升/秒、約2.2微升/秒、約2.4微升/秒、約2.6微升/秒、約2.8微升/秒、約3.0微升/秒、約3.2微升/秒、約3.4微升/秒、約3.6微升/秒、約3.8微升/秒、約4.0微升/秒、約4.2微升/秒、約4.4微升/秒、約4.6微升/秒、約4.8微升/秒、約5.0微升/秒、約6.0微升/秒、約7.0微升/秒、約8.0微升/秒或約9.0微升/秒，其等之任何者係將掃及生長室之一連接區域(但不會掃及(若干)生長室之一隔離區域)之一速率。控制器能夠提供第一流體培養基之一速度，其係流體培養基速度之一非掃及速率，即，低於V _max(微流體器件之最大速度，其避免微流體器件歸因於過度壓力而破裂且限制生長室中之一第二流體培養基與流動區域中之一第一流體培養基之間的組分移動至擴散區)之任何適合速率。在一些實施例中，控制器可經構形以依下列各者透過流動區域而灌注第一流體培養基：約0.05微升/秒、約0.06微升/秒、約0.07微升/秒、約0.08微升/秒、約0.09微升/秒、約0.10微升/秒、約0.11微升/秒、約0.12微升/秒、約0.13微升/秒、約0.14微升/秒、約0.15微升/秒、約0.16微升/秒、約0.17微升/秒、約0.18微升/秒、約0.19微升/秒、約0.20微升/秒、約0.30微升/秒、約0.40微升/秒、約0.50微升/秒、約0.60微升/秒、約0.70微升/秒、約0.80微升/秒、約0.90微升/秒、約1.00微升/秒、約1.10微升/秒、約1.20微升/秒、約1.30微升/秒、約1.40微升/秒、約1.50微升/秒、約1.60微升/秒、約1.70微升/秒、約1.80微升/秒、約1.90微升/秒、約2.00微升/秒、約2.10微升/秒、約2.20微升/秒、約2.30微升/秒、約2.40微升/秒、約2.50微升/秒、約2.60微升/秒、約2.70微升/秒、約2.80微升/秒、約2.90微升/秒或約3.00微升/秒。在一些實施例中，控制器可經構形以依下列各者透過複數個流動區域之各者而灌注第一流體培養基：約0.01微升/秒、約0.02微升/秒、約0.03微升/秒、約0.04微升/秒、約0.05微升/秒、約0.06微升/秒、約0.07微升/秒、約0.08微升/秒、約0.09微升/秒、約0.10微升/秒或約0.11微升/秒。在各種實施例中，流動速率及灌注之持續時間提供流動通道之至少約3個、約4個、約5個、約6個、約7個、約8個、約9個、約10個、約11個、約15個、約20個、約25個、約30個、約35個、約50個、約75個、約100個、約200個、約300個或300個以上體積之第一流體培養基之總量。在各種實施例中，可使用變動持續時間、變動流動速率及灌注停止持續時間來完成灌注，如圖7及圖8之方法中所展示及如下文所討論。 儲液器、培養基調節及引入組 分系統可進一步包含經構形以含有流體培養基之一儲液器，流體培養基可在微流體器件之入口124處引入且可藉由流量控制器而灌注。儲液器可在一上流位置處流體地連接至如本文中所描述之微流體器件之任何者(非限制性實例包含100、200、240、290或400)( 圖 5A 至圖 5E)。可在儲液器中調節流體培養基以含有所要氣體平衡(即，針對一非限制性實例，含有5%二氧化碳之一混合物)，其對受培養之細胞提供最佳化生長，且亦可緩和微流體器件中之pH。在一些實施例中，儲液器可進一步含有不同於微流體器件中之受研究細胞之一細胞群體。此細胞群體可為餵食細胞，其產生微流體器件中之細胞生長及/或存活所需之可溶傳信或生長因子。依此方式，流體培養基可在引入至微流體器件之前針對最佳化生長及/或存活性而調節。使用儲液器來容置餵食細胞群體可防止污染微流體器件中之受培養細胞群體；可將來自餵食細胞之可溶分泌物併入至經傳遞至微流體器件中之流體培養基中，但餵食細胞可不與流體培養基一起向上抽吸。圖5A中展示系統之一儲液器、調節及引入組分之一實施例。在此實施例中，儲液器可為另一微流體器件502，其含有在微流體器件502內調節之流體培養基202 (圖中未展示)。微流體器件502具有一封閉體510及一基底512，其等之至少一者係可透氣的。微流體器件502亦可含有一餵食細胞群體，其經維持使得餵食細胞產生微流體器件500A中之(若干)細胞生長及/或存活所需之可溶生長因子或其他細胞傳信組分。儲液器502可容置於一室516內以提供一5%二氧化碳氣體環境，例如(但不限於)一氣體環境。儲液器502中之流體培養基202吸收穿過儲液器之可透氣壁之氣體混合物(例如空氣中5%二氧化碳)，且亦自餵食細胞吸收可溶分泌物。培養基202藉由泵514而自儲液器502，通過不可透氣連接導管506，經由入口124灌注至微流體器件500A中，且在微流體器件500A之流動通道134中形成流212。在此實施例中，泵連接導管504 (未標記)、轉移連接導管506、基底104或封閉體102均為不可透氣的。流體培養基流212掃及通過微流體器件500A之生長室且容許流體培養基204之廢棄組分自生長室(圖中未展示)擴散出，同時容許組分自流動通道134中之流體培養基202擴散至生長室中。最後，廢流體培養基202' (圖中未展示)經由出口連接導管508中之出口124'而退出微流體器件500A。在另一實施例中，流體培養基202經由泵連接導管504且透過可透氣塊518而轉移至微流體器件500B中，如圖5B中所展示。可透氣塊518併入至封閉體102之上表面中且形成封閉體102之上表面之一部分。由可透氣塊518形成之封閉體102之上表面之部分可位於微流體器件500B之生長室之上流。微流體器件500B容置於一室516內，室516提供經交換至微流體器件500B中之流體培養基中之一氣體環境(例如5%二氧化碳)。另外，室516可對微流體器件500B提供溫度及/或濕度調節。泵連接導管504、封閉體102或基底104均為不可透氣的，且透過可透氣塊518之交換可充當微流體器件500B之「肺」且適當地調節微流體器件500B內之培養基。在此實施例中，流體培養基202可在裝載至泵514中之前於另一組件中經另外調節，且亦可因此含有(例如)來自一餵食細胞培養之分泌物質。在另一實施例中，可透氣塊整合至微流體器件500C之封閉體102之上表面以形成一可透氣區段518'，如圖5C中所展示。流體培養基可如上文針對圖5B之實施例所討論般經調節及經引入且可進一步包含來自一餵食細胞群體之分泌物質。微流體器件500C可容置於含有一氣體環境(例如空氣中5%二氧化碳)之一室516中。氣體環境可橫跨可透氣區段518'交換，可透氣區段518'可為封閉體102之上表面中之一個區段或複數個區段。室516可進一步適當調節器件500C之溫度及濕度。在此實施例中，泵連接導管504、封閉體102 (除可透氣塊518'之外)及基底104可為不可透氣的。在一些實施例中，至少一可透氣區段518'定位於微流體器件500C之一生長室上方。在另一實施例中，至少一可透氣區段518'定位於微流體器件500C之流動區域134上方。在其他實施例中，可透氣區段518'可定位於至少一生長室及至少一流動區域134兩者上方。在一進一步實施例中，可透氣管道504'可用以在將培養基引入至微流體器件500D中之前調節(例如，平衡)流體培養基，如圖5D中所展示。可透氣管道504'之長度可經選擇以提供足以容許一封閉體516內之有效氣體交換及平衡之表面面積，封閉體516可含有諸如(但不限於)空氣中5%二氧化碳之一氣體環境。516之環境可進一步調節可透氣泵連接導管504'內之培養基之溫度及/或濕度。可用於可透氣連接導管之一可透氣材料之一非限制性實例係Teflon® AF。流體培養基可在引入至泵組件514之前藉由與一餵食細胞群體接觸而調節且由此可含有可最佳化微流體器件500D中之(若干)受培養細胞之生長及/或存活性之分泌物質。使用餵食細胞群體之先前調節可發生於室516內或可執行於自身具有對溫度、濕度、pH及/或氣體環境之任何者之環境控制之另一培養組分中。在此實施例中，微流體器件500D之封閉體102及基底104可為不可透氣的。在系統之儲液器、培養基調節及引入組分之又一實施例中，可在能夠放置於一適當氣體環境下之一儲液器502'中調節培養基，如圖5E中所展示。儲液器502'無需為一微流體器件或任何特定類型之培養組件。儲液器502'藉由自一氣體環境源524提供一連接供給526而放置於諸如(例如)空氣中5%二氧化碳之一適當氣體環境下。儲液器502'內之流體培養基具有與自源524提供之氣體環境之氣體交換，且藉此經調節。儲液器502'中之流體培養基亦可含有餵食細胞之一培養基來提供可最佳化微流體器件500E中之受培養細胞之生長及/或存活性之分泌物質。調節流體培養基可自儲液器502'經由轉移連接導管522 (其連接至一泵514上之一閥520)而轉移且可經由連接導管504而由泵514注入至微流體器件500E之通道134中。注入至微流體器件500E中之流體培養基形成流體流212。在通過流動通道134之後，廢流體培養基202'經由出口導管508而退出微流體器件500E。在此實施例中，轉移連接導管522、連接導管504、閥520、泵514、封閉體102及基底104可全部為不可透氣的。在一些實施例中，將源524連接至儲液器502'之連接導管526可實質上為不可透氣的。在其他實施例中，連接導管526無需為實質上不可透氣的，而是可為相對不可透氣的。在圖5E所展示之一些實施例中，氣體(圖中未展示)可為連續流動的或可為脈衝式的，例如，自源524輸入之週期性替換氣體(圖中未展示)可為空氣中5%二氧化碳。在其他實施例中，自源524輸入之氣體可為100%二氧化碳。當使用100%二氧化碳氣體時，少量二氧化碳氣體可注入至儲液器502'之頂部空間(圖中未展示)中以使頂部空間維持一5%二氧化碳混合物。在一些實施例中，當氣體注入至儲液器502'之頂部空間中時，儲液器502'可進一步包含一風扇(圖中未展示)來使注入氣體與已存在於頂部空間(圖中未展示)中之其他氣體組分(圖中未展示)混合。在一些實施例中，當氣體之輸入係脈衝式時，微流體器件500E之蓋102可具有併入或附接於其內之二氧化碳感測器(圖中未展示)。在一些實施例中，100%二氧化碳氣體可自源524輸入以比使用市售空氣中5%二氧化碳氣體混合物節約成本。在其他實施例中，100%二氧化碳氣體可引入至源524中且與空氣混合以製備空氣中之一5%二氧化碳混合物。在上述實施例之任何者中，室516可進一步經加濕使得室之氣體環境不改變微流體器件及/或儲液器中之流體培養基之滲透壓。在另一實施例中，對培養於生長室中之細胞提供適當氣體交換之一替代方法可提供氣體流動通過微流體器件(圖中未展示)之流動區域。適當氣體(例如5%二氧化碳)可直接泵抽或脈動透過流動通道。因為生長室之隔離區域經設計為大多數未被掃及體積，所以定位於隔離區域內之細胞不受移動通過流動通道(掃及區域)之空氣或氣泡干擾。此將提供流動通道中之氣體與生長室內之流體培養基之間的非常快速氣體交換，因為擴散距離相較於(例如)一50 mL錐形管內之擴散距離而言係非常小的。接著，氣體可在任何選定時間量之後由流體培養基替換。氣體流動可依任何所要頻率重複以使溶解氣體組分保持一穩定濃度，該濃度亦對流體培養基之pH產生影響。替代地，非最佳氣體組合物或重複可用以擾動細胞之環境。總言之，存在可用以將調節培養基提供至本文中所描述之微流體器件之生長室中之細胞之各種組件及構形。微流體器件100、200、240、290或400之任何者可與圖5A至圖5E之實施例之任何者一起使用。系統及套組可包含經構形以連接至微流體器件之入口及/或出口之連接導管。連接導管亦可經構形以連接至儲液器及/或泵組件。據此，提供一種用於培養一或多個生物細胞之微流體器件，其包含：一流動區域，其經構形以含有一第一流體培養基之一流動；及至少一生長室，其包括經調節以支援該微流體器件內之細胞生長、存活性、可移植性或其等之任何組合之至少一表面，其中該至少一生長室包含一隔離區域及一連接區域，該隔離區域與該連接區域流體地連接且該連接區域包含至該流動區域之一近端開口。在各種實施例中，該微流體器件之該隔離區域可經構形以含有一第二流體培養基。當該流動區域及該至少一生長室實質上分別填充有該第一流體培養基及該第二流體培養基時，該第二流體培養基之組分可擴散至該第一流體培養基中及/或該第一流體培養基之組分可擴散至該第二流體培養基中；及該第一流體培養基實質上不流動至該隔離區域中。在各種實施例中，該至少一調節表面可經調節以支援該微流體器件內之該一或多個生物細胞之可移植性。在一些實施例中，該調節表面之部分可經構形以支援該微流體器件內之該等生物細胞之可移植性。在一些實施例中，該微流體器件之該至少一調節表面可包含一聚合物，其包含伸烷基醚部分。在其他實施例中，該微流體器件之該至少一調節表面可包含一聚合物，其包括羧酸部分、磺酸部分、核酸部分或膦酸部分。在其他實施例中，該微流體器件之該至少一調節表面可包含一聚合物，其包含糖類部分。在一些實施例中，包含糖類部分之聚合物可為聚葡萄糖。在一些其他實施例中，該微流體器件之該至少一調節表面可包含一聚合物，其包括胺基酸部分。替代地，該微流體器件之該至少一調節表面可包含哺乳動物血清之一或多個組分。哺乳動物血清之該等組分可為一培養基之補充劑。在一些實施例中，該哺乳動物血清可為胎牛血清(FBS)或小牛血清(FCS)。在該微流體器件之各種實施例中，該至少一調節表面可包含糖類部分。在一些實施例中，該至少一調節表面可包含伸烷基醚部分。在其他實施例中，該至少一調節表面可包含胺基酸部分。在一些其他實施例中，該至少一調節表面可包含烷基或全氟烷基部分。在一些實施例中，該等烷基或全氟烷基部分可具有大於10個碳之一主鏈長度。在一些實施例中，該至少一調節表面可包含一部分，其可為：烷基或氟烷基(其包含全氟烷基)部分；單糖或多糖(其可包含(但不限於)聚葡萄糖)；醇類(其包含(但不限於)炔丙醇)；多元醇，其包含(但不限於)聚乙烯醇；伸烷基醚，其包含(但不限於)聚乙二醇；聚電解質(其包含(但不限於)聚丙烯酸或聚乙烯膦酸)；胺基(其包含其衍生物，諸如(但不限於)烷化胺基、羥烷基化胺基、胍鹽及含有一未芳香化氮環原子之雜環基，諸如(但不限於)嗎啉基或哌嗪基)；羧酸，其包含(但不限於)丙炔酸(其可提供羧酸陰離子表面)；膦酸，其包含(但不限於)乙炔基膦酸(其可提供膦酸陰離子表面)；磺酸陰離子；羧基甜菜鹼；磺基甜菜鹼；胺磺酸；或胺基酸。在該微流體器件之各種實施例中，該至少一調節表面可包含共價鍵聯至該微流體器件之一表面之一鍵聯基團，且該鍵聯基團可鍵聯至經構形以支援該微流體器件內之細胞生長、存活性、可移植性或其等之任何組合之一部分。在一些實施例中，該鍵聯基團可為矽烷氧基鍵聯基團。在其他實施例中，該鍵聯基團可為膦酸酯鍵聯基團。在一些實施例中，該鍵聯基團可直接鍵聯至經構形以支援細胞生長、存活性、可移植性或其等之任何組合之該部分。在一些實施例中，該調節表面之該部分可經構形以支援該微流體器件內之該等生物細胞之可移植性。在其他實施例中，該鍵聯基團可直接鍵聯至經構形以支援細胞生長、存活性、可移植性或其等之任何組合之該部分。在其他實施例中，該鍵聯基團可經由一鍵聯子而間接鍵聯至經構形以支援細胞生長、存活性、可移植性或其等之任何組合之該部分。在各種實施例中，該鍵聯子可包含伸三唑基部分。在該微流體器件之各種實施例中，該至少一調節表面可包含兩性離子。在其他實施例中，該至少一調節表面可包含膦酸部分或羧酸部分。在其他實施例中，該調節表面可包含陰離子。在一些其他實施例中，該至少一調節表面可包含胺基或胍部分。在其他實施例中，該至少一調節表面可包含陽離子。在該微流體器件之各種實施例中，該至少一調節表面可包含至少一細胞黏著阻斷分子。該至少一細胞黏著阻斷分子可破壞肌動蛋白纖維形成，阻斷整合素受體，或減弱細胞與DNA污損表面之結合。該至少一細胞黏著阻斷分子可為細胞鬆弛素B、一含RGD之肽或一DNase 1蛋白。在其他實施例中，該至少一細胞黏著阻斷分子可包含多於一種類型之細胞黏著阻斷分子之一組合。在該微流體器件之各種實施例中，該至少一調節表面經構形以被加熱至至少約30°C之一溫度。該至少一調節表面可經構形以穩定於至少約30°C之一溫度處。在該微流體器件之各種實施例中，該微流體器件可進一步包含一微流體通道，其包括該流動區域之至少一部分。在一些實施例中，該至少一生長室之該連接區域可直接通至該微流體通道中。在一些實施例中，該微流體器件之該至少一生長室之該隔離區域可具有足以支援細胞擴增至約100個細胞之一範圍之尺寸。在一些實施例中，可在該至少一生長室中維持不超過1×10 ²個生物細胞。在一些實施例中，該至少一生長室之體積可小於或等於約2×10 ⁶立方微米。在其他實施例中，可在該至少一生長室中維持不超過1×10 ²個生物細胞，且該至少一生長室之體積可小於或等於約1×10 ⁷立方微米。在該微流體器件之各種實施例中，該微流體器件可進一步包含經構形以將該第一流體培養基或該第二流體培養基輸入至該流動區域中之至少一入口及經構形以在該第一培養基自該流動區域退出時接收該第一培養基之至少一出口。在該微流體器件之各種實施例中，該微流體器件可進一步包含具有一介電泳(DEP)構形之一基板，其經構形以將一或多個生物細胞引入至該生長室中或將該一或多個生物細胞移出該生長室。可光學地致動該DEP構形。在該微流體器件之各種實施例中，該微流體器件可進一步包含在該至少一生長室或其之該隔離區域上方之一可變形蓋區域，藉此按壓該可變形蓋區域施加足以使該生物細胞自該隔離區域輸出至該流動區域之一力。在一些實施例中，該微流體器件可包含一蓋，其中該蓋之至少一部分係可透氣的，藉此將一氣體分子源提供至定位於該微流體器件中之一流體培養基。在一些實施例中，該蓋之該可透氣部分可定位於該至少一生長室上方。在一些實施例中，該蓋之該可透氣部分可定位於該流動區域上方。在一些實施例中，該微流體器件可進一步包含在該至少一生長室或其之該隔離區域上方之一可變形蓋區域，藉此按壓該可變形蓋區域施加足以使該生物細胞自該隔離區域輸出至該流動區域之一力。在該微流體器件之各種實施例中，該調節表面可包含一可裂解部分。該可裂解部分可經構形以容許破壞該調節表面，藉此促進該一或多個生物細胞在培養之後之可移植性。在該微流體器件之各種實施例中，該至少一生長室可包含複數個生長室。在該微流體器件之各種實施例中，該一或多個生物細胞可包含複數個生物細胞。在一些實施例中，該一或多個生物細胞可包含一或多個哺乳動物細胞。在一些實施例中，該一或多個生物細胞可包含一或多個融合瘤細胞。在一些實施例中，該一或多個生物細胞可包含一或多個淋巴細胞或白血球細胞。在其他實施例中，該一或多個生物細胞可包含一B細胞、一T細胞、NK細胞、一巨噬細胞或其等之一組合。在各種實施例中，該一或多個生物細胞可包含一或多個貼附細胞。在一些實施例中，該生長室中之該一或多個生物細胞可為一單一細胞且群落可為生物細胞之一選殖群落。 pH 感測器系統可進一步包含連接至微流體器件600 (如圖6中所展示)之至少一入口124及/或至少一出口124'之至少一感測器。替代地，器件600可為器件100、200、240、290、400或500A至500E之任何者。感測器可經構形以在第一流體培養基進入微流體器件600時偵測其之一pH。替代地，感測器可經構形以在第一流體培養基退出微流體器件600時偵測其之一pH。感測器可併入至微流體器件中或其可為能夠附接至微流體器件之一入口124及/或一出口124'或與微流體器件之一入口124及/或一出口124'排成一行之一分離組件。在一些實施例中，pH感測器係一光學感測器。一光學感測器可提供相較於基於電極之台式裝置之一優點，此係因為基於電極之裝置可包含龐大探針以使小量(微升)流體之pH難以或無法量測。類似地，線上流通溶液可歸因於微電極之特性而具有非常長沈降時間(5分鐘至15分鐘)，且可在各使用之前需要外延校準程序。此外，電極會快速劣化以因此需要更多維護。光學感測器可為一整合無電極器件，其包含用於照明之一LED及用於可見色彩偵測之一整合比色感測器。該比色感測器可為一色敏光電晶體。該比色感測器可在可見光波長區域(例如約390奈米至約700奈米)中偵測。經一pH相依染料(諸如(但不限於)酚紅)染色之培養基可提供瞬間且無接觸之光學信號。使用此一光學感測器之一光學無電極量測方法無需與培養基接觸且無需對使用者部分進行校準。光學量測可經校準以消除溫度相依性。另外，一光學感測器之使用最小化使感測器污損之風險以因此減少維護或替換。光源(LED)及色彩感測器之小型化亦使此經得起測試非常小體積液體(＜1微升)及整合至可攜式或手持式儀器中之檢驗。系統可包含藉由LED及光電晶體感測器之控制/監測設備180之驅動電子器件，且可在pH之偵測判定pH在所要範圍外時由控制模組172進一步提供一報警組件。另外，由於色彩偵測之安定時間較快(亞秒)，所以可將此感測器插入於一回饋迴路中以經由調變培養基周圍之一氣體環境中之二氧化碳含量而調節培養基之pH。替代地，控制模組172或控制/監測設備180可進一步提供組件以藉由添加緩衝液及/或酸性或鹼性培養基組分而調變引入流體培養基之pH以將pH校正回所要範圍。在一些實施例中，感測器610連接至流體培養基入口管道606，接近於微流體器件之至少一入口124。管道606可為透明、實質上透明或半透明的。LED 614照射管道606及管道606內之染色流體培養基202a'。整合比色感測器612可監測引入流體培養基之pH，確定pH具有一特定培養實驗之一所要範圍內之一值，且在pH超出該所要範圍時報警。在一些實施例中，感測器610'連接至流體培養基出口管道608，接近於微流體器件之至少一出口124'。管道608可為透明、實質上透明或半透明的。LED 614'照射管道608及管道608內之染色出流流體培養基202a''。整合比色感測器612'可監測引入流體培養基之pH，確定pH具有一特定培養實驗之一所要範圍內之一值，且在pH超出該所要範圍時報警。細胞能夠用於本發明之系統及方法中之一細胞可為任何類型之細胞。例如，細胞可為一胚胎細胞、卵母細胞或精子細胞、幹細胞、先驅細胞或自一組織解離之一細胞、一血細胞、一融合瘤細胞、一培養細胞、來自一細胞系之一細胞、一癌細胞、一受感染細胞、一轉染及/或轉形細胞(細胞系(其包含(但不限於)中國倉鼠卵巢(CHO)細胞)、一報導體細胞或其類似者。細胞可為一哺乳動物細胞或細胞可為非哺乳動物細胞。細胞可包含一細菌、一真菌、一原生動物或受一寄生物種(例如，利什曼原蟲或惡性瘧原蟲)感染之一哺乳動物細胞。在一些實施例中，哺乳動物細胞可為人類、鼠類、豬或所關注之任何其他哺乳動物。在一些實施例中，細胞可來自在培養基中積極生長之一細胞群體或獲得自一新鮮組織樣本(例如，藉由一固體組織樣本之解離，諸如一活體組織檢查或細針抽吸)、血液、唾液、尿液或其他體液。替代地，一或多個生物細胞可來自先前冷凍之其他樣本之一培養基。在一些實施例中，一或多個生物細胞可包含一或多個融合瘤細胞。在其他實施例中，一或多個生物細胞可包含一或多個淋巴細胞或白血球細胞。在一些實施例中，細胞係一B細胞、一T細胞、一NK細胞、一樹突細胞、一巨噬細胞或其他免疫細胞類型或其等之一前驅體，諸如一先驅細胞或一造血幹細胞。在各種實施例中，一或多個生物細胞係一或多個貼附細胞。當將一或多個貼附細胞引入至微流體器件時，額外調節處理可經提供以對貼附細胞提供適當可溶或不可溶環境因子(例如一或多個細胞外間質組分)以容許持久存活及/或細胞增殖。取決於實驗之特定目標，可將僅一個細胞或複數個細胞引入至微流體器件中來培養及/或選殖。當僅一個細胞被引入至系統之一生長室中且根據本文中所描述之方法而培育時，所得擴增群體係最初引入至該生長室中之該細胞之一選殖群落。方法本發明提供一種用於在一系統中培養至少一生物細胞之方法，該系統包含具有至少一生長室及一流動區域之一微流體器件。在亦具有一流動區域之一微流體器件之一生長室中培養一(或若干)細胞可允許營養素、生長因子或其他細胞傳信物種在選定時間週期特定引入以達成細胞生長、存活性或可移植性參數之控制。將該至少一生物細胞引入至具有至少一調節表面之該至少一生長室中，其中該調節表面支援細胞生長、存活性、可移植性或其等之任何組合。在一些實施例中，該調節表面支援該微流體器件內之細胞可移植性。在一些實施例中，可移植性包含防止細胞黏著至該微流體器件。在其他實施例中，可移植性包含對貼附細胞提供將支援細胞生長、存活性、可移植性或其等之任何組合之一調節表面，同時亦允許在一培養週期之後於該微流體器件內移動細胞。該至少一調節表面可為如本文中所描述之任何調節表面。可使用如本文中所描述之數種不同動力來完成該至少一生物細胞之引入，該等動力之部分可容許對將一特定生物細胞放置至該微流體器件上之一特定位置中(例如，放置至一預選定生長室中)進行精確控制。可由本文中所描述之方法完成之細胞放置/移除及營養素/傳信/環境刺激之精確控制難以或無法使用大尺度或其他微流體培養方法來達成。在放置之後，在至少足以使該至少一生物細胞擴增而產生生物細胞之一群落之一長時間週期內培育該至少一生物細胞。當將生物細胞引入至分離生長室中時，所得擴增群落可精確地經識別以進一步用作生物細胞之可分離群組。當將僅一個生物細胞引入至一生長室且允許該生物細胞擴增時，所得群落係生物細胞之一選殖群體。任何適當細胞(其包含(但不限於)如上文所描述之細胞)可用於該等方法中。該微流體器件可為如本文中所描述之微流體器件100、300、400、500A至500E或600之任何者，且該微流體器件可為具有如本文中所描述之組件之任何者之一系統之部分。該至少一生長室可包含複數個生長室，且可使用如本文中所討論之任何適合數目個生長室。在該等方法之一些實施例中，該微流體器件可具有約500個至約1500個生長室、約1000個至約2000個生長室、約1000個至約3500個生長室、約2000個至約5000個生長室、約3000個至約7000個生長室、約5000個至約10000個生長室、約7500個至約15000個生長室、約10000個至約17500個生長室或約12500個至約20000個生長室。在培養一或多個生物細胞之方法中，該至少一調節表面可為如本文中所描述之任何調節表面。該調節表面可共價鍵聯至該微流體器件。在一些實施例中，該調節表面可包含共價鍵聯至該表面之一鍵聯基團，且該鍵聯基團亦可鍵聯至經構形以支援該微流體器件內之該一或多個生物細胞之細胞生長、存活性、可移植性或其等之任何組合之一部分。在一些實施例中，可在輸入該一或多個生物細胞之前提供具有一調節表面之一微流體器件。 引入至少一生物細胞在一些實施例中，將該至少一生物細胞引入至該至少一生長室中可包含：使用具有足以移動該至少一生物細胞之力量之一介電泳(DEP)力。可使用電鑷子(諸如光電鑷子(OET))來產生該DEP力。在一些其他實施例中，將一或多個生物細胞引入至該至少一生長室中可包含：使用流體流動及/或重力(例如，藉由使該微流體器件傾斜，使得該(等)細胞降落至定位於該(等)細胞下方之一生長室中)。在一些實施例中，透過至該微流體器件之一流動區域(例如流動通道)中之一入口124而將該至少一生物細胞引入至該微流體器件中。該流動通道中之培養基流可將該細胞運送至接近於一生長室之一開口之一位置。在將該生物細胞定位於一生長室之一開口接近處之後，可使用本文中所描述之動力(其包含介電泳或重力)之任何者來將該生物細胞移動至該生長室中。介電泳力可包含電致動力或光學致動力，且該等DEP力可進一步由光電鑷子(OET)提供。該至少一生物細胞可移動通過該流動通道而至至少一生長室之一連接區域之近端開口，其中該連接區域直接通至該流動通道/區域且流體地連接至該流動通道/區域。該至少一生長室之該連接區域亦流體地連接至該至少一生長室之一隔離區域。該至少一生物細胞可進一步移動通過該連接區域而至該至少一生長室之該隔離區域中。該至少一生長室之該隔離區域可具有足以支援細胞擴增之尺寸。然而，通常，該生長室之尺寸會將此擴增限制於不超過約1×10 ³個、約5×10 ²個、約4×10 ²個、約3×10 ²個、約2×10 ²個、約1×10 ²個、約50個、約25個、約15個或甚至僅10個培養細胞。在一些實施例中，該隔離區域可具有足以支援細胞擴增至不超過約1×10 ²個、約50個、約25個、約15個或約10個培養細胞之尺寸。吾人已驚奇地發現，可在具有不超過下列各者之一體積之一隔離區域中成功地執行高達約1×10 ²個細胞之細胞培育及/或擴增：約1.0×10 ⁷立方微米、約6×10 ⁶立方微米、約2×10 ⁶立方微米、約1.5×10 ⁶立方微米或約1.0×10 ⁶立方微米。在一些其他實施例中，可在具有不超過約4×10 ⁵立方微米之一體積之一隔離區域中成功地執行高達約1×10 ²個細胞之細胞培育及/或擴增。該生物細胞之大小可取決於細胞類型而自下列各者大幅變動：具有約1微米之一直徑及約1立方微米之一體積之細菌、具有約7微米至約8微米之一直徑及約100立方微米之一體積之一小人類細胞(諸如一血紅細胞)、具有約40微米(非匯合)之一直徑及約2000立方微米之一體積之一永生化細胞系(諸如海拉細胞(HeLa))、具有約25微米至高達約60微米之一直徑及約4700立方微米至約100,000立方微米之一體積之一巨核細胞、或具有約120微米之一直徑及約900,000立方微米之一體積之一人類卵細胞。據此，具有約4×10 ⁵立方微米之一體積之一生長室可容許較大品種之巨核細胞(約1×10 ⁵立方微米之體積)擴增非常少，例如，總共至多不到5個細胞。替代地，相同小生長室(約4×10 ⁵立方微米之體積)可容許細菌細胞(約1立方微米之體積)擴增至約400,000個細菌細胞。方法可進一步包含：將一第一流體培養基引入至該微流體器件之該流動區域之一微流體通道中。在一些實施例中，在引入該至少一生物細胞之前執行引入該第一流體培養基。當在引入該至少一生物細胞之前引入該第一流體培養基時，一流動速率可經選擇使得該第一流體培養基(例如)依任何適合速率自該微流體器件之該流動通道流動至該生長室中。替代地，若該微流體器件已預充有含有過多之一或多個調節試劑之一培養基，則該第一流體培養基依一速率流動至該微流體通道中，使得該第一流體培養基替換該流動區域中之含有(若干)過多調節試劑之任何剩餘培養基。當在將該至少一生物細胞引入至該生長室之後引入該第一流體培養基之流時，該第一流體培養基之流動速率可經選擇以不掃及該隔離區域，其不會使該至少一生物細胞自該隔離區域位移。包圍該至少一生長室之該隔離區域中之該至少一生物細胞的流體培養基係第二流體培養基，其可相同於或不同於該第一流體培養基。在一些實施例中，該第二流體培養基可相同於該第一流體培養基，但在培育步驟期間，細胞廢棄物及耗乏培養基組分可使該第二流體培養基表現為不同於該第一流體培養基。 培育細胞在本文所描述之方法中，在至少足以擴增至少一生物細胞而產生生物細胞之一群落之一長時間週期內培育細胞。該時間週期可經選擇為自約1天至約10天。在其他實施例中，培育週期可延長至超過10天且可延續任何所要週期。由於生長室之隔離區域中之細胞被提供營養素且具有藉由灌注流體培養基而移除之廢棄物，所以細胞可無限期地生長。因為隔離區域填充有擴增細胞群體，所以任何額外擴增將導致擴增生物細胞棲息於生長室之連接區域，連接區域係生長室之一掃及區域。灌注培養基可使擴增生物細胞自生長室之連接區域且隨後自微流體器件掃出。據此，可使存在於生長室之隔離區域中之細胞之數目穩定於取決於生物細胞之大小及生長室之隔離區域之大小的一最大數目處。使一隔離細胞群體中之細胞之最大數目穩定之能力提供相較於細胞培養之其他當前可用方法之一優點，此係因為可消除冗長細胞群體分裂。在一些實施例中，可實施培育達約1天、約2天、約3天、約4天、約5天、約6天、約7天、約8天、約9天、約10天或10天以上。培育週期可在下列範圍內：自約1天至約6天、自約1天至約5天、自約1天至約4天、自約1天至約3天或自約1天至約2天。在其他實施例中，可實施培育達少於約5天、少於約4天、少於約3天或少於約2天。在一些實施例中，可實施培育達少於約3天或少於約2天。在其他實施例中，可實施培育達約3小時、約4小時、約5小時、約6小時、約7小時、約8小時、約9小時、約10小時、約11小時、約12小時、約13小時、約14小時、約15小時、約16小時、約17小時、約18小時、約19小時、約20小時、約21小時、約22小時或約23小時。在培養步驟期間，可在整個培養步驟之一或多個時間點處監測至少一生長室及其內所含之任何細胞之一影像。可將該影像儲存於系統之一處理組件之記憶體中。 灌注細胞在培育步驟期間，存在於生長室之隔離區域內之第二流體培養基可變成耗乏營養素、生長因子或其他生長刺激劑。第二流體培養基可累積細胞廢棄物。另外，因為至少一生物細胞在培育週期期間不斷生長，所以可期望在培育開始時將營養素、生長因子或其他生長刺激劑更改為不同於第一培養基或第二培養基之營養素、生長因子或其他生長刺激劑。如本文中所描述，一微流體器件之一生長室中之培養可給予引入及更改由至少一生物細胞感測之化學梯度(其可更密切接近活體內條件)之特定及選擇性能力。替代地，將由至少一生物細胞感測之化學梯度有意更改為非最佳化條件組可容許細胞在經設計以探索疾病或治療路徑之條件下擴增。因此，方法可包含：在培育步驟期間灌注第一流體培養基，其中經由微流體器件之至少一入口124而引入第一流體培養基且其中經由微流體器件之至少一出口而輸出視情況包括來自第二流體培養基之組分之第一流體培養基。交換第一流體培養基之組分以藉此提供新鮮營養素、可溶生長因子及其類似者及/或交換包圍隔離區域內之(若干)細胞之培養基之廢棄組分發生於實質上在擴散條件下之生長室之掃及區域與未掃及區域之界面處。吾人已驚奇地發現，有效交換產生於實質上無流動條件下。據此，吾人已驚人地發現，成功培育無需連續不斷灌注。因此，灌注可為非連續的。在一些實施例中，灌注係週期性的，且在一些實施例中，灌注係不規律的。灌注週期之間的間斷可具有足以容許隔離區域中之第二流體培養基之組分擴散至流動通道/區域中之第一流體培養基中及/或第一流體培養基之組分擴散至第二流體培養基中之持續時間，第一培養基實質上均不流動至隔離區域中。在另一實施例中，低灌注速率亦可用以獲得生長室之未掃及區域內及生長室之未掃及區域外之流體培養基之組分之有效交換。據此，將至少一生物細胞灌注於一微流體器件之至少一生長室中之一方法展示於圖7中且包含一灌注步驟7002，其中第一流體培養基依一第一灌注速率R ₁透過該微流體器件之一流動區域而流動至流體地連接至該生長室之一流動區域中達一第一灌注時間D ₁。R ₁可經選擇為一非掃及流動速率，如本文中所描述。方法700進一步包含步驟7004：停止流體培養基之流動達一第一灌注停止時間S ₁。使步驟7002及7004重複W次(其中W可為選自1至約1000之一整數)，此後，灌注程序700完成。在一些實施例中，W可為2至約1000之一整數。圖8中展示將至少一生物細胞灌注於一微流體器件之至少一生長室中之另一方法800，其包含一第一灌注循環，該第一灌注循環包含步驟8002：使流體培養基依一第一灌注速率R ₁透過該微流體器件之一流動區域而流動至流體地連接至該生長室之一流動區域中達一第一灌注時間D ₁。R ₁可經選擇為一未掃及流動速率，如本文中所描述。該第一灌注循環包含步驟8004：停止流體培養基之流動達一第一灌注停止時間S ₁。可使該第一灌注循環重複W次，其中W係選自1至約1000之一整數。在該第一灌注循環完成W次重複之後，方法800進一步包含一第二灌注循環，其包含步驟8006：使第一流體培養基依一第二灌注速率R ₂流動達一第二灌注時間D ₂，其中R ₂經選擇為一非掃及流動速率。方法800之該第二灌注循環進一步包含步驟8008：停止流體培養基之流動達一第二灌注停止時間S ₂。此後，方法返回至該第一灌注循環之步驟8002及8004且使組合兩循環灌注程序重複V次，其中V係1至約5000之一整數。W及V之組合可經選擇以滿足所要培育週期終點。在方法700或800之各種實施例中，灌注速率R ₁可為如上文針對流量控制器構形所描述之流體培養基之任何非掃及流動速率。在一些實施例中，R ₁可為約0.009微升/秒、約0.010微升/秒、約0.020微升/秒、約0.030微升/秒、約0.040微升/秒、約0.05微升/秒、約0.06微升/秒、約0.07微升/秒、約0.08微升/秒、約0.09微升/秒、約0.10微升/秒、約0.11微升/秒、約0.12微升/秒、約0.13微升/秒、約0.14微升/秒、約0.15微升/秒、約0.16微升/秒、約0.17微升/秒、約0.18微升/秒、約0.19微升/秒、約0.20微升/秒、約0.30微升/秒、約0.40微升/秒、約0.50微升/秒、約0.60微升/秒、約0.70微升/秒、約0.80微升/秒、約0.90微升/秒、約1.00微升/秒、約1.10微升/秒、約1.20微升/秒、約1.30微升/秒、約1.40微升/秒、約1.50微升/秒、約1.60微升/秒、約1.70微升/秒、約1.80微升/秒、約1.90微升/秒、約2.00微升/秒、約2.10微升/秒、約2.20微升/秒、約2.40微升/秒、約2.50微升/秒、約2.60微升/秒、約2.70微升/秒、約2.80微升/秒、約2.90微升/秒或約3.00微升/秒。在方法800之各種實施例中，第二灌注速率R ₂可為如上文針對流量控制器構形所描述之流體培養基之任何非掃及流動速率。在一些實施例中，R ₂可為0.009微升/秒、0.010微升/秒、0.020微升/秒、0.030微升/秒、0.040微升/秒、0.05微升/秒、0.06微升/秒、0.07微升/秒、0.08微升/秒、0.09微升/秒、0.10微升/秒、0.11微升/秒、0.12微升/秒、0.13微升/秒、0.14微升/秒、0.15微升/秒、0.16微升/秒、0.17微升/秒、0.18微升/秒、0.19微升/秒、0.20微升/秒、0.30微升/秒、0.40微升/秒、0.50微升/秒、0.60微升/秒、0.70微升/秒、0.80微升/秒、0.90微升/秒、1.00微升/秒、1.10微升/秒、1.20微升/秒、1.30微升/秒、1.40微升/秒、1.50微升/秒、1.60微升/秒、1.70微升/秒、1.80微升/秒、1.90微升/秒、2.00微升/秒、2.10微升/秒、2.20微升/秒、2.40微升/秒、2.50微升/秒、2.60微升/秒、2.70微升/秒、2.80微升/秒、2.90微升/秒或3.00微升/秒。可選擇流動速率R ₁及/或R ₂之任何組合。通常，灌注速率R ₂可大於灌注速率R ₁，且可為R ₁之約5倍、約10倍、約20倍、約30倍、約40倍、約50倍、約60倍、約70倍、約80倍、約90倍、約100倍或100倍以上。在一些實施例中，R ₂比R ₁快至少10倍。在其他實施例中，R ₂比R ₁快至少20倍。在又一實施例中，R ₂係R ₁之速率之至少100倍。在方法700或800之各種實施例中，第一灌注時間D ₁可為如上文針對流量控制器構形所描述之任何適合灌注持續時間。在各種實施例中，D ₁可為約5秒、約10秒、約15秒、約20秒、約25秒、約30秒、約35秒、約40秒、約45秒、約50秒、約55秒、約60秒、約65秒、約70秒、約80秒、約90秒、約100秒、約110秒、約120秒、約130秒、約140秒、約150秒、約160秒、約170秒或約180秒。在其他實施例中，D ₁可為一時間範圍，例如約10秒至約40秒，如上文所描述。在一些實施例中，D ₁可為約30秒至約75秒。在其他實施例中，D ₁可為約100秒。在其他實施例中，D ₁可在自約60秒至約150秒之一範圍內。在其他實施例中，D ₁可為約20分鐘、約30分鐘、約40分鐘、約50分鐘、約60分鐘、約80分鐘、約90分鐘、約110分鐘、約120分鐘、約140分鐘、約160分鐘、約180分鐘、約200分鐘、約220分鐘、約240分鐘、約250分鐘、約260分鐘、約270分鐘、約290分鐘或約300分鐘。在一些實施例中，D ₁係約40分鐘至約180分鐘。在方法700或800之各種實施例中，第二灌注時間D ₂可為如上文針對流量控制器構形所描述之任何適合灌注持續時間。在各種實施例中，D ₂可為約5秒、約10秒、約15秒、約20秒、約25秒、約30秒、約35秒、約40秒、約45秒、約50秒、約55秒、約60秒、約65秒、約70秒、約80秒、約90秒或約100秒。在其他實施例中，D ₂可為一時間範圍，例如約5秒至約20秒，如上文所描述。在其他實施例中，D ₂可為約30秒至約70秒。在其他實施例中，D ₂可為約60秒。在方法700或800之各種實施例中，第一灌注時間D ₁可相同於或不同於第二灌注時間D ₂。可選擇D ₁及D ₂之任何組合。在一些實施例中，D ₁及/或D ₂之灌注持續時間可經選擇為短於停止週期S ₁及/或S ₂。在方法700或800之各種實施例中，第一灌注停止時間S ₁可經選擇為如上文針對流量控制器構形之灌注週期之間的一時間間隔所描述之任何適合時間週期。在一些實施例中，S ₁可為約0分鐘、約5分鐘、約10分鐘、約15分鐘、約20分鐘、約25分鐘、約30分鐘、約35分鐘、約40分鐘、約45分鐘、約60分鐘、約65分鐘、約80分鐘、約90分鐘、約100分鐘、約120分鐘、約150分鐘、約180分鐘、約210分鐘、約240分鐘、約270分鐘或約300分鐘。在各種實施例中，S ₁可為如上文針對流量控制器構形之灌注之間的時間間隔所描述之任何適當時間範圍，例如約20分鐘至約60分鐘。在一些實施例中，S ₁可為約10分鐘至約30分鐘。在其他實施例中，S ₁可為約15分鐘。在其他實施例中，S ₁可為約0秒、約5秒、約10秒、約20秒、約30秒、約40秒、約50秒、約60秒、約70秒、約80秒或約90秒。在一些實施例中，S ₁係約0秒。在方法700或800之各種實施例中，第二灌注停止時間S ₂可經選擇為如上文針對流量控制器構形之灌注週期之間的一時間間隔所描述之任何適合時間週期。在一些實施例中，S ₂可為約0分鐘、約5分鐘、約6分鐘、約7分鐘、約8分鐘、約9分鐘、約10分鐘、約20分鐘、約30分鐘、約45分鐘、約50分鐘、約60分鐘、約90分鐘、約120分鐘、約180分鐘、約240分鐘、約270分鐘或約300分鐘。在各種實施例中，S ₂可為如上文針對流量控制器構形之灌注之間的時間間隔所描述之任何適當時間範圍，例如約15分鐘至約45分鐘。在一些實施例中，S ₂可為約10分鐘至約30分鐘。在其他實施例中，S ₂可為約8分鐘或約9分鐘。在其他實施例中，S ₂係約0分鐘。在方法700或800之各種實施例中，第一灌注停止時間S ₁及第二灌注停止時間S ₂可獨立地選自任何適合值。S ₁可相同於或不同於S ₂。在方法800之各種實施例中，重複次數W可經選擇為相同於或不同於重複次數V。在方法700或800之各種實施例中，W可為約1、約4、約5、約6、約8、約10、約12、約15、約18、約20、約24、約30、約36、約40、約45或約50。在一些實施例中，W可經選擇為約1至約20。在一些實施例中，W可為1。在方法800之各種實施例中，V可為約5、約10、約20、約25、約30、約35、約40、約50、約60、約80、約100、約120、約240、約300、約350、約400、約450、約500、約600、約750、約900或約1000。在一些實施例中，V可經選擇為約10至約120。在其他實施例中，V可經選擇為約5至約24。在一些實施例中，V可為約30至約50或可為約400至約500。在方法800之各種實施例中，重複次數W可經選擇為相同於或不同於重複次數V。在方法700或800之各種實施例中，灌注之第一步驟(由步驟7002/7004或8002/8004表示)之一總時間係約1小時至約10小時且W係一整數1。在各種實施例中，灌注之第一步驟之總時間係約9分鐘至約15分鐘。在方法800之各種實施例中，一灌注循環之第二步驟(由步驟8006/8008表示)之一總時間係約1分鐘至約15分鐘或約1分鐘至約20分鐘。在方法700或800之任何者中，灌注方法可持續達生物細胞之整個培育週期，例如約1天、約2天、約3天、約4天、約5天、約6天、約7天、約8天、約9天、約10天或10天以上。在圖8之方法800之另一非限制性實施例中，控制器可經構形以在灌注步驟8002期間將(若干)流體培養基灌注於流動區域中，其具有較長灌注週期D ₁。控制器可依一第一速率灌注流體培養基達下列之一週期：約45分鐘、約60分鐘、約75分鐘、約90分鐘、約105分鐘、約120分鐘、約2.25小時、約2.5小時、約2.45小時、約3.0小時、約3.25小時、約3.5小時、約3.75小時、約4.0小時、約4.25小時、約4.5小時、約4.75小時、約5小時或約6小時。在第一灌注週期D ₁結束時，可停止流體培養基之流動達一停止時間週期S ₁，S ₁可為約0秒、約15秒、約30秒、約45秒、約1分鐘、約1.25分鐘、約1.5分鐘、約2.0分鐘、約3.0分鐘、約4分鐘、約5分鐘或約6分鐘。在一些實施例中，第一流動速率R ₁可經選擇為約0.009微升/秒、約0.01微升/秒、約0.02微升/秒、約0.03微升/秒、約0.05微升/秒、約0.1微升/秒、約0.2微升/秒、約0.3微升/秒、約0.4微升/秒或約0.5微升/秒。可停止流體培養基之流動達少於約1分鐘之一灌注停止週期S ₁或S ₁可為0秒。替代地，S ₁可為約30秒、約1.5分鐘、約2.0分鐘、約2.5分鐘或約3分鐘。接著，可使用一不同灌注速率進行一第二灌注週期D ₂。在一些實施例中，第二灌注速率可高於第一灌注速率。在一些實施例中，第二灌注速率R ₂可選自約1.0微升/秒、約1.1微升/秒、約1.2微升/秒、約1.3微升/秒、約1.4微升/秒、約1.5微升/秒、約1.7微升/秒、約1.9微升/秒、約2.0微升/秒、約2.2微升/秒、約2.4微升/秒、約2.6微升/秒、約2.8微升/秒、約3.0微升/秒、約3.2微升/秒、約3.4微升/秒、約3.6微升/秒、約3.8微升/秒、約4.0微升/秒、約4.2微升/秒、約4.4微升/秒、約4.6微升/秒、約4.8微升/秒、約5.0微升/秒、約6.0微升/秒、約7.0微升/秒、約8.0微升/秒或約9.0微升/秒。第二灌注週期D ₂可為約1秒、約2秒、約3秒、約4秒、約5秒、約6秒、約10秒、約15秒、約30秒、約45秒、約60秒、約65秒、約75秒、約80秒或約90秒。接著可停止灌注達一第二灌注停止週期S ₂，S ₂可為約0秒、約10秒、約20秒、約30秒、約40秒、約50秒、約60秒、約1.5分鐘、約1.75分鐘、約2.0分鐘、約2.5分鐘、約2.75分鐘、約3.0分鐘或約4.0分鐘。在一些實施例中，D ₁可為約2小時、約3小時或約4小時。在各種實施例中，D ₁可為約4小時。在各種實施例中，S ₁可為0秒或少於約1分鐘。第二灌注週期D ₂可為約1秒至約6秒。在一些實施例中，第二灌注停止週期S ₂可為約40秒至約1.5分鐘。據此，提供一種用於將至少一生物細胞灌注於一微流體器件之至少一生長室中之方法，其包含下列步驟：使用一第一灌注步驟來灌注該至少一生物細胞，該第一灌注步驟包含：使一第一流體培養基依一第一灌注速率R ₁流動通過該微流體器件之一流動區域達一第一灌注時間D ₁，其中該流動區域流體地連接至該生長室，其中R ₁經選擇為一非掃及流動速率；停止該第一流體培養基之流動達一第一灌注停止時間S ₁；及使該第一灌注步驟重複W次，其中W係選自1至1000之一整數。該方法可進一步包含下列步驟：使用一第二灌注步驟來灌注該至少一生物細胞，該第二灌注步驟包括：使該第一流體培養基依一第二灌注速率R ₂流動達一第二灌注時間D ₂，其中R ₂經選擇為一非掃及流動速率；停止該第一流體培養基之流動達一第二灌注停止時間S ₂；及使該第一灌注步驟之後該第二灌注步驟重複V次，其中V係1至1000之一整數。第二灌注速率R ₂可大於第一灌注速率R ₁。第一灌注時間D ₁可相同於或不同於第二灌注時間D ₂。第一灌注停止時間S ₁可相同於或不同於第二灌注停止時間S ₂。當執行第二灌注步驟時，重複次數W可相同於或不同於重複次數V。R ₂可比R ₁快至少10倍。替代地，R ₂可比R ₁快至少20倍。R ₂可比R ₁快至少100倍。D ₁可為約30秒至約75秒。在其他實施例中，D ₁可為約40分鐘至約180分鐘或約180分鐘至約300分鐘。在一些其他實施例中，D ₁可為約60秒至約150秒。S ₁可為約10分鐘至約30分鐘。在其他實施例中，S ₁可為約5分鐘至約10分鐘。在其他實施例中，S ₁可為零。在一些實施例中，D ₁可為約40分鐘至約180分鐘，且S ₁可為零。在其他實施例中，D ₁可為約60秒至約150秒，且S ₁可為約5分鐘至約10分鐘。在其他實施例中，D ₁可為約180分鐘至約300分鐘，且S ₁可為零。第一灌注步驟之總時間可為約1小時至約10小時。在其他實施例中，第一灌注步驟之總時間可為約2小時至約4小時。在一些實施例中，W可為大於2之一整數。在一些實施例中，W可為約1至約20。在一些實施例中，D ₂可為約10秒至約25秒。在其他實施例中，D ₂可為約10秒至約90秒。在一些實施例中，S ₂可為約10分鐘至約30分鐘。在其他實施例中，S ₂可為約15分鐘。在一些實施例中，V可為約10至約120。在一些實施例中，V可為約30至約50或可為約400至約500。在一些實施例中，D ₂可為約1秒至約6秒，且S ₂可為0秒。在一些實施例中，D ₂可為約10秒至約90秒且S ₂可為約40秒至約1.5分鐘。在一些實施例中，第二灌注步驟之一重複之一總時間可為約1分鐘至約15分鐘。 調節培養基為提供維持及增強至少一生物細胞之生長及/或存活性之一培養基(例如第一培養基或第二培養基)，第一流體培養基可含有液體組分及氣體組分兩者(例如，氣體組分可溶解於液體組分中)。另外，流體培養基可包含溶解於液體組分中之其他組分，諸如生物分子、維生素及礦物質。任何適合組分可用於流體培養基中，如熟習此項技術者所知。一些非限制性實例係如上文所討論，但可在不背離本文中所描述之方法之情況下使用諸多其他培養基組合物。培養基可或可不含有動物源血清。在一些實施例中，流體培養基可包含一化學成分確定之培養基(至少在接觸細胞或一含細胞之流體之前)，且可進一步為一無蛋白質或無肽之化學成分確定培養基。在一些實施例中，流體培養基可包含一減少血清培養基。在將第一流體培養基引入至微流體器件中之前，可藉由使一初始流體培養基飽含溶解氣體分子而製備第一流體培養基。另外，使初始流體培養基飽含溶解氣體分子可一直持續至將第一流體培養基引入至微流體器件中之時間點。使初始流體培養基飽和可包含：使微流體器件與能夠使初始流體培養基飽含溶解氣體分子之一氣體環境接觸。可使初始流體培養基飽和之氣體分子包含(但不限於)氧氣、二氧化碳及氮氣。第一流體培養基可進一步包含緩和第一流體培養基之一pH。緩和第一流體培養基之pH可(例如)發生於引入溶解氣體分子之前及/或發生於引入溶解氣體分子期間。可藉由添加一緩衝物而實現此緩和。一適合緩衝物之一非限制性實例係HEPES。其他緩衝物可存在於培養基中且可或可不取決於二氧化碳之存在(諸如碳酸緩衝系統)，且可由熟習技術者選擇。細胞生長所需之鹽、蛋白質、碳水化合物、脂類、維生素及其他小分子亦可形成第一流體培養基組合物之部分。在一些實施例中，可在經由入口而引入第一流體培養基之前於一儲液器中執行使第一流體培養基飽含氣體組分。在其他實施例中，可在儲液器與入口之間的一可透氣連接導管中執行使第一流體培養基飽含氣體組分。在其他實施例中，可經由微流體器件之一蓋之一可透氣部分而執行使第一流體培養基飽含氣體組分。在一些實施例中，流體培養基之氣體飽和亦包含：維持氣體交換環境中之濕度，使得微流體器件內之流體培養基之滲透壓在培育期間不改變。第一流體培養基之組合物亦可包含來自一餵食細胞培養基之至少一分泌組分。分泌餵食細胞組分可包含生長因子、內分泌素、細胞激素、小分子、蛋白多糖及其類似者。可在其中執行使第一流體培養基飽含氣體組分之相同儲液器中執行自餵食細胞培養基引入至少一分泌組分，或可在飽和步驟之前完成將至少一分泌組分自餵食細胞培養基引入至第一流體培養基。在一些其他實施例中，第一培養基之組合物亦可包含一(或若干)添加物，其(等)經設計以提供更改流體培養基來測試細胞對該(等)添加物之回應。此(等)添加物可(例如)增強或減弱細胞存活性或生長。在一些實施例中，方法可包含：在經由至少一入口而引入第一流體培養基時偵測第一流體培養基之pH。可在直接接近於入口之一位置處執行偵測pH。在一些實施例中，方法可包含：在經由一出口而輸出第一流體培養基時偵測第一流體培養基之pH。可在直接接近於出口之一位置處執行偵測pH。用於偵測pH之偵測器之任一者或兩者可為一光學感測器。在一些實施例中，若pH偏離一可接受範圍，則偵測器能夠提供一警報。在一些其他實施例中，當由偵測器量測之一pH值偏離一可接受範圍時，可更改第一流體培養基之組合物。在培育步驟期間，可監測至少一生長室及其內所含之任何細胞之一影像。 輸出至少一生物細胞在培育步驟完成之後，可自生長室或其隔離區域輸出至少一生物細胞或細胞群落。輸出可包含：使用足以移動一或多個生物細胞/細胞群落之一強介電泳(DEP)力。可光學地致動或電子地致動該DEP力。例如，微流體器件可包含具有一DEP構形(諸如一光電鑷子(OET)構形)之一基板。在其他實施例中，可使用流體流動及/或重力來使至少一生物細胞或細胞群落自生長室或隔離區域輸出。在其他實施例中，可使用作用於生長室或其隔離區域上方之一可變形蓋區域上之壓縮力來使至少一生物細胞或細胞群落自生長室或隔離區域輸出，藉此致使流體自生長室或隔離區域局部流出。在自生長室或隔離區域輸出至少一生物細胞或細胞群落之後，細胞可自流動區域(例如通道)輸出而離開微流體器件。在一些實施例中，使細胞自流動區域輸出包含：使用足以移動一或多個生物細胞/細胞群落之一強DEP力。可如上文所描述般產生該DEP力。在一些其他實施例中，使細胞自流動區域輸出而離開微流體器件包含：使用流體流動及/或重力來移動細胞。在輸出步驟期間，可監測至少一生長室及其內所含之任何細胞之一影像。 調節至少一表面在一些實施例中，微流體器件具有一調節狀態中之至少一生長室之至少一表面。在其他實施例中，至少一生長室之表面在引入至少一生物細胞之前被調節且可經執行為培養一或多個生物細胞之方法之部分。調節表面可包含：使用一調節試劑(諸如一聚合物)來處理表面。在一些實施例中，提供用於處理一微流體器件(100、300、400、500A至500E及600)之至少一生長室之至少一表面之一方法，其包含下列步驟：使包含過量調節試劑之流體培養基流動至流動通道(圖1A至圖1C、圖2、圖3、圖4A至圖4C)中；培育該微流體器件達一選定時間週期；及替換該通道中之該培養基。在其他實施例中，用於預充一微流體器件之一方法包含下列步驟：使含有一調節試劑之一預充溶液流動至流動通道中；培育該器件達一選定時間週期，藉此調節生長室之至少一表面；及使用一流體培養基替換該通道中之該溶液。該預充溶液可含有如本文中所描述之任何流體培養基。替換調節溶液之流體培養基或具有過量調節試劑之流體培養基可為如本文中所描述之任何培養基且可另外含有細胞。在一些實施例中，可使用包含伸烷基醚部分之一聚合調節試劑來處理至少一表面。具有伸烷基醚部分之該聚合調節試劑可包含任何適合之含伸烷基醚聚合物，其包含(但不限於)上文所討論之含伸烷基醚聚合物之任何者。在一實施例中，可使用兩親性非離子嵌段共聚物來處理生長室之表面，該等兩親性非離子嵌段共聚物包含在聚合物鏈(例如Pluronic®聚合物)內具有不同比率且位於不同位置中之聚環氧乙烷(PEO)及聚環氧丙烷(PPO)亞單元之嵌段。用於產生一調節表面之特定Pluronic®聚合物包含Pluronic® L44、L64、P85、F68及F127 (其包含F127NF)。在其他實施例中，可使用包含羧基部分之一聚合調節試劑來處理表面。適合之含羧酸聚合調節試劑之非限制性實例係如上文所討論且任何適當之含羧酸聚合調節試劑可用以處理表面。在其他實施例中，可使用包含糖類部分之一聚合調節試劑來處理表面。適合之含糖類聚合調節試劑之非限制性實例係如上文所討論且任何適當之含糖類聚合調節試劑可用以處理表面。在其他實施例中，可使用包含磺酸部分之一聚合調節試劑來處理表面。適合之含磺酸聚合調節試劑之非限制性實例係如上文所討論且任何適當之含磺酸聚合調節試劑可用以處理表面。在其他實施例中，可使用包含胺基酸部分之一聚合調節試劑來處理表面。適合之含胺基酸聚合調節試劑之非限制性實例係如上文所討論且任何適當之含胺基酸聚合調節試劑可用以處理表面。含胺基酸聚合調節試劑可包含一蛋白質。在一些實施例中，使用一蛋白質來處理表面，其中該蛋白質可包含發現於一哺乳動物血清中之一組分或包含一哺乳動物血清之部分。在其他實施例中，使用一哺乳動物血清之組分來處理表面。在一些實施例中，可使用一細胞培養基補充劑(諸如B-27®補充劑((50倍)，來自ThermoFisher Scientific, Cat# 17504044之無血清培養基))來處理表面。該哺乳動物血清可為胎牛血清(FBS)。替代地，該哺乳動物血清可為小牛血清(FCS)。在其他實施例中，可使用包含核酸部分之一聚合調節試劑來處理表面。適合之含核酸聚合調節試劑之非限制性實例係如上文所討論且任何適當之含核酸聚合調節試劑可用以處理表面。在一些實施例中，一種以上聚合調節試劑之一混合物可用以處理生長室之表面。在一些其他實施例中，調節步驟可包含：使用至少一細胞黏著阻斷分子來處理至少一生長室之至少一表面。在一些實施例中，可在自微流體器件輸出細胞之前執行使用至少一細胞黏著阻斷分子來處理至少一生長室之至少一表面之步驟。在一些實施例中，調節步驟可包含：使用至少一細胞黏著阻斷分子來預培育細胞。在一些實施例中，至少一細胞黏著阻斷分子可用以破壞肌動蛋白纖維形成。在一些實施例中，細胞黏著阻斷分子可為細胞鬆弛素B。在其他實施例中，至少一細胞黏著阻斷分子可阻斷整合素受體。在一些實施例中，細胞黏著阻斷分子可包含肽，其含有一RGD基元。在一些其他實施例中，至少一細胞黏著阻斷分子可減弱細胞與DNA污損表面之結合。可減弱細胞與DNA污損表面之結合之細胞黏著阻斷分子可包含一DNase 1蛋白。在其他實施例中，至少一細胞黏著阻斷分子可包含一小分子纖連蛋白抑制劑。在其他實施例中，至少一細胞黏著阻斷分子可為一抗體，例如一抗B1整合素抗體。在一些實施例中，至少一細胞黏著阻斷分子可包含多於一種類型之細胞黏著阻斷分子之一組合。在其他實施例中，調節包含：將生長室之表面加熱至約30°C之一溫度。在一些實施例中，方法包含：將表面加熱至至少約25°C、約26°C、約27°C、約28°C、約29°C、約30°C、約31°C、約32°C、約33°C、約34°C、約35°C、約36°C、約37°C、約38°C、約39°C或約40°C之一溫度。在一些實施例中，方法包含：將表面加熱至大於約25°C之一溫度。在其他實施例中，方法包含：將表面加熱至自約30°C至約40°C、自約35°C至約40°C或自約36°C至約38°C之範圍內之一溫度。在一些實施例中，方法包含：將表面加熱至至少約30°C之一溫度。在一些實施例中，加熱表面包含藉由使用一聚合物來處理表面而調節之至少一表面。 選殖群 體本文中所描述之方法亦包含其中將僅一個生物細胞引入至至少一生長室之方法。提供用於在包含一微流體器件之一系統中選殖一生物細胞之一方法，該微流體器件具有：一流動區域，其經構形以含有一第一流體培養基之一流動；及至少一生長室，其包含一隔離區域及一連接區域，該隔離區域與該連接區域流體地連接且該連接區域包含至該流動區域之一近端開口，該方法包含下列步驟：將該生物細胞引入至該至少一生長室中，其中該至少一生長室經構形以具有經調節以支援細胞生長、存活性、可移植性或其等之任何組合之至少一表面；及在至少足以擴增該生物細胞而產生生物細胞之一選殖群體之一長時間週期內培育該生物細胞。在一些實施例中，該系統可為如本文中所描述之任何系統。該微流體器件可為如本文中所描述之任何微流體器件。在用於選殖一生物細胞之該方法之一些實施例中，該至少一調節表面可包含共價鍵聯至該表面之一鍵聯基團，且該鍵聯基團可鍵聯至經構形以支援該微流體器件內之該一或多個生物細胞之細胞生長、存活性或可移植性之一部分。在一些實施例中，該鍵聯基團可包含矽烷氧基鍵聯基團。在其他實施例中，該鍵聯基團可包含膦酸酯鍵聯基團。在一些實施例中，該鍵聯基團可間接鍵聯至經構形以支援細胞生長、存活性、可移植性或其等之任何組合之該部分。在其他實施例中，該鍵聯基團可直接鍵聯至經構形以支援細胞生長、存活性、可移植性或其等之任何組合之該部分。該鍵聯基團可經由連接至一鍵聯子而間接鍵聯至經構形以支援細胞生長、存活性或可移動性之該部分。在一些實施例中，該鍵聯基團可經由連接至一鍵聯子之一第一端而間接鍵聯至經構形以支援細胞生長、存活性或可移動性之該部分。在一些實施例中，該鍵聯子可進一步包含一直鏈部分，其中該直鏈部分之一主鏈包括選自矽原子、碳原子、氮原子、氧原子、硫原子及磷原子之任何組合之1個至200個非氫原子。在一些實施例中，該直鏈部分之該主鏈可包含一或多個伸芳基部分。在其他實施例中，該鍵聯子可包含伸三唑基部分。在一些實施例中，該伸三唑基部分可中斷該鍵聯子之該直鏈部分或可連接至該鍵聯子之該直鏈部分之一第二端處。在各種實施例中，經構形以支援細胞生長及/或存活性及/或可移植性之該部分可包含：烷基或氟烷基(其包含全氟烷基)部分；單糖或多糖(其可包含(但不限於)聚葡萄糖)；醇類(其包含(但不限於)炔丙醇)；多元醇，其包含(但不限於)聚乙烯醇；伸烷基醚，其包含(但不限於)聚乙二醇；聚電解質(其包含(但不限於)聚丙烯酸或聚乙烯膦酸)；胺基(其包含其衍生物，諸如(但不限於)烷化胺基、羥烷基化胺基、胍鹽及含有一未芳香化氮環原子之雜環基，諸如(但不限於)嗎啉基或哌嗪基)；羧酸，其包含(但不限於)丙炔酸(其可提供羧酸陰離子表面)；膦酸，其包含(但不限於)乙炔基膦酸(其可提供膦酸陰離子表面)；磺酸陰離子；羧基甜菜鹼；磺基甜菜鹼；胺磺酸；或胺基酸。在一些實施例中，該至少一調節表面包括烷基或全氟烷基部分。在其他實施例中，該至少一調節表面包括伸烷基醚部分或聚葡萄糖部分。在各種實施例中，該方法可進一步包含下列步驟：調節該至少一生長室之該至少一表面。在一些實施例中，調節包含：使用支援該微流體器件內之細胞可移植性之一或多個試劑來處理該至少一表面。在一些實施例中，該調節可包含：使用包含一聚合物之一調節試劑來處理該至少一生長室之該至少一表面。在一些實施例中，該聚合物可包含伸烷基醚部分。在一些實施例中，該聚合物可包含羧酸部分。在一些實施例中，該聚合物可包含糖類部分。在其他實施例中，該聚合物可包含磺酸部分。在其他實施例中，該聚合物可包含胺基酸部分。在進一步實施例中，該聚合物可包含核酸部分。在一些實施例中，該調節可包含：使用哺乳動物血清之一或多個組分來處理該至少一生長室之該至少一表面。在一些實施例中，該哺乳動物血清可為胎牛血清(FBS)或小牛血清(FCS)。在各種實施例中，調節可包含：使用至少一細胞黏著阻斷分子來處理該至少一生長室之至少一表面。在一些實施例中，該至少一細胞黏著阻斷分子可包含一含RGD之肽。在其他實施例中，該至少一細胞黏著阻斷分子可為細胞鬆弛素B、一整合素抗體、纖連蛋白之一抑制劑或一DNase 1蛋白。在各種實施例中，調節可包含：使用多於一種類型之細胞黏著阻斷分子之一組合來處理該至少一生長室之至少一表面。在各種實施例中，該調節可包含：將該至少一生長室之該至少一表面加熱至約30°C之一溫度。在各種實施例中，該方法可進一步包含下列步驟：將一第一流體培養基引入至該微流體器件之該流動區域之一微流體通道中。在一些實施例中，可在引入該生物細胞之前執行引入該第一流體培養基。在一些實施例中，將該生物細胞引入至該至少一生長室中可包含：使用具有足以移動該生物細胞之力量之一介電泳(DEP)力。在一些實施例中，可光學地啟動該DEP力。在一些實施例中，可由光電鑷子(OET)產生該DEP力。在一些其他實施例中，將該生物細胞引入至該至少一生長室中可包含：使用流體流動及/或重力。在一些實施例中，將該生物細胞引入至該至少一生長室中可進一步包含：將該生物細胞引入至該至少一生長室之一隔離區域中。在一些實施例中，該至少一生長室之該隔離區域可具有足以支援細胞擴增至不超過1×10 ²個細胞之尺寸。在一些實施例中，該隔離區域可至少實質上填充有一第二流體培養基。在一些實施例中，該流動區域可流體地連接至該至少一生長室之一連接區域之一近端開口，且其中該連接區域亦可進一步流體地連接至該生長室之該隔離區域。在各種實施例中，該方法可進一步包含下列步驟：在該培育步驟期間灌注該第一流體培養基，其中可經由該微流體器件之至少一入口而引入該第一流體培養基且其中可經由該微流體器件之至少一出口而輸出視情況包括來自該第二流體培養基之組分之該第一流體培養基。在一些實施例中，灌注可為非連續的。在一些其他實施例中，灌注可為週期性的。在其他實施例中，灌注可為不規律的。在一些實施例中，可依足以容許該隔離區域中之該第二流體培養基之組分擴散至該流動區域中之該第一流體培養基中及/或該第一流體培養基之組分擴散至該隔離區域中之該第二流體培養基中之一速率執行灌注該第一流體培養基；及該第一流體培養基可實質上不流動至該隔離區域中。在一些實施例中，可約每隔10分鐘至約每隔30分鐘執行灌注該第一流體培養基達約45秒至約90秒之一持續時間。在一些實施例中，可執行灌注該第一流體培養基達約2小時至約4小時之一持續時間。在一些實施例中，培育該至少一生物細胞之該時間週期可為自約1天至約10天。在一些實施例中，該第一流體培養基之一組合物可包含液體組分及氣體組分。在各種實施例中，該方法可進一步包含下列步驟：在將該第一流體培養基引入至該微流體器件中之前使該第一流體培養基飽含溶解氣體分子。在各種實施例中，該方法可進一步包含下列步驟：使該微流體器件與能夠使該第一流體培養基或該第二流體培養基飽含溶解氣體分子之一氣體環境接觸。在各種實施例中，該方法可進一步包含下列步驟：在引入溶解氣體分子之後緩和該第一流體培養基之一pH。在一些實施例中，可在經由該入口而引入該第一流體培養基之前於一儲液器中執行使該第一流體培養基飽含該等氣體組分，在該儲液器與該入口之間的一可透氣連接器中執行使該第一流體培養基飽含該等氣體組分，或經由該微流體器件之一蓋之一可透氣部分而執行使該第一流體培養基飽含該等氣體組分。在一些實施例中，該第一流體培養基之一組合物可包含來自一餵食細胞培養基之至少一分泌組分。在各種實施例中，該方法可進一步包含下列步驟：在經由該至少一出口而輸出該第一流體培養基時偵測該第一流體培養基之pH。在一些實施例中，可在直接接近於該至少一出口之一位置處執行該偵測步驟。在各種實施例中，該方法可進一步包含下列步驟：在經由該至少一入口而引入該第一流體培養基時偵測該第一流體培養基之pH。在一些實施例中，感測器可為一光學感測器。在各種實施例中，該方法可進一步包含下列步驟：更改該第一流體培養基之一組合物。在各種實施例中，該方法可進一步包含下列步驟：監測該至少一生長室及其內所含之任何細胞之一影像。在各種實施例中，該生物細胞可為一哺乳動物細胞。在一些實施例中，該生物細胞可為一免疫細胞。在一些實施例中，該生物細胞可為一淋巴細胞或一白血球。在一些實施例中，該生物細胞可為一B細胞、一T細胞、一NK細胞、巨噬細胞或樹突細胞。在一些實施例中，該生物細胞可為一貼附細胞。在一些實施例中，該生物細胞可為一融合瘤細胞。在一些實施例中，該生物細胞可為複數個生物細胞且該至少一生長室係複數個生長室。在各種實施例中，該方法可進一步包含下列步驟：將該複數個生物細胞之僅一者移動至該複數個生長室之各者中。在選殖一生物細胞之該方法之一些實施例中，該調節表面可進一步包含一可裂解部分。該方法可包含下列步驟：在使該選殖群體之一或多個生物細胞自該生長室或其之該隔離區域輸出之前使該可裂解部分裂解。在各種實施例中，該方法可進一步包含下列步驟：使該選殖群體之一或多個生物細胞自該生長室或其之該隔離區域輸出。在一些實施例中，輸出可包含：使用足以移動該一或多個生物細胞之一強介電泳(DEP)力。在一些實施例中，光學地致動該DEP力。在一些實施例中，可由光電鑷子(OET)產生該DEP力。在一些實施例中，輸出可包含：使用流體流動及/或重力。在一些實施例中，輸出可包含：使用作用於該生長室或其之該隔離區域上方之一可變形蓋區域上之壓縮力。在各種實施例中，該方法可進一步包含下列步驟：使該選殖群體之一或多個生物細胞自該流動區域輸出而離開該微流體器件。在一些實施例中，輸出可包含：使用足以移動該一或多個生物細胞之一強DEP力。在一些實施例中，光學地致動該DEP力。在一些實施例中，可由光電鑷子(OET)產生該DEP力。在一些實施例中，輸出可包含：使用流體流動及/或重力。套組本發明可提供用於培養一生物細胞之套組，其中該套組包含：一微流體器件，其具有經構形以含有一第一流體培養基之一流動之一流動區域及至少一生長室；及一表面調節試劑。在此實施例中，未預處理該至少一生長室來調節該至少一生長室之至少一表面，而是藉由在引入(若干)細胞之前使用該表面調節試劑來處理而產生該調節表面。本發明亦提供用於培養一生物細胞之其他套組，其中該套組包含一微流體器件，其具有：一流動區域，其經構形以含有一第一流體培養基之一流動；及至少一生長室，其包括一隔離區域及一連接區域，其中該隔離區域與該連接區域流體地連接且該連接區域包括至該流動區域之一近端開口；及其中該至少一生長室進一步包括經調節以支援細胞生長、存活性、可移植性或其等之任何組合之至少一表面。本發明亦提供用於培養一生物細胞之其他套組，其包含一微流體器件，該微流體器件包含：一流動區域，其經構形以含有一第一流體培養基之一流動；及至少一生長室，其包含一隔離區域及一連接區域，其中該隔離區域與該連接區域流體地連接且該連接區域具有至該流動區域之一近端開口；其中該至少一生長室具有至少一表面，其具有一表面改質配位基。替代地，本發明可提供用於培養一生物細胞之套組，其中該套組包含：一微流體器件，其具有一流動區域(其經構形以含有一第一流體培養基之一流動)及至少一生長室(其具有可支援細胞生長、存活性、可移植性或其等之任何組合之至少一調節表面)；及一表面調節試劑。該等套組之任何者之微流體器件可為微流體器件100、200、240、290、400、500A至500E或600之任何者且具有上文所描述之特徵之任何者。該等套組之任何者之微流體器件可進一步包含一微流體通道，其包含該流動區域之至少一部分；且該器件可進一步包含一生長室，其具有直接通至該微流體通道中之一連接區域。該生長室可進一步包含一隔離區域。該隔離區域可流體地連接至該連接區域且可經構形以含有一第二流體培養基，其中當該流動區域及該至少一生長室實質上分別填充有一第一流體培養基及一第二流體培養基時，該第二流體培養基之組分擴散至該第一流體培養基中及/或該第一流體培養基之組分擴散至該第二流體培養基中；及該第一培養基實質上不流動至該隔離區域中。在該等套組之任何者之各種實施例中，生長室可如同圖1A至圖1C、圖2、圖3及圖4A至圖4C之生長室124、126、128、130、244、246、248或436般構形，其中該生長室之該隔離區域可具有經構形以支援不超過約1×10 ³個、約5×10 ²個、約4×10 ²個、約3×10 ²個、約2×10 ²個、約1×10 ²個、約50個、約25個、約15個或約10個培養細胞之一體積。在其他實施例中，該生長室之該隔離區域具有可支援高達約10個、約50個或約1×10 ²個細胞之一體積。如上文所討論，該等生長室之任何構形可用於該等套組之該等微流體器件之該等生長室中。在該等套組之任何者之各種實施例中，該等生長室之大小可經構形以維持不超過1×10 ²個生物細胞，其中該等生長室之體積可不超過1×10 ⁷立方微米。在其他實施例中，可維持不超過1×10 ²個生物細胞，該等生長室之體積可不超過5×10 ⁶立方微米。在其他實施例中，可維持不超過50個生物細胞，且該等生長室之體積可不超過1×10 ⁶立方微米或不超過5×10 ⁵立方微米。在該等套組中，該等微流體器件可具有任何數目個如上文所討論之生長室。該等套組之任何者之微流體器件可進一步包含經構形以將流體培養基(例如第一流體培養基或第二流體培養基)輸入至該流動區域中之至少一入口及經構形以在該流體培養基(例如廢第一流體培養基)自該流動區域退出時接收該流體培養基之至少一出口。該等套組之任何者之微流體器件可進一步包含具有複數個DEP電極之一基板，其中該基板之一表面形成該生長室及該流動區域之一表面。該複數個DEP電極可經構形以產生足以使一或多個生物細胞(例如一選殖群體)移動至一生長室或其之該隔離區域中或使一生物細胞培養基之一或多個細胞移出一生長室或其之該隔離區域之一強介電泳(DEP)力。可光學地致動該等DEP電極且因此光學地致動該DEP力。此等光學致動之DEP電極可為虛擬電極(例如歸因於入射光而具有增強導電性之一非晶矽基板之區域)、光電晶體或由一對應光電晶體接通或切斷之電極。替代地，可電致動該DEP電極且因此電致動該DEP力。在一些其他實施例中，該微流體器件可進一步包含具有複數個電晶體之一基板，其中該基板之一表面形成該生長室及該流動區域之一表面。該複數個電晶體能夠產生足以將該生物細胞引入至該生長室或其之該隔離區域中或將一生物細胞培養基之一或多個細胞移出該生長室或其之該隔離區域之一強介電泳(DEP)力。可光學地致動該複數個電晶體之各者，且可由光電鑷子產生該DEP力。該等套組之任何者之微流體器件可進一步包含在該至少一生長室或其隔離區域上方之一可變形蓋區域，藉此按壓該可變形蓋區域施加使一或多個生物細胞(例如一選殖群體)自該生長區域輸出至該流動區域之一力。該等套組之任何者之微流體器件可經構形以具有實質上不可透氣之一蓋。替代地，該蓋之一部分之全部可經構形為可透氣的。該蓋之該可透氣部分可滲透二氧化碳、氧氣及氮氣之至少一者。在一些實施例中，該蓋(或其之一部分)可滲透二氧化碳、氧氣或氮氣之一者以上之一組合。該等套組之任何者可進一步包含經構形以含有一流體培養基之一儲液器。該儲液器可流體地連接至本文中所描述之微流體器件之任何者。該儲液器可經構形使得存在於該儲液器中之該流體培養基可由能夠使該流體培養基飽含溶解氣體分子之一氣體環境接觸。該儲液器可進一步經構形以含有與該流體培養基流體接觸之一餵食細胞群體。該等套組之任何者可包含經構形以連接至該微流體器件之一入口及/或一出口之至少一連接導管。該連接導管亦可經構形以連接至一儲液器或一流量控制器，諸如一泵組件。該連接導管可為可透氣的。該可透氣連接導管可滲透二氧化碳、氧氣及氮氣之至少一者。在一些實施例中，該可透氣導管可滲透二氧化碳、氧氣或氮氣之一者以上之一組合。該等套組之任何者可進一步包含經構形以偵測一第一流體培養基之一pH之一感測器。該感測器可連接至該微流體器件之一入口或其之一附接連接導管。替代地，該感測器可整合至該微流體器件。該感測器可連接至流體培養基進入該微流體器件之點之接近處。該套組可包含經構形以偵測該微流體器件之該輸出處之流體培養基之一pH之一感測器。該感測器可連接至該微流體器件之一出口或其之一附接連接導管。替代地，該感測器可整合至該微流體器件。該感測器可連接至流體培養基退出該微流體器件之點之接近處。無論該感測器是否附接至該微流體器件之該入口及/或該出口，其均可為一光學感測器。一光學感測器可包含一LED及一整合比色感測器，該整合比色感測器可視情況為一色敏光電晶體。該套組可進一步包含驅動電子組件來控制該pH感測器且自該pH感測器接收輸出。該套組可進一步包含一pH偵測劑。該pH偵測劑可為可在可見光下偵測到之一pH敏感染料。該等套組之任何者亦可包含一培養基，其具有能夠增強該微流體器件上之生物細胞存活性之組分。此等組分可為此項技術中已知之任何適合培養基組分，其包含上文針對流體培養基組分所討論之組分之任何者。該等套組之任何者可進一步包含用以偵測一生物細胞或一細胞群體之一狀態之至少一試劑。經構形以偵測細胞之狀態之試劑在此項技術中係熟知的，且可用以(例如)偵測一細胞是否活著或死亡，是否分泌所關注之一物質(諸如抗體、細胞激素或生長因子)，或是否具有所關注之細胞表面標記。此等試劑可用於(但不限於)本文中所描述之套組及方法中。針對本文中所提供之套組之任何者，該等套組之組分可位於分離容器中。針對提供於溶液中之套組之組分之任何者，該等組分可以約為用於本發明之方法中之濃度之1倍、5倍、10倍、100倍或1000倍之一濃度存在。針對其中未預處理微流體器件之至少一生長室來調節至少一生長室之至少一表面之套組，藉由使用表面調節試劑來處理而產生調節表面；或針對包含一微流體器件(其具有：一流動區域，其經構形以含有一第一流體培養基之一流動；及至少一生長室，其具有可支援細胞生長、存活性、可移植性或其等之任何組合之至少一調節表面)及一表面調節試劑之套組，可使用一表面調節試劑來預調節該生長室之該表面。該表面調節試劑可包含一聚合物，其可為用作一表面調節試劑之上文所描述之聚合物之任何一或多者。在一些實施例中，該表面調節試劑可包含一聚合物，其具有伸烷基醚部分、羧酸部分、磺酸部分、胺基酸部分、核酸部分、糖類部分或其等之任何組合。該表面調節試劑可包含一PEO-PPO嵌段共聚物，諸如Pluronic®聚合物(例如L44、L64、P85或F127)。在一些實施例中，該表面調節試劑可包含哺乳動物血清之一或多個組分。該哺乳動物血清可為胎牛血清(FBS)或小牛血清(FCS)。替代地，用以調節該生長室之該表面之該表面調節試劑可包含於該套組中，與該微流體器件分離。在該套組之其他實施例中，包含一預調節微流體器件及不同於用以調節該生長室之該表面之該表面調節試劑的一表面調節試劑。該不同表面調節試劑可為上文所討論之表面調節試劑之任何者。在一些實施例中，一種以上表面調節試劑包含於該套組中。在具有一微流體器件之該等套組(其中未預處理該微流體器件之該至少一生長室來調節該至少一表面)之各種實施例中，該套組亦可包含適合用於培養該一或多個生物細胞之一培養基。在一些實施例中，該套組亦可包含一培養基添加物，其包括能夠加強該生長室之一表面之調節之一試劑。該培養基添加物可包含如上文所討論之一調節試劑或增強該至少一生長室之該至少一表面支援細胞生長、存活性、可移植性或其等之任何組合之能力之另一化學物種。此可包含生長因子、內分泌素、抗氧化劑或維生素及其類似者。該套組亦可包含經構形以灌注至少該第一流體培養基之一流量控制器，其可為該微流體器件之一分離組件或可經併入為該微流體器件之部分。該控制器可經構形以非連續地灌注該流體培養基。因此，該控制器可經構形以依一週期性方式或依一不規律方式灌注該流體培養基。在另一態樣中，提供一種用於培養一生物細胞之套組，其包含一微流體器件，其具有：一流動區域，其經構形以含有一第一流體培養基之一流動；及至少一生長室，其包括一隔離區域及一連接區域，其中該隔離區域與該連接區域流體地連接且該連接區域包括至該流動區域之一近端開口；且其中該至少一生長室進一步包括經調節以支援細胞生長、存活性、可移植性或其等之任何組合之至少一表面。該微流體器件可為如本文中所描述之任何微流體器件，且可具有如本文中所描述之生長室之任何者。該微流體器件可具有一基板，其具有本文中所描述之任何種類之一DEP構形。可光學地致動該DEP構形。該微流體器件之該基板可具有包含式1或式2之基板組合物(如本文中所描述)之一表面，且具有如上文所描述之全部特徵。

式1 式2 該套組之該微流體器件之該至少一調節表面可包含糖類部分、伸烷基醚部分、胺基酸部分、烷基部分、氟烷基部分(其可包含全氟烷基部分)、陰離子部分、陽離子部分及/或兩性離子部分。在一些實施例中，該微流體器件之該調節表面可包含糖類部分、伸烷基醚部分、烷基部分、氟烷基部分或胺基酸部分。該等烷基或全氟烷基部分可具有大於10個碳之一主鏈長度。在一些實施例中，用以支援細胞生長、存活性、可移植性或其等之任何組合之該調節表面可包含：烷基或氟烷基(其包含全氟烷基)部分；單糖或多糖(其可包含(但不限於)聚葡萄糖)；醇類(其包含(但不限於)炔丙醇)；多元醇，其包含(但不限於)聚乙烯醇；伸烷基醚，其包含(但不限於)聚乙二醇；聚電解質(其包含(但不限於)聚丙烯酸或聚乙烯膦酸)；胺基(其包含其衍生物，諸如(但不限於)烷化胺基、羥烷基化胺基、胍鹽及含有一未芳香化氮環原子之雜環基，諸如(但不限於)嗎啉基或哌嗪基)；羧酸，其包含(但不限於)丙炔酸(其可提供羧酸陰離子表面)；膦酸，其包含(但不限於)乙炔基膦酸(其可提供膦酸陰離子表面)；磺酸陰離子；羧基甜菜鹼；磺基甜菜鹼；胺磺酸；或胺基酸。在該套組之一些實施例中，該調節表面可包含共價鍵聯至該微流體器件之一表面之一鍵聯基團，且該鍵聯基團可鍵聯至經構形以支援該微流體器件內之該一或多個生物細胞之細胞生長、存活性、可移植性或其等之任何組合之該部分。該鍵聯基團可為矽烷氧基鍵聯基團。替代地，該鍵聯基團可為膦酸酯鍵聯基團。在該套組之一些實施例中，該調節表面之該鍵聯基團可直接鍵聯至經構形以支援細胞生長、存活性、可移植性或其等之任何組合之該部分。在其他實施例中，該鍵聯基團可經由一鍵聯子而間接鍵聯至經構形以支援細胞生長、存活性、可移植性或其等之任何組合之該部分。該鍵聯基團可經由連接至一鍵聯子之一第一端而間接鍵聯至經構形以支援細胞生長、存活性、可移植性或其等之任何組合之該部分。該鍵聯子可進一步包含一直鏈部分，其中該直鏈部分之一主鏈包括選自矽原子、碳原子、氮原子、氧原子、硫原子及磷原子之任何組合之1個至200個非氫原子。在該套組之一些實施例中，該調節表面之該鍵聯子可進一步包含伸三唑基部分。該可裂解部分經構形以容許破壞該調節表面，藉此促進該生物分子之可移植性。該套組可進一步包含經構形以使該調節表面之該可裂解部分裂解之一試劑。在該套組之各種實施例中，該套組可進一步包含一表面調節試劑。在一些實施例中，該表面調節試劑可包含一聚合物，其包括下列之至少一者：伸烷基醚部分、羧酸部分、磺酸部分、膦酸部分、胺基酸部分、核酸部分或糖類部分。在一些其他實施例中，該表面調節試劑包括一聚合物，其包括伸下列之至少一者：烷基醚部分、胺基酸部分或糖類部分。在一些其他實施例中，該調節表面可包含一可裂解部分。在該套組之其他實施例中，該表面調節試劑包括至少一細胞黏著阻斷分子。在一些實施例中，該至少一細胞黏著阻斷分子可破壞肌動蛋白纖維形成，阻斷整合素受體，或減弱細胞與DNA污損表面之結合。在一些實施例中，該至少一細胞黏著阻斷分子可為細胞鬆弛素B、一含RGD之肽、一DNase 1蛋白、一纖連蛋白抑制劑或一整合素抗體。在一些實施例中，該至少一細胞黏著阻斷分子可包含多於一種類型之細胞黏著阻斷分子之一組合。在該套組之各種實施例中，該表面調節試劑可包含哺乳動物血清之一或多個組分。該哺乳動物血清可為胎牛血清(FBS)或小牛血清(FCS)。在該套組之各種實施例中，該套組可進一步包含適合用於培養該一或多個生物細胞之一培養基。在一些實施例中，該套組可包含一培養基添加物，其包含經構形以加強生長室之該至少一表面之調節之一試劑。該培養基添加物可包含如上文所討論之一調節試劑或增強該至少一生長室之該至少一表面支援細胞生長、存活性、可移植性或其等之任何組合之能力之另一化學物種。此可包含生長因子、內分泌素、抗氧化劑或維生素及其類似者。在該套組之各種實施例中，該套組可包含用以偵測該一或多個生物細胞之一狀態之至少一試劑。在又一態樣中，提供一種用於培養一生物細胞之套組，其包含用於培養一或多個生物細胞之一微流體器件，該微流體器件包含：一流動區域，其經構形以含有一第一流體培養基之一流動；及至少一生長室，其包含一隔離區域及一連接區域，其中該隔離區域與該連接區域流體地連接且該連接區域具有至該流動區域之一近端開口；且該至少一生長室具有至少一表面，其具有一表面改質配位基。該微流體器件可為如本文中所描述之任何微流體器件。該表面可包含具有一介電泳(DEP)構形之一基板。該DEP構形可為本文中所描述之任何DEP構形。可光學地致動該DEP構形。該基板係具有如本文中所描述之一表面改質配位基之任何基板，且可具有式3之一結構，且可包含如上文所描述之全部特徵：

式3 在具有一微流體器件(其具有包含一表面改質配位基之至少一表面)之該套組之各種實施例中，該表面改質配位基可共價鍵聯至該基板之該表面之氧化物部分。該表面改質配位基可包含一反應部分。該表面改質配位基之該反應部分可為疊氮基、胺基、溴基、硫醇基、活性酯基、琥珀醯亞胺基或炔基部分。該表面改質配位基可經由一鍵聯基團而共價鍵聯至該等氧化物部分。在一些實施例中，該鍵聯基團可為矽烷氧基部分。在其他實施例中，該鍵聯基團可為磷酸酯部分。該鍵聯基團可經由一鍵聯子而間接連接至該表面改質配位基之該反應部分。該鍵聯子可包含一直鏈部分，其中該直鏈部分之一主鏈包含選自矽原子、碳原子、氮原子、氧原子、硫原子及磷原子之任何組合之1個至100個非氫原子。在一些實施例中，該表面改質配位基可包含一或多個可裂解部分。該一或多個可裂解部分可經構形以容許在形成一微流體器件之一調節表面之後破壞該調節表面，藉此促進該一或多個生物細胞在培養之後之可移植性。在具有一微流體器件(其具有包含一表面改質配位基之至少一表面)之該套組之一些實施例中，該套組可進一步包含一調節改質劑，其包含經構形以支援細胞生長、存活性、可移植性或其等之任何組合之一第一部分及經構形以與該表面改質配位基之反應部分反應之一第二部分，該調節改質劑可具有式5之一結構且具有如本文中所描述之特徵之任何者：

式5 該第二部分可經構形以在與該套組之該微流體器件之該表面改質配位基之該反應部分反應之後將該表面改質配位基轉換成經構形以支援生長室內之一或多個生物細胞之細胞生長、存活性、可移植性或其等之任何組合之一調節表面。該第一部分可包含伸烷基氧化物部分、糖類部分、烷基部分、全氟烷基部分、胺基酸部分、陰離子部分、陽離子部分或兩性離子部分。在一些實施例中，該第一部分可包含：烷基或氟烷基(其包含全氟烷基)部分；單糖或多糖(其可包含(但不限於)聚葡萄糖)；醇類(包含(但不限於)炔丙醇)；多元醇，其包含(但不限於)聚乙烯醇；伸烷基醚，其包含(但不限於)聚乙二醇；聚電解質(其包含(但不限於)聚丙烯酸或聚乙烯膦酸)；胺基(其包含其衍生物，諸如(但不限於)烷化胺基、羥烷基化胺基部分、胍鹽及含有一未芳香化氮環原子之雜環基，諸如(但不限於)嗎啉基或哌嗪基)；羧酸，其包含(但不限於)丙炔酸(其可提供羧酸陰離子表面)；膦酸，其包含(但不限於)乙炔基膦酸(其可提供膦酸陰離子表面)；磺酸陰離子；羧基甜菜鹼；磺基甜菜鹼；胺磺酸；或胺基酸。該第二部分可為胺基、羧酸、炔基、疊氮基、醛基、溴基或硫醇基部分。在一些實施例中，該調節改質劑之該第一部分或一鍵聯子L' (如上文針對式5所描述)可包含一可裂解部分。該可裂解部分可經構形以容許破壞該調節表面，藉此促進該生物細胞之可移植性。在一些實施例中，該套組可進一步包含經構形以使該調節表面之該可裂解部分裂解之一試劑。在具有一微流體器件(其具有包含一表面改質配位基之至少一表面)之該套組之一些實施例中，該套組可進一步包含一表面調節試劑。在具有一微流體器件(其具有包含一表面改質配位基之至少一表面)之該套組之一些實施例中，該表面調節試劑可包含一聚合物，其包括下列之至少一者：伸烷基醚部分、羧酸部分、磺酸部分、膦酸部分、胺基酸部分、核酸部分或糖類部分。在一些其他實施例中，該表面調節試劑包括一聚合物，其包括下列之至少一者：伸烷基醚部分、胺基酸部分或糖類部分。在一些其他實施例中，該調節表面可包含一可裂解部分。在具有一微流體器件(其具有包含一表面改質配位基之至少一表面)之該套組之一些實施例中，該表面調節試劑包括至少一細胞黏著阻斷分子。在一些實施例中，該至少一細胞黏著阻斷分子可破壞肌動蛋白纖維形成，阻斷整合素受體，或減弱細胞與DNA污損表面之結合。在一些實施例中，該至少一細胞黏著阻斷分子可為細胞鬆弛素B、一含RGD之肽、一DNase 1蛋白、一纖連蛋白抑制劑或一整合素抗體。在一些實施例中，該至少一細胞黏著阻斷分子可包含多於一種類型之細胞黏著阻斷分子之一組合。在具有一微流體器件(其具有包含一表面改質配位基之至少一表面)之該套組之一些實施例中，該表面調節試劑可包含哺乳動物血清之一或多個組分。該哺乳動物血清可為胎牛血清(FBS)或小牛血清(FCS)。在具有一微流體器件(其具有包含一表面改質配位基之至少一表面)之該套組之一些實施例中，該套組可進一步包含適合用於培養該一或多個生物細胞之一培養基。在一些實施例中，該套組可進一步包含一培養基添加物，其包含經構形以加強生長室之該至少一表面之調節之一試劑。該培養基添加物可包含如上文所討論之一連接劑或增強該至少一生長室之該至少一表面支援細胞生長、存活性、可移植性或其等之任何組合之能力之另一化學物種。此可包含生長因子、內分泌素、抗氧化劑或維生素及其類似者。在具有一微流體器件(其具有包含一表面改質配位基之至少一表面)之該套組之一些實施例中，該套組可進一步包含用以偵測該一或多個生物細胞之一狀態之至少一試劑。實例實例 1. 一 K562 紅白血 病 細胞之培養及生長 材料：K562細胞(一人類永生化骨髓性白血病細胞系)獲得自美國菌種保存中心(ATCC)(catalog ATCC® CCl-243 ^TM)且經提供為一懸浮細胞系。藉由播種1×10 ³個活細胞/mL且在37°C處使用5%二氧化碳氣體環境來培育而維持培養。使細胞分裂成1×10 ⁶個細胞/mL或使細胞每隔2天至3天分裂。在5%二甲基亞碸(DMSO)/95%完全生長培養基中冷凍細胞。 培養基：Iscove之改質杜爾貝科氏培養基(ATCC® Catalog No. 30-2005)+10%胎牛血清(Hyclone, Cat# SH30071.2)經組合以產生完全生長培養基。當在培育週期期間灌注時，在將完全生長培養基引入至微流體器件中之前使用空氣中之5%二氧化碳來連續調節完全生長培養基。 預充溶液：含有0.1% Pluronic® F127 (Life Technologies® Cat# P6866)之完全生長培養基 系統及微流體器件：由Berkeley Lights公司製造。系統包含至少一流量控制器、溫度控制器、流體培養基調節及泵抽組件、用於光啟動DEP構形之光源、微流體器件、安裝台及一攝影機。用於此實驗中之微流體器件之生長室具有約1.4×10 ⁵立方微米之一體積。流動通道之橫截面積係約4×10 ³平方微米。微流體器件具有8個通道。培養準備：將微流體器件裝載至系統上且使用具有15 psi之100%二氧化碳沖洗微流體器件達5分鐘。在二氧化碳沖洗之後，依5微升/秒即時灌注預充溶液通過微流體器件達8分鐘。接著，使完全生長培養基依5微升/秒流動通過微流體器件達5分鐘。 培養條件：使微流體器件之溫度維持於37°C處。在培養實驗之整個週期中依0.001微升/秒之一恆定速率灌注培養基。使用重力來使一單一K562細胞裝載至微流體器件之一生長室中。展示裝載細胞之後之t=0小時處之生長室之一照片(參閱圖10A)。箭頭1002指向生長室中之單一細胞之位置。在16小時培養完成之後，將細胞擴增至2個細胞之一群體，如該時間點所拍攝之一照片中所展示(參閱圖10B)。箭頭1004指向生長室中之兩個細胞之位置。在34小時培養完成之後，將細胞群體增加至總共四個細胞，如圖10C之照片中所展示。箭頭1006及1008指向定位於生長室內之兩個細胞之兩個群組之各者。在54小時培養完成之後，將K562細胞群體增加至總共八個細胞，如圖10D之照片中所展示。箭頭1010及1012指向定位於生長室內之細胞群組之任一側處之細胞。在70小時培養完成之後，將K562細胞群體增加至總共16個細胞，如圖10E之照片中所展示。箭頭1014、1016及1018指向該群組之細胞。將K562之一選殖擴增群體提供於微流體器件之生長室中。實例 2.一 OKT3 融合瘤細胞之培養及生長 材料：OKT3細胞(一鼠科骨髓性融合瘤細胞系)獲得自ATCC (ATCC® Cat. # CRL-8001™)。細胞經提供為一懸浮細胞系。藉由播種約1×10 ⁵個活細胞/mL至約2×10 ⁵個活細胞/mL且在37°C處使用空氣中之5%二氧化碳(作為氣體環境)來培育而維持培養。使細胞每隔2天至3天分裂。計數OKT3細胞數目及存活性且將細胞密度調整至5×10 ⁵/ml以裝載至微流體器件。培養基：500 mL Iscove之改質杜爾貝科氏培養基(ATCC® Catalog No. 30-2005)、200 mL胎牛血清(ATCC® Cat. #30-2020)及1 mL青黴素-鏈黴素(Life Technologies® Cat. # 15140-122)經組合以產生培養基。透過一0.22μm過濾器而過濾完全培養基且在使用之前將完全培養基儲存於4°C之避光處。當在培育週期期間灌注時，在將培養基引入至微流體器件中之前使用空氣中之5%二氧化碳連續調節培養基。預充 溶液：含有0.1% Pluronic® F127 (Life Technologies® Cat# P6866)之培養基。 系統及微流體器件：由Berkeley Lights公司製造。系統包含至少一流量控制器、溫度控制器、流體培養基調節及泵抽組件、用於光啟動DEP構形之光源及投射器、微流體器件、安裝台及一攝影機。用於此實驗中之微流體器件之生長室具有約1.5×10 ⁶立方微米之一體積。一流動通道之橫截面積係8×10 ³平方微米，且總共六個通道存在於微流體器件上。 培養準備：將微流體器件裝載至系統上且使用具有15 psi之100%二氧化碳來沖洗微流體器件達5分鐘。在二氧化碳沖洗之後，依8微升/秒即時灌注預充溶液通過微流體器件，直至灌注2.5 mL之一總體積通過微流體器件。接著，使培養基依8微升/秒流動通過微流體器件，直至灌注總共1 mL之培養基通過微流體器件。圖11A之照片中展示引入細胞之前預備微流體器件。一列之四個生長室沿照片之底部延伸。 培養條件：使微流體器件之溫度維持於37°C處。在培養實驗之整個週期中使用一可變灌注方法來灌注培養基，該可變灌注方法包含依0.01微升/秒之一初始4小時灌注週期，接著為依8微升/秒之一短時高速灌注(約3秒)，接著為約小於1分鐘之一短灌注停止週期。在整個培養實驗中持續進行包含交替灌注速率及一停止之此循環。藉由重力而將一單一OKT3細胞引入至生長室中。圖11B中展示時間t=0處之具有一個細胞之生長室之一照片，其中箭頭1102指向左邊第二室，且尤其指向室內之單一細胞，其中駐留細胞之區域進一步由圓圈圈出。圖12A至圖12C展示培養實驗中之隨後時間點處之微流體器件之照片且演示形成一選殖群體之細胞擴增。圖12A之照片拍攝於一天培養完成時且箭頭1202指向左邊第二室(單一OKT3細胞之引入點)中之一群組之約4個細胞。圖12B係在2天培養完成之後拍攝之一照片且箭頭1204指向左邊第二室中之一進一步增殖細胞群體。圖12C係在3天培養完成之後拍攝之一照片，且箭頭1206展示由培養單一OKT3細胞引起之眾多擴增OKT3細胞。圖13A至圖13C展示在三天培養完成之後(即，在圖12C之時間點之後)之微流體器件之照片，且演示使用由光電鑷子產生之一介電泳力來輸出擴增OKT3細胞之一選集。圖13A中將引發介電泳力之光圖案(即，箭頭1302所指向之一光阱)展示為圍繞細胞之一白框。由光學致動之介電泳力使細胞自生長室之底部朝向流動通道移動。圖13B之照片展示擴增OKT3細胞進一步移動至流動區域中。仍將細胞陷留於光阱中且迫使細胞與光阱一起移動(箭頭1304)。圖13C之照片展示在將擴增細胞完全移動至流動區域中之後釋放擴增細胞(箭頭1306)。此等細胞藉由使用光學致動之DEP力、重力或流體流動而自微流體器件輸出此等細胞用於進一步研究或擴增。此實驗演示藉由使用本文中所描述之器件及方法而提供之選擇性、精確度及靈活性。實例 3. 使用一無血清培養基來移除貼附細胞以調節一微流體器件之表面 系統及微流體器件：如同實例1，使用具有約7×10 ⁵立方微米之一體積之生長室。 預充方案：使250微升之100%二氧化碳依12微升/秒之一速率流入。此後，使含有0.1% Pluronic® F27 (Life Technologies® Cat# P6866)之250微升之PBS依12微升/秒流入。預充之最後步驟包含：使250微升之PBS依12微升/秒流入。接著，引入培養基。 灌注方案：灌注方法係下列兩種方法之任一者： 1. 依0.01微升/秒灌注2小時；依2微升/秒灌注64秒；且重複。 2. 依0.02微升/秒灌注100秒；停止流動500秒；依2微升/秒灌注64秒；且重複。 培養基 ：無血清培養基(ThermoFisher Scientific, Cat. No. 12045-096)。 系統及微流體器件 ：在培養之後自一微流體器件之流動通道移除貼附細胞之能力演示為：在36°C處於具有一調節培養基添加物(B-27®補充劑)(2% v/v)之一無血清培養基中預培育貼附細胞(其可為(例如)購自AddexBio, Cat. No. C000605之JIMT1細胞)達30分鐘。在預培育之後，將貼附細胞引入至流動通道，停止流動，且培育貼附細胞達2小時至約24小時之一週期。在化驗結束之後，依5微升/秒之一速率引入無血清培養基之流動。約750微升之流量(其表示約150倍微流體器件體積)通過微流體器件，全部貼附JIMT1細胞自流動通道輸出而離開微流體器件。此實驗展示：可含有補充組分(諸如市售B27)之無血清培養基可防止併入貼附報導體細胞之一化驗之過程期間之黏著，且容許貼附細胞自微流體器件輸出。實例 4. 使用一調節混合物來移除貼附細胞以調節一微流體器件之表面 貼附細胞：如同上述實例3。 培養基：具有添加組分(其包含(但不限於) FBS (購自ThermoFisher Scientific, Cat. No. 16000-036)及青黴素-鏈黴素(ThermoFisherScientific Cat. No. 15140-163))之一無血清培養基(ThermoFisher Scientific, Cat. No. 12045-076)。 調節混合 物：細胞鬆弛素B (Sigma Aldrich, Catalog No. C2743-200UL)、DNaseI (New England Biosciences Cat No. M0303S)及RGD三肽(Santa Cruz Biotechnology Cat No. sc-201176)。 貼附細胞製備：使用調節混合物來改質培養基以具有下列之一最終濃度：4微莫耳細胞鬆弛素B、0.1單位/微莫耳DNaseI及1毫莫耳RGD三肽。在將貼附細胞輸入至微流體器件之前於36°C處培育貼附細胞達30分鐘。 系統及微流體器件。如同上文，使用具有約7×10 ⁵立方微米之一體積之生長室。在培養之後自一微流體器件之流動通道移除貼附細胞(例如JIMT1細胞)之能力演示為：預培育使用一調節混合物來預培育之貼附細胞群體。值得注意地，調節混合物之使用容許將含血清培養基(諸如用於此實例中之培養基)用於微流體環境內，同時仍給予貼附細胞之移除。將預培育貼附細胞引入至微流體器件之流動通道中，且培育貼附細胞達2小時至約24小時之一週期。在化驗結束之後，依5微升/秒之一速率引入培養基之流動。約750微升之流量(其表示約150倍微流體器件體積)通過微流體器件，全部貼附細胞自流動通道輸出而離開微流體器件。此實驗展示：調節混合物可防止黏著且容許輸出貼附細胞。實例 5. 具有調節表面之微流體器件之製備 針對全部製備：微流體器件：如同上述實例1，由Berkeley Lights公司製造，且被直接使用。無論何種情況，具有圖案化聚矽氧(PPS)之矽基板及ITO/玻璃基板係在合成調節表面之前於一Nordson Asymtek電漿 清洗器中清洗(100 W電力、50 s)之氧氣電漿。 A. 全氟烷基矽烷氧 基 調節表面 材料：十七氟-1,1,2,2-四氫十二烷基三甲氧基矽烷獲得自Gelest (Cat. No. SIH5841.5)且被直接使用。直接使用MgSO ₄·7H ₂O (Acros)。 製備之方法：藉由使經組裝之微流體器件在降低壓力下於升高溫度處暴露於十七氟-1,1,2,2-四氫十二烷基三甲氧基矽烷及水蒸汽而化學地改質經組裝之微流體器件。添加300微升十七氟-1,1,2,2-四氫十二烷基三甲氧基矽烷及0.5克MgSO ₄·7H ₂O (水源)以分離在一清潔乾燥6''玻璃真空乾燥器之底部中之鋁舟。將微流體器件支撐於矽烷試劑及水合鹽(水源)上方之一多孔板上。在室溫處將乾燥器泵抽至750 mTorr且密封乾燥器。接著，將乾燥劑放置至一110°C爐中達24小時。接著，具有全氟烷基調節表面之微流體器件自乾燥器移除且被使用。在一些實驗中，在將微流體器件安裝至印刷電路板之前化學地改質微流體器件。 B. 聚葡萄糖調節表面 材料：藉由使用疊氮化鈉來置換溴化物部分而由11-溴十一烷基三甲氧基矽烷(Gelest)合成11-疊氮十一烷基三甲氧基矽烷。在一典型反應中，將4.00克11-溴十一烷基三甲氧基矽烷(Gelest)添加至含有2.00克疊氮化鈉(Sigma-Aldrich)之一60微升無水二甲基甲醯胺(DMF)(Acros)溶液中。在氮氣下於室溫處攪拌溶液達24小時。接著，過濾溶液，且使用乾燥戊烷(Acros)來提取濾液。粗製11-疊氮十一烷基三甲氧基矽烷產物藉由旋轉蒸發而濃縮且藉由兩次連續真空蒸餾而純化。二苯并環辛炔(DBCO)改質聚葡萄糖(MW：約3000 Da)購自Nanocs且被直接使用。 準備方法 ： 引入一表面改質配位基。藉由使經組裝微流體器件之表面在降低壓力下於升高溫度處暴露於11-疊氮十一烷基三甲氧基矽烷及水蒸汽而化學地改質經組裝微流體器件之表面。添加300微升11-疊氮十一烷基三甲氧基矽烷及0.5克MgSO ₄·7H ₂O(水源)以分離一清潔乾燥6''玻璃真空乾燥器之底部中之鋁舟。將微流體器件支撐於矽烷及水合鹽(水源)上方之一多孔板上。在室溫處將乾燥器泵抽至750 mTorr且密封乾燥器。接著，將乾燥器放置至一110°C爐中達24小時。接著，自乾燥器移除具有表面改質配位基(11-疊氮十一烷基矽烷氧基部分)之微流體晶片。在一些實驗中，在將微流體器件安裝至印刷電路板之前化地學改質微流體器件。 聚葡萄糖調節表面之引入：藉由在氣相沈積之後使含有166微莫耳DBCO聚葡萄糖之一至少250微升水溶液流動通過具有表面改質疊氮配位基之微流體器件而使疊氮基封端之微流體器件表面與DBCO聚葡萄糖反應。允許反應在室溫處進行達至少1小時。接著，藉由使至少250微升DI水流動通過晶片而沖洗晶片。 C. 聚乙二醇 (PEG) 調節表面 材料：如上文般合成11-疊氮十一烷基三甲氧基矽烷。炔基改質PEG (MW：約5000 Da)購自JenKem且被直接使用。抗壞血酸鈉及五水合硫酸銅購自Sigma-Aldrich且被直接使用(THPTA (三(3-羥丙基三唑基甲基)胺)銅催化點擊試劑(Glen Research))。 準備之方法 ： 引入一表面改質配位基。如上文般製備具有11-疊氮十一烷基矽烷氧基表面改質配位基之微流體晶片。 PEG 調節表面之引入：藉由使含有333微莫耳炔基改質PEG、500微莫耳硫酸銅、500微莫耳THPTA配位基及5毫莫耳抗壞血酸鈉之一至少250微升水溶液流動通過具有11-疊氮十一烷基矽烷氧基表面改質配位基之微流體器件而使微流體器件之疊氮基封端之表面與炔基改質PEG反應。允許反應在室溫處進行達至少1小時。接著，藉由使至少250微升去離子水流動通過器件而沖洗具有一PEG調節表面之微流體器件。 D. 具有至表面之膦酸酯鍵聯基團之烷基改質表面 材料：膦酸十八酯購自Sigma Aldrich且被直接使用。丙酮及乙醇購自Sigma Aldrich。 製備之方法：使微流體器件之表面在35°C處暴露於一10毫莫耳之膦酸十八酯無水乙醇溶液中達48小時。在沈積之後使用乙醇及DI水來充分沖洗具有經由膦酸酯鍵聯基團而附接之烷基調節表面之所得微流體器件。實例 6. 一調節微流體表面上之 T 淋巴細胞之培養及輸出 材料：CD3+細胞，其來自AllCells公司且依1珠粒/1細胞之一速率與抗CD3/抗CD28磁珠(Dynabeads®, Thermofisher Scientific, Cat. No. 11453D)混合。在37°C處於一5% CO ₂培育箱中之於相同於培養實驗本身之培養基中培育混合物達5小時。在培育之後，使T細胞/珠粒混合物再懸浮而供使用。 培養基：RPMI-1640 (GIBCO®，ThermoFisher Scientific, Cat. No. 11875-127)、10% FBS、2%人類AB血清(50 U/ml IL2；R&D系統) 預充 程序：如同上述實例3。 灌注方案：如同上述實例3。 系統及微流體器件：如同上述實例3。生長室具有約7×10 ⁵立方微米之一體積。 調節表面：微流體器件具有如上文所描述般製備之一共價鍵聯聚葡萄糖調節表面。藉由使再懸浮液流動通過一流體入口且進入微流體通道而將(T細胞+珠粒)懸浮液引入至微流體器件中。藉由使晶片傾斜且允許重將動T細胞/珠粒拉至生長室中而停止流動且使T細胞/珠粒隨機裝載至生長室中。在將T細胞/珠粒裝載至生長室之後，灌注培養基通過奈流體晶片之微流體通道達4天之一時間週期。圖14A展示微流體器件之生長室之聚葡萄糖調節表面上之T細胞之生長。相對於一類似微流體器件(資料未展示)之一非調節表面而改良聚葡萄糖調節表面上之T細胞之生長。接著，藉由重力(例如，使微流體器件傾斜)而使T細胞自生長室移除。圖14B展示在一20分鐘週期結束時自生長室移除之程度(其演示使擴增T細胞輸出至流動通道中之卓越能力)，其相較於自一類似微流體器件之一非調節表面移除T細胞之程度而改良。接著，使T細胞自微流體器件(圖中未展示)移除。本文中所展示之實例係例示性的且絕不限制[實施方式]中所描述之方法及裝置之範疇。 This application is a non-provisional application claiming rights under 35 U.S.C. 119(e) to U.S. Provisional Application No. 62/151,325, filed April 22, 2015, the entire contents of which are incorporated herein by reference case. Microfluidic environments provide the opportunity to provide a cell or group of cells with a localized environment that provides nutrients and/or soluble cell growth species to the cell or group of cells in a time-dependent manner and location-dependent concentrations. These conditions may represent growth conditions more like in vivo or alternatively allow for perturbation of these typical conditions to allow for research and growth under non-standard conditions. Standardized large-scale cell culture methods cannot be used to meet these requirements. However, there is a need for improved protocols to make the manipulation of one or several cells easier to: a) place the cell(s) into a microfluidic environment beneficial to support cell growth, viability, transplantability, or any combination thereof b) successfully maintaining the cell(s) and/or expanding the population of the cell(s); and/or c) defining the conditions that lead to successful growth and/or maintenance. The systems and methods described herein allow for more precise cell handling, environmental control, and cell isolation techniques for microfluidic cell culture, and can be used to generate, for example, germinal cell populations. This specification describes exemplary embodiments and applications of the invention. However, the invention is not limited to these exemplary embodiments and applications or to the manner in which these exemplary embodiments and applications operate or the manner in which they are described herein. Furthermore, the figures may show simplified or partial views, and the dimensions of elements in the figures may be exaggerated for clarity or otherwise may not be to scale. Additionally, when the terms "on", "attached to" or "coupled to" are used herein, an element (eg, a material, layer, a substrate, etc.) may be "on" another element, "Attached to" or "coupled to" another element, regardless of whether the element is directly on, directly attached to, or directly coupled to or between the element and the other element There are one or more intervening elements between the other element. Likewise, orientation (eg, above, below, top, bottom, side, up, down, below, above, up, down, horizontal, vertical, "x", "y", "z", etc.) (if provided ) are relative, for illustrative purposes only, for ease of illustration and discussion, and are in no way intended to be limiting. Additionally, when reference is made to a series of elements (eg, elements a, b, c), the reference is intended to include any of the listed elements themselves, any combination of not all of the listed elements, and/or combinations of one of all of the listed elements. The section divisions in this specification are for ease of inspection only and do not limit any combination of the elements discussed. As used herein, "substantially" means sufficient for the intended purpose. Thus, the term "substantially" allows for small, insignificant changes from an absolute or perfect state, dimension, measurement, result, or the like, such as would be expected by one of ordinary skill and would not significantly affect overall performance. "Substantially" means within 10% when used with a numerical value or parameter or property that can be expressed as a numerical value. The term "multiple" means more than one. As used herein, the term "plurality" can be 2, 3, 4, 5, 6, 7, 8, 9, 10, or more. As used herein, "air" refers to the composition of gases that predominate in the Earth's atmosphere. The four most abundant gases are nitrogen (typically present at a concentration of about 78% by volume, eg, in a range from about 70% to about 80%), oxygen (typically present at sea level at about 20.95% by volume, eg, in a range from about 10% to about 25%), argon (usually present at about 1.0% by volume, eg, in a range from about 0.1% to about 3%), and carbon dioxide (usually present at about 0.04%, For example, in a range from about 0.01% to about 0.07%). Air may have other trace gases such as methane, nitrous oxide or ozone, trace pollutants and organic materials such as pollen, diesel particles and the like. Air may contain water vapor (typically present at about 0.25%, or may be present in a range from about 10 ppm to about 5% by volume). Air can be provided for use in culture experiments as a filtered controlled composition and can be conditioned as described herein. As used herein, the term "positioning" is encompassed within its meaning "positioning." As used herein, a "microfluidic device" or "microfluidic device" includes a device configured to hold a fluid, one or more discrete microfluidic circuits, each microfluidic circuit being fluidly interconnected by Line elements (which include, but are not limited to) area(s), flow path(s), channel(s), chamber(s) and/or fence(s) and at least two ports configured to allow The fluid (and, as the case may be, tiny objects suspended in the fluid) flow into and/or out of the microfluidic device). Typically, a microfluidic circuit of a microfluidic device will comprise at least one microfluidic channel and at least one chamber, and will remain less than about 1 milliliter (eg, less than about 750 microliters, about 500 microliters, about 250 microliters, about 200 microliters, about 150 microliters, about 100 microliters, about 75 microliters, about 50 microliters, about 25 microliters, about 20 microliters, about 15 microliters, about 10 microliters, about 9 microliters, about 8 microliters, about 7 microliters, about 6 microliters, about 5 microliters, about 4 microliters, about 3 microliters or about 2 microliters) fluid volume. In certain embodiments, the microfluidic circuit holds about 1 microliter to about 2 microliters, about 1 microliter to about 3 microliters, about 1 microliter to about 4 microliters, about 1 microliter to about 5 microliters , about 2 microliters to about 5 microliters, about 2 microliters to about 8 microliters, about 2 microliters to about 10 microliters, about 2 microliters to about 12 microliters, about 2 microliters to about 15 microliters , about 2 microliters to about 20 microliters, about 5 microliters to about 20 microliters, about 5 microliters to about 30 microliters, about 5 microliters to about 40 microliters, about 5 microliters to about 50 microliters , about 10 microliters to about 50 microliters, about 10 microliters to about 75 microliters, about 10 microliters to about 100 microliters, about 20 microliters to about 100 microliters, about 20 microliters to about 150 microliters , about 20 microliters to about 200 microliters, about 50 microliters to about 200 microliters, about 50 microliters to about 250 microliters, or about 50 microliters to about 300 microliters. As used herein, a "nanofluidic device" or "nanofluidic device" is a type of microfluidic device having a microfluidic circuit containing a microfluidic circuit configured to hold less than about 1 microliter (eg, Less than about 750 nanoliters, about 500 nanoliters, about 250 nanoliters, about 200 nanoliters, about 150 nanoliters, about 100 nanoliters, about 75 nanoliters, about 50 nanoliters, about 25 nanoliters, about 20 nanoliters , about 15 nanoliters, about 10 nanoliters, about 9 nanoliters, about 8 nanoliters, about 7 nanoliters, about 6 nanoliters, about 5 nanoliters, about 4 nanoliters, about 3 nanoliters, about 2 nanoliters , about 1 nanoliter or less) at least one line element of a fluid volume. Typically, a nanofluidic device will include a plurality of circuit elements (eg, at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 , 50, 75, 100, 150, 200, 250, 300, 400, 500, 600, 700, 800, 900, 1000, 1500, 2000, 2500 , 3000, 3500, 4000, 4500, 5000, 6000, 7000, 8000, 9000, 10000 or more). In certain embodiments, one or more (eg, all) of the at least one wiring element is configured to hold about 100 picoliters to about 1 nanoliter, about 100 picoliters to about 2 nanoliters, about 100 picoliters to about 5 ng, from about 250 to about 2, about 250 to about 5, about 250 to about 10, about 500 to about 5, about 500 to about 10 ng, about 500 pic to about 15 ns, about 750 pic to about 10 ns, about 750 pic to about 15 ns, about 750 pic to about 20 ns, about 1 ns To a fluid volume of about 10 nanoliters, about 1 nanoliters to about 15 nanoliters, about 1 nanoliters to about 20 nanoliters, about 1 nanoliters to about 25 nanoliters, or about 1 nanoliters to about 50 nanoliters. In other embodiments, one or more (eg, all) of the at least one wiring element is configured to maintain about 100 nanoliters to about 200 nanoliters, about 100 nanoliters to about 300 nanoliters, about 100 nanoliters to about 100 nanoliters About 400 nanoliters, about 100 nanoliters to about 500 nanoliters, about 200 nanoliters to about 300 nanoliters, about 200 nanoliters to about 400 nanoliters, about 200 nanoliters to about 500 nanoliters, about 200 nanoliters to about 500 nanoliters about 600 na, about 200 to about 700 na, about 250 to about 400 na, about 250 to about 500 na, about 250 to about 600 na, or about 250 to about 250 to A fluid volume of about 750 nanoliters. As used herein, a "microfluidic channel" or "flow channel" refers to a flow region of a microfluidic device that has a length that is significantly longer than either the horizontal dimension or the vertical dimension. For example, the flow channel can be at least 5 times the length of the horizontal dimension or the vertical dimension, such as at least 10 times the length, at least 25 times the length, at least 100 times the length, at least 200 times the length, at least 300 times the length, at least 400 times the length, at least 500 times the length times longer or longer. In some embodiments, the length of a flow channel ranges from about 20,000 microns to about 100,000 microns, including any range therebetween. In some embodiments, the horizontal dimension is in the range from about 100 micrometers to about 1000 micrometers (eg, about 150 micrometers to about 500 micrometers) and the vertical dimension is from about 25 micrometers to about 200 micrometers (eg, from about 40 micrometers) to about 150 microns). It should be noted that a flow channel can have a variety of different spatial configurations in a microfluidic device and is therefore not limited to a completely linear element. For example, a flow channel can be or include sections having one or more of the following configurations: curvilinear, curved, helical, sloped, down sloped, bifurcated (eg, multiple different flow paths), and any combination thereof. Additionally, a flow channel may have different widening and contracting cross-sectional areas along its path to provide a desired fluid flow therein. As used herein, the term "obstruction" generally refers to a bump or similar type of structure that is large enough to partially (but not completely) obstruct the target micro-object between two different regions or circuit elements in a microfluidic device movement between. The two different regions/circuit elements can be, for example, a microfluidic incubation chamber and a microfluidic channel or a connecting region and an isolation region of a microfluidic incubation chamber. As used herein, the term "contraction" generally refers to a narrowing of one of the widths of a wiring element (or an interface between two wiring elements) in a microfluidic device. The constriction can be localized, for example, at the interface between a microfluidic incubation chamber and a microfluidic channel or at the interface between an isolation region and a connecting region of a microfluidic incubation chamber. As used herein, "transparent" refers to a material that allows visible light to pass through and does not substantially alter the light as it passes through. As used herein, the term "microscopic object" generally refers to any microscopic object that can be isolated and collected in accordance with the present invention. Non-limiting examples of tiny objects include: inanimate tiny objects, such as microparticles; microbeads (eg, polystyrene beads, Luminex™ beads, or the like); magnetic beads; microrods; microwires; quantum dots and the like The like; biological microscopic objects such as cells (eg embryonic cells, oocytes, sperm cells, cells dissociated from a tissue, eukaryotic cells, protist cells, animal cells, mammalian cells, human cells, immune cells (which Including, but not limited to, T cells, B cells, natural killer cells, macrophages, dendritic cells, and the like), fusion tumor cells, cultured cells, cells from a cell line, cancer cells (including (but not limited to) Not limited to) circulating tumor cells), infected cells, transfected and/or transformed cells (including but not limited to CHO cells), reporter cells, prokaryotic cells, and the like); biological organelles (e.g., cell nuclei) ); vesicles or complexes; synthetic vesicles; liposomes (e.g., synthetic or derived from membrane formulations); Methods Enzymol., 464:211-231)) and the like; or a combination of inanimate micro-objects and biological micro-objects (eg, microbeads attached to cells, liposome-coated microbeads, liposome-coated magnetic beads or the like). The beads may further have other moieties/molecules linked covalently or non-covalently, such as fluorescent labels, proteins, small molecule signaling moieties, antigens or chemical/biological species that can be used in an assay. As used herein, the term "cell" refers to a biological cell, which can be a plant cell, an animal cell (eg, a mammalian cell), a bacterial cell, a fungal cell, or the like. A mammalian cell can be derived, for example, from a human, a mouse, a rat, a horse, a goat, a sheep, a cow, a primate, or the like. A population of biological cells is "selective" if all viable cell lines in a population capable of reproduction are derived from the daughter cells of a single parent cell. The term "germ cell" refers to cells of the same colony. As used herein, a "colony" of biological cells refers to 2 or more cells (eg, 2 to 20, 4 to 40, 6 to 60, 8 to 80, 10 to 40) 100, 20 to 200, 40 to 400, 60 to 600, 80 to 800, 100 to 1000 or greater than 1000 cells). As used herein, the term "maintaining the cell(s)" refers to providing an environment that includes both fluid components and gaseous components, which provide the conditions necessary to keep the cells alive and/or expanded . As used herein, the term "expansion" when referring to a cell refers to an increase in the number of cells. As referred to herein, "breathable" means that a material or structure is permeable to at least one of oxygen, carbon dioxide, or nitrogen. In some embodiments, the breathable material or structure is permeable to more than one of oxygen, carbon dioxide, and nitrogen and further to all three of these gases. A "component" of a fluid medium is any chemical or biochemical molecule present in the medium, including solvent molecules, ions, small molecules, antibiotics, nucleotides and nucleosides, nucleic acids, amino acids, peptides, proteins , sugars, carbohydrates, lipids, fatty acids, cholesterol, metabolites or the like. As used herein with reference to a fluid medium, "diffusion" refers to the thermal movement of a component of the fluid medium along a concentration gradient. The phrase "flow of a medium" means the bulk movement of a fluid medium primarily due to any mechanism other than diffusion. For example, the flow of a medium may involve the movement of the fluid medium from one point to another (due to the pressure difference between the points). This flow may comprise a continuous, pulsed, periodic, random, intermittent or reciprocating flow of the liquid or any combination thereof. When one fluid medium flows into another fluid medium, turbulence and mixing of the medium can result. The phrase "substantially no flow" refers to a flow rate of a fluid medium that, when averaged over time, is less than the diffusion of a component of a material (such as an analyte of interest) into the fluid medium or the fluid medium rate within. The diffusion rate of the components of such a material can depend on, for example, the temperature, the size of the components, and the strength of the interaction between the components and the fluid medium. As used herein with reference to different regions within a microfluidic device, the phrase "fluidically connected" means that when the different regions are substantially filled with a fluid, such as a fluid medium, each of the regions The fluids are connected so as to form a single fluid. This does not mean that the fluids (or fluid medium) in these different regions need to have the same composition. Specifically, the fluids in the different fluidic connection regions of a microfluidic device can have different compositions (eg, different concentrations of solutes, such as proteins, carbohydrates, ions, or other molecules), which follow the solutes along their respective concentration gradients Movement and/or fluid flow through the device varies. A microfluidic (or nanofluidic) device may include "scanned" regions and "unscanned" regions. As used herein, a "sweep" region consists of one or more fluidic interconnection elements of a microfluidic circuit, each of which is in fluid flow through the microfluidic circuit experienced a flow of culture medium. The line elements that sweep a region can include, for example, regions, channels, and all or part of a chamber. As used herein, an "unswept" region consists of one or more fluidic interconnecting circuit elements of a microfluidic circuit, each of which is in fluid flow through the microfluidic circuit Substantially no fluid flux is experienced while the line. An unswept area can be fluidly connected to a swept area, provided that the fluid connections are structured to enable diffusion of the medium between the swept area and the unswept area but not substantially achieve medium in the swept area The flow between the swept area and the unswept area. Thus, the microfluidic device can be structured to substantially isolate an unswept region from the flow of one of the media in the swept region, while substantially only enabling diffusion between the swept and unswept regions Sexual fluid communication. For example, a flow channel of a microfluidic device is an example of a swept region, and an isolation region of a microfluidic device (described in further detail below) is an example of an unswept region. As used herein, a "non-swept" rate of fluid medium flow means sufficient to allow components of a second fluid medium in an isolated region of the growth chamber to diffuse into the first fluid medium in the flow region and/or or a flow rate that allows the components of the first fluid medium to diffuse into the second fluid medium in the isolated region; and wherein the first medium does not substantially flow further into the isolated region. As used herein, a "flow path" refers to one or more fluidically connecting line elements (eg, channel(s), region(s), chamber(s), and the like), which define a flow of a medium the trajectory and is subjected to a flow of the medium. Thus, a flow path is an example of a sweep area of a microfluidic device. Other circuit elements, such as unswept regions, may be fluidly connected to circuit elements that include the flow path but are not subject to flow of the medium of the flow path. As used herein, "arylene" refers to an aromatic group having 6 to 10 ring atoms (eg, C6-C10 aromatic or C6-C10 aryl) having at least one ring (which has a carbon A ring is a conjugated pi-electron system (eg, phenyl, perylene, and naphthyl) and has one or two points of attachment to other moieties of a molecule. Whenever a numerical range (such as "6 to 10") appears herein, it refers to each integer within the given range; for example, "6 to 10 ring atoms" means that an aryl group can consist of 6 rings atoms, 7 ring atoms, etc. (up to and including 10 ring atoms). The term includes monocyclic or fused-ring polycyclic (ie, adjacent pairs of rings that share ring atoms) groups. Examples of arylidene groups include, but are not limited to, phenylene groups, naphthylene groups, and the like). The arylidene moiety may be further substituted or may only have substitution at one or two points of attachment to the other moieties of the molecule. As used herein, "heteroaryl" refers to a 5- to 18-membered aromatic group (eg, C3-C13 heteroaryl) containing one or more ring heteroatoms selected from nitrogen, oxygen, and sulfur , and can include a monocyclic, bicyclic, tricyclic or tetracyclic ring system and the prefix "extended" indicates that the heteroaryl ring system has one or two points of attachment to other moieties of a molecule. Whenever a numerical range (such as "5 to 18") appears herein, it refers to each integer within the given range; for example, "5 to 18 ring atoms" means that the heteroaryl group may contain up to 5 Ring atoms, 6 ring atoms, etc. (up to and including 18 ring atoms). A "heteroaromatic" or "heteroaryl" moiety containing N refers to an aromatic group in which at least one of the skeletal atoms of the ring is a nitrogen atom. Polycyclic heteroaryl groups can be fused or non-fused. The heteroatom(s) in the heteroaryl group may optionally be oxidized. One or more nitrogen atoms, if present, may optionally be quaternized. A heteroaryl group is attached to the remainder of the molecule through any atom of the ring(s). Examples of heteroaryl include, but are not limited to, benzimidazolyl, phenylindolyl, isoxazolyl, thiazolyl, triazolyl, tetrazolyl, and thiophene (ie, thienyl group). The heteroaryl moiety may be further substituted or may have only one or two points of attachment to the other moieties of the molecule. As used herein, the term "heterocycle" refers to a saturated or partially unsaturated ring containing 1, 2 or 3, once or unsubstituted with 3, 4, 5, 6 or 7 members heteroatoms, preferably, containing 1 or 2 heteroatoms independently selected from oxygen, nitrogen and sulfur; or a bicyclic ring system containing up to 10 atoms containing atoms independently selected from oxygen, nitrogen and at least one heteroatom of sulfur, wherein the ring system containing the heteroatom is saturated. Examples of heterocyclyl include, but are not limited to, tetrahydrofuranyl, tetrahydrofurfuryl, pyrrolidinyl, hexahydropyridyl, 4-pyranyl, tetrahydropyranyl, thio, morpholinyl, hexahydro Pyrazinyl, dioxolanyl, dioxane, indolinyl and 5-methyl-6-chromanyl. Heterocyclyl groups have one or two points of attachment to other moieties of the molecule or may or may not be further substituted.systemThe present invention provides a system for culturing one or more biological cells in a microfluidic device comprising a microfluidic device comprising: a flow region configured to contain a first fluid medium a flow; and at least one growth chamber, wherein the growth chamber has at least one surface conditioned to support cell growth, viability, portability, or any combination thereof. Microfluidic devices and their use in operation and systems for observing such devices Figure 1 illustrates an example of a microfluidic device 100 and a system 150 that can be used in the practice of the present invention. A perspective view of the microfluidic device 100 is shown with its cover 110 partially cut away to provide a partial view of the microfluidic device 100 . Microfluidic device 100 generally includes a microfluidic circuit 120 that includes a flow path 106 through which a fluid medium 180 can flow to optionally deliver one or more tiny objects (not shown) to the microfluidic circuit 120 and/or through the microfluidic circuit 120. Although a single microfluidic circuit 120 is shown in FIG. 1, suitable microfluidic devices may include a plurality (eg, 2 or 3) of these microfluidic circuits. Regardless, the microfluidic device 100 can be configured as a nanofluidic device. In the embodiment depicted in FIG. 1 , the microfluidic circuit 120 includes a plurality of

microfluidic growth chambers

124 , 126 , 128 and 130 each having one or more openings in fluid communication with the flow path 106 . As will be discussed further below, the microfluidic growth chamber includes various features and structures that have been optimized to retain tiny objects in a microfluidic device, such as microfluidic device 100, even as a culture medium 180 flows through flow path 106 middle. However, before beginning to describe each of the above, a brief description of one of microfluidic device 100 and system 150 is provided. As generally depicted in FIG. 1 , the microfluidic circuit 120 is defined by an enclosure 102 . Although the enclosure 102 can be physically structured into different configurations, in the example shown in FIG. 1, the enclosure 102 is depicted as including a support structure 104 (eg, a substrate), a microfluidic circuit structure 108, and a lid 110. The support structure 104, the microfluidic circuit structure 108, and the cover 110 may be attached to each other. For example, the microfluidic circuit structure 108 can be disposed on an inner surface 109 of the support structure 104 , and the cover 110 can be disposed over the microfluidic circuit structure 108 . The microfluidic circuit structure 108 may together with the support structure 104 and the cover 110 define elements of the microfluidic circuit 120 . The support structure 104 can be located at the bottom of the microfluidic circuit 120 and the cover 110 can be located at the top of the microfluidic circuit 120, as shown in FIG. 1 . Alternatively, the support structure 104 and cover 110 may be configured in other orientations. For example, the support structure 104 can be located at the top of the microfluidic circuit 120 and the cover 110 can be located at the bottom of the microfluidic circuit 120 . Regardless, there may be one or more ports 107 each including a pathway into or out of the enclosure 102 . Examples of a passageway include a valve, a gate, a through hole, or the like. As shown, the port 107 is a through hole created by a gap in the microfluidic circuit structure 108 . However, the port 107 may be located in other components of the closure body 102, such as the cover 110. Although only one port 107 is shown in FIG. 1 , the microfluidic circuit 120 may have two or more ports 107 . For example, there may be a first port 107 that serves as an inlet for fluid into the microfluidic circuit 120 and a second port 107 that serves as an outlet for fluid to exit the microfluidic circuit 120 . Whether a port 107 acts as an inlet or an outlet may depend on the direction of fluid flow through the fluid path 106 . The support structure 104 may include one or more electrodes (not shown) and a substrate or interconnecting substrates. For example, the support structure 104 may include one or more semiconductor substrates, each of which is electrically connected to an electrode (eg, all or a subset of the semiconductor substrates may be electrically connected to a single electrode). The support structure 104 may further include a printed circuit board assembly ("PCBA"). For example, the semiconductor substrate(s) may be mounted on a PCBA. The microfluidic circuit structure 108 may define circuit elements of the microfluidic circuit 120 . Such circuit elements can include spaces or regions that can be fluidly interconnected when the microfluidic circuit 120 is filled with fluid, such as flow channels, chambers, fences, traps, and the like. In the microfluidic circuit 120 shown in FIG. 1 , the microfluidic circuit structure 108 includes a frame 114 and a microfluidic circuit material 116 . The frame 114 may partially or fully enclose the microfluidic circuit material 116 . Frame 114 may be, for example, a relatively rigid structure that substantially surrounds microfluidic circuit material 116 . For example, the frame 114 may comprise a metallic material. The microfluidic circuit material 116 may be patterned using voids or the like to define the circuit elements and interconnects of the microfluidic circuit 120 . The microfluidic circuit material 116 may comprise a flexible material that is breathable, such as a flexible polymer (eg, rubber, plastic, elastomer, polysiloxane, polydimethylsiloxane ("PDMS"), or the like By). Other examples of materials that may constitute the microfluidic circuit material 116 include molded glass, an etchable material such as polysilicon (eg, photo-patternable polysiloxane or "PPS"), photoresist (eg, SU8), or the like. The like. In some embodiments, these materials (and thus the microfluidic circuit material 116) may be rigid and/or may be substantially gas impermeable. Regardless, the microfluidic circuit material 116 may be disposed on the support structure 104 and Within frame 114. Cover 110 may be an integral part of frame 114 and/or microfluidic circuit material 116. Alternatively, cover 110 may be a different structural element, as depicted in Figure 1. Cover 110 may include the same as or A different material than the frame 114 and/or the microfluidic circuit material 116. Similarly, the support structure 104 may be a separate structure from the frame 114 or the microfluidic circuit material 116 (as shown in the figure) or be the frame 114 or microfluidic circuit material 116. An integral part of the fluidic circuit material 116. Likewise, the frame 114 and the microfluidic circuit material 116 may be separate structures (as shown in Figure 1) or may be integral parts of the same structure. In some embodiments, the cover 110 may include A rigid material. The rigid material may be glass or a material with similar properties. In some embodiments, the cover 110 may comprise a deformable material. The deformable material may be a polymer, such as PDMS. In some embodiments The lid 110 may include both rigid and deformable materials. For example, one or more portions of the lid 110 (eg, one or more portions positioned over the

growth chambers

124, 126, 128, 130) may include a The rigid material of 110 interfaces with a deformable material. In some embodiments, cover 110 may further include one or more electrodes. The one or more electrodes may include one or more that may be coated on glass or a similar insulating material Conductive oxide, such as indium tin oxide (ITO). Alternatively, the one or more electrodes may be flexible electrodes such as single-wall nanowires embedded in a deformable material such as a polymer (eg, PDMS) Clusters of nanotubes, multi-wall nanotubes, nanowires, conductive nanoparticles, or combinations thereof, etc. Flexible electrodes useful in microfluidic devices have been described, for example, in U.S. 2012/0325665 (Chiou et al.) , the contents of which are incorporated herein by reference. In some embodiments, cover 110 may be modified (eg, by adjusting all or part of a surface of microfluidic circuit 120 facing inward) to support cells Adhesion, viability and/or growth. The modification may include coating a synthetic or natural polymer. In some embodiments, the cover 110 and/or the support structure 104 may be light transmissive. The cover 110 may also include at least one Gas permeable material, such as PDMS or PPS. Figure 1 also shows a system 150 for operating and controlling a microfluidic device, such as microfluidic device 100. As depicted in the figure, system 150 includes a power source 192, an imaging device 194 and a tilt device 190. Power supply 192 may provide power to the microfluidic device 100 and/or tilt device 190. The device 190 is ramped to provide a bias voltage or current as desired. The power source 192 may, for example, include one or more alternating current (AC) and/or direct current (DC) voltage or current sources. Imaging device 194 may include a device for capturing images within microfluidic circuit 120, such as a digital camera. In some instances, imaging device 194 further includes a detector with a fast frame rate and/or high sensitivity (eg, for low light applications). Imaging device 194 may also include means for directing stimulation radiation and/or beams into microfluidic circuit 120 and collecting radiation and/or beams reflected or emitted from microfluidic circuit 120 (or the tiny objects contained therein). an institution. The emission beam may be in the visible spectrum and may, for example, comprise fluorescent emission. The reflected beam may comprise reflected emission from an LED or a broad spectrum lamp, such as a mercury lamp (eg, a high pressure mercury lamp) or a xenon arc lamp. As discussed with respect to FIG. 3, the imaging device 194 may further include a microscope (or a string of optical elements), which may or may not include an eyepiece. System 150 may further include a tilt device 190 configured to rotate a microfluidic device 100 about one or more axes of rotation. In some embodiments, the tilt device 190 is configured to support and/or retain the enclosure 102 including the microfluidic circuit 120 about at least one axis such that the microfluidic device 100 (and thus the microfluidic circuit 120) can be maintained in a horizontal orientation (ie, 0° with respect to the x-axis and y-axis), a vertical orientation (ie, 90° with respect to the x-axis and/or the y-axis), or any orientation in between. The orientation of microfluidic device 100 (and microfluidic circuit 120 ) relative to an axis is referred to herein as the "tilt angle" of microfluidic device 100 (and microfluidic circuit 120 ). For example, tilting device 190 may tilt microfluidic device 100 relative to the x-axis by 0.1°, 0.2°, 0.3°, 0.4°, 0.5°, 0.6°, 0.7°, 0.8°, 0.9°, 1°, 2°, 3° °, 4°, 5°, 10°, 15°, 20°, 25°, 30°, 35°, 40°, 45°, 50°, 55°, 60°, 65°, 70°, 75°, 80°, 90° or any degree in between. The horizontal orientation (and thus the x- and y-axes) is defined as normal to a vertical axis defined by gravity. Tilt the device can also tilt the microfluidic device 100 (and the microfluidic circuit 120 ) by any degree greater than 90° with respect to the x-axis and/or the y-axis, or tilt the microfluidic device 100 (and the microfluidic circuit 120 ) with respect to the x-axis Or the y-axis is tilted 180° so that the microfluidic device 100 (and the microfluidic circuit 120) is completely upside down. Similarly, in some embodiments, tilting device 190 tilts microfluidic device 100 (and microfluidic circuit 120 ) about an axis of rotation defined by flow path 106 or some other portion of microfluidic circuit 120 . In some instances, microfluidic device 100 is tilted into a vertical orientation such that flow path 106 is positioned above one or more growth chambers or below one or more growth chambers. As used herein, the term "above" indicates that, on a vertical axis defined by gravity, the flow path 106 is positioned higher than one or more growth chambers (ie, one of the growth chambers above a flow path 106 The object will have a gravitational potential energy that is one higher than one of the objects in the flow path). As used herein, the term "below" indicates that, on a vertical axis defined by gravity, the flow path 106 is positioned below one or more growth chambers (ie, one of the growth chambers below a flow path 106 The object will have one lower gravitational potential energy than one of the objects in the flow path). In some instances, tilting device 190 tilts microfluidic device 100 about an axis parallel to flow path 106 . Also, the microfluidic device 100 can be tilted at an angle of less than 90° such that the flow path 106 is positioned above or below the growth chamber or chambers and not directly above or below the growth chambers. In other instances, tilting device 190 tilts microfluidic device 100 about an axis perpendicular to flow path 106 . In other instances, tilting device 190 tilts microfluidic device 100 about an axis that is neither parallel to flow path 106 nor perpendicular to flow path 106 . System 150 may further include a culture medium source 178 . The medium source 178 (eg, a container, reservoir, or the like) may include multiple segments or containers each for holding a different fluid medium 180 . Thus, the medium source 178 may be a device located outside the microfluidic device 100 and separate from the microfluidic device 100, as depicted in FIG. 1 . Alternatively, the medium source 178 may be located in whole or in part within the enclosure 102 of the microfluidic device 100 . For example, medium source 178 may include a reservoir, which is part of microfluidic device 100 . FIG. 1 also shows a simplified block diagram depiction of an example of control and monitoring apparatus 152 that forms part of system 150 and that may be used with a microfluidic device 100 . As shown in the figure, examples of such control and monitoring equipment 152 include: a master controller 154 including: a medium module 160 for controlling a medium source 178 , a medium module 160 for controlling microfluidic circuits 120 ( not shown) and/or movement and/or selection of medium (eg, droplets of medium), a power module 162 for controlling an imaging device 194 (eg, a camera, microscope, light source, or any combination thereof) ) to capture images (eg, digital images), an imaging module 164 , and a tilt module 166 for controlling a tilt device 190 . The control apparatus 152 may also include other modules 168 for controlling, monitoring, or performing other functions with respect to the microfluidic device 100 . As shown in the figure, apparatus 152 may further include a display device 170 and an input/output device 172 . The main controller 154 may include a control module 156 and a digital memory 158 . The control module 156 may include, for example, a digital processor configured to operate according to machine-executable instructions (eg, software, firmware, source code or the like) stored in the memory 158 as non-transitory data or signals. similar) to operate. Alternatively or additionally, the control module 156 may include hardwired digital circuits and/or analog circuits. Media module 160, power module 162, imaging module 164, tilt module 166, and/or other modules 168 may be similarly configured. Thus, by any one or more of the main controller 154, media module 160, power module 162, imaging module 164, tilt module 166, and/or other modules 168 configured as discussed above, such as The functions, procedures, acts, actions, or steps of a procedure discussed herein are performed with respect to the microfluidic device 100 or any other microfluidic device. Similarly, main controller 154, media module 160, power module 162, imaging module 164, tilt module 166, and/or other modules 168 may be communicatively coupled to transmit and receive for the purposes discussed herein Information in any function, procedure, action, action or step. The medium module 160 controls the medium source 178 . For example, the medium module 160 can control the medium source 178 to deliver a selected fluid medium 180 into the enclosure 102 (eg, through an inlet 107). Media module 160 can also control the removal of media from enclosure 102 (eg, through an outlet (not shown)). Thus, one or more culture media can be selectively input into and removed from the microfluidic circuit 120 . The medium module 160 can also control the flow of the fluid medium 180 in the flow path 106 within the microfluidic circuit 120 . For example, in some embodiments, the medium module 160 stops the flow of the medium 180 in the flow path 106 and stops the flow of the medium 180 through the enclosure before the tilt module 166 causes the tilt device 190 to tilt the microfluidic device 100 by a desired tilt angle 102. The power module 162 can be configured to control the selection, capture, and movement of tiny objects (not shown) in the microfluidic circuit 120 . As will be discussed below with respect to FIGS. 2A and 2B , enclosure 102 may include a dielectrophoretic (DEP), optoelectronic tweezers (OET), and/or optoelectronic wetting (OEW) configuration (not shown in FIG. 1 ), and kinetic mode Group 162 can control activation electrodes and/or transistors (eg, phototransistors) to select and move flow path 106 and/or tiny objects (not shown) and/or media in

growth chambers

124, 126, 128, 130 droplets (not shown). The imaging module 164 can control the imaging device 194 . For example, imaging module 164 may receive and process image data from imaging device 194 . Image data from imaging device 194 may include any type of information captured by imaging device 194 (eg, presence or absence of microscopic objects, droplets of culture medium, accumulation of labels such as fluorescent labels, etc.). Imaging module 164 may use the information captured by imaging device 194 to further calculate the position of objects (eg, tiny objects, droplets of culture medium) and/or the velocity of motion of such objects within microfluidic device 100 . The tilting module 166 can control the tilting movement of the tilting device 190 . Alternatively or additionally, the tilt module 166 can control the tilt rate and timing to optimize the transfer of microscopic objects to one or more growth chambers via gravity. Tilt module 166 is communicatively coupled with imaging module 164 to receive data describing the motion of microscopic objects and/or droplets of culture medium in microfluidic circuit 120 . The tilt module 166 can use this data to adjust the tilt angle of the microfluidic circuit 120 to adjust the rate of movement of tiny objects and/or droplets of culture medium in the microfluidic circuit 120 . Tilt module 166 may also use this data to iteratively adjust the position of a tiny object and/or drop of culture medium in microfluidic circuit 120 . In the example shown in FIG. 1 , the microfluidic circuit 120 is shown to include a microfluidic channel 122 and

growth chambers

124 , 126 , 128 , 130 . Each chamber includes an opening to the channel 122, but other portions are closed such that the chamber can substantially isolate the microscopic objects within the chamber from the fluid medium 180 and/or microscopic objects in the flow path 106 of the channel 122 or in other chambers. In some embodiments,

chambers

124 , 126 , 128 , 130 are configured to physically enclose one or more tiny objects within microfluidic circuit 120 . Growth chambers in accordance with the present invention may include various shapes, surfaces and features optimized for use with DEP, OET, OEW and/or gravity, as will be discussed and shown in detail below. Microfluidic circuit 120 may include any number of microfluidic growth chambers. Although 5 growth chambers are shown, the microfluidic circuit 120 may have fewer or more growth chambers. In some embodiments, the microfluidic circuit 120 includes a plurality of microfluidic growth chambers, wherein two or more of the growth chambers include different structures and/or features. In the embodiment depicted in FIG. 1, a single channel 122 and flow path 106 are shown. However, other embodiments may contain channels 122 each configured to include a flow path 106 . The microfluidic circuit 120 further includes an inlet valve or inlet 107 in fluid communication with the flow path 106 and the fluid medium 180 , whereby the fluid medium 180 can enter the channel 122 via the inlet 107 . In some instances, flow path 106 includes a single path. In some instances, the single path is configured in a zigzag pattern whereby the flow path 106 travels across the microfluidic device 100 two or more times in alternating directions. In some instances, the microfluidic circuit 120 includes a plurality of parallel channels 122 and flow paths 106, wherein the fluid medium 180 within each flow path 106 flows in the same direction. In some instances, the fluid medium within each flow path 106 flows in at least one of a forward direction or an opposite direction. In some instances, a plurality of growth chambers are configured (eg, with respect to a channel 122) such that they can be loaded with target micro-objects in parallel. In some embodiments, the microfluidic circuit 120 further includes one or more fine object traps 132 . Trap 132 is generally formed in a wall forming a boundary of a channel 122 and may be positioned to open with respect to one or more of

microfluidic growth chambers

124 , 126 , 128 , 130 . In some embodiments, trap 132 is configured to receive or capture a single tiny object from one of flow path 106 . In some embodiments, trap 132 is configured to receive or capture a plurality of tiny objects from flow path 106 . In some instances, trap 132 includes a volume approximately equal to the volume of a single target micro-object. The trap 132 may further include an opening in the trap 132 that is configured to facilitate the flow of target fines to the trap 132 . In some instances, trap 132 includes an opening having a height and width approximately equal to the dimensions of a single target micro object, thereby preventing larger micro objects from entering the micro object trap. The trap 132 may further include other features that are configured to facilitate retention of the target micro-objects within the trap 132 . In some instances, the trap 132 is aligned with and on the side of a channel 122 opposite the opening of a microfluidic growth chamber such that the microfluidic device 100 surrounds parallel to the channel After one axis of 122 is tilted, the captured tiny objects exit the trap 132 on a trajectory that causes the tiny objects to fall into the opening of the growth chamber. In some instances, the trap 132 includes a side passage 134 that is smaller than the target micro object to facilitate flow through the trap 132 and thereby increase the likelihood of capturing one of the micro objects in the trap 132 . In some embodiments, a dielectrophoretic (DEP) force is applied across the fluid medium 180 (eg, in the flow path and/or in the growth chamber) via one or more electrodes (not shown) for manipulation, transport , separates and sorts tiny objects located in the flow path and/or growth chamber. For example, in some embodiments, a DEP force is applied to one or more portions of the microfluidic circuit 120 to transfer a single tiny object from the flow path 106 into a desired microfluidic growth chamber. In some embodiments, the DEP force is used to prevent a tiny object within a growth chamber (eg,

growth chamber

124, 126, 128, or 130) from being expelled from the growth chamber. Further, in some embodiments, the DEP force is used to selectively remove a tiny object previously collected in accordance with the teachings of the present invention from a growth chamber. In some embodiments, the DEP force includes an optoelectronic tweezers (OET) force. In other embodiments, an electro-optical wetting (OEW) force is applied to one or more of the support structure 104 (and/or lid 110 ) of the microfluidic device 100 via one or more electrodes (not shown) Locations, such as those that help define flow paths and/or growth chambers, to manipulate, transport, separate, and sort droplets positioned in microfluidic circuit 120 . For example, in some embodiments, an OEW force is applied to one or more locations in support structure 104 (and/or lid 110) to transfer a single droplet from flow path 106 into a desired microfluidic growth chamber. In some embodiments, the OEW force is used to prevent a droplet within a growth chamber (eg,

growth chamber

124, 126, 128, or 130) from being expelled from the growth chamber. Further, in some embodiments, the OEW force is used to selectively remove a droplet previously collected in accordance with the teachings of the present invention from a growth chamber. In some embodiments, DEP forces and/or OEW forces are combined with other forces, such as flow forces and/or gravity, to manipulate, transport, separate, and sort tiny objects and/or droplets within microfluidic circuit 120 . For example, enclosure 102 can be tilted (eg, by tilting device 190) to position flow path 106 and tiny objects positioned within flow path 106 above the microfluidic growth chamber, and gravity can tilt the tiny objects and/or liquid Drops are transported into the chamber. In some embodiments, DEP and/or OEW forces may be applied before other forces. In other embodiments, the DEP force and/or the OEW force may be applied after the other force. In other examples, the DEP force and/or the OEW force may be applied simultaneously with the other force or in an alternating manner with the other force. 2A-2F illustrate various embodiments of microfluidic devices that may be used in the practice of the present invention. Figure 2A depicts one embodiment in which the microfluidic device 200 is configured as an optically actuated electrodynamic device. Various optically actuated electrodynamic devices are known in the art, including devices having an optoelectronic tweezers (OET) configuration and devices having an optoelectronic wetting (OEW) configuration. Examples of suitable OET configurations are depicted in the following US patent documents, each of which is incorporated herein by reference in its entirety: US Patent No. RE 44,711 (Wu et al.) (originally published as US Patent No. 7,612,355 No.) and US Patent No. 7,956,339 (Ohta et al.). Examples of OEW configurations are depicted in US Patent No. 6,958,132 (Chiou et al.) and US Patent Application Publication No. 2012/0024708 (Chiou et al.), which are incorporated herein by reference in their entirety. Yet another example of an optically actuated electrodynamic device including a combined OET/OEW configuration, US Patent Publication Nos. 20150306598 (Khandros et al.) and 20150306599 (Khandros et al.) and their corresponding PCT publications WO2015/ Examples of OET/OEW configurations of this combination are shown in 164846 and WO2015/164847, all of which are incorporated herein by reference in their entirety.Dynamic Microfluidic Device Configuration As described above, the control and monitoring equipment of the system may include a power module for selecting and moving objects, such as tiny objects or droplets, in the microfluidic circuit of a microfluidic device. Microfluidic devices can have various dynamic configurations depending on the type of object being moved and other considerations. For example, a dielectrophoretic (DEP) configuration can be used to select and move tiny objects in a microfluidic circuit. Accordingly, the support structure 104 and/or the lid 110 of the microfluidic device 100 may include a DEP configuration for selectively inducing DEP forces on microscopic objects in a fluid medium 180 in a microfluidic circuit 120 and thereby select , capture and/or move individual tiny objects or groups of tiny objects. Alternatively, the support structure 104 and/or lid 110 of the microfluidic device 100 may include an EW for selectively inducing electrowetting (EW) forces on droplets in a fluidic medium 180 in a microfluidic circuit 120 Configure and thereby select, capture and/or move individual droplets or groups of droplets. An example of a microfluidic device 200 including a DEP configuration is shown in FIGS. 2A and 2B. 2A and 2B show, respectively, a side cross-sectional view and a top cross-sectional view of a portion of an enclosure 102 of a microfluidic device 200 having an open region/chamber 202, it should be understood that Region/chamber 202 may be part of a fluid circuit element having a more detailed structure, such as a growth chamber, a flow region, or a flow channel. Additionally, the microfluidic device 200 may contain other fluidic circuit elements. For example, microfluidic device 200 may include a plurality of growth chambers and/or one or more flow regions or flow channels, such as the growth chambers and/or flow regions or flow channels described herein with respect to microfluidic device 100 . A DEP configuration can be incorporated into any of these fluidic circuit elements of microfluidic device 200 or selected portions thereof. It should be further appreciated that any of the microfluidic device components and system components described above or to be described below may be incorporated into and/or used with microfluidic device 200 . For example, the system 150 including the control and monitoring apparatus 152 described above can be combined with a microcomputer including one or more of the media module 160 , the power module 162 , the imaging module 164 , the tilt module 166 , and the other modules 168 . The fluidic device 200 is used together. As seen in FIG. 2A, the microfluidic device 200 includes: a support structure 104 having a bottom electrode 204 and an electrode activation substrate 206 overlying the bottom electrode 204; and a lid 110 having a top electrode 210, Wherein the top electrode 210 is spaced apart from the bottom electrode 204 . Top electrode 210 and electrode activation substrate 206 define opposing surfaces of region/chamber 202 . Thus, a medium 180 contained in the region/chamber 202 provides a resistive connection between the top electrode 210 and the electrode activation substrate 206 . Also shown is a power supply 212 configured to connect to the bottom electrode 204 and the top electrode 210 and to generate a bias voltage between the electrodes required to generate the DEP force in the region/chamber 202. The power source 212 can be, for example, an alternating current (AC) power source. In certain embodiments, the microfluidic device 200 depicted in FIGS. 2A and 2B may have an optically actuated DEP configuration. Accordingly, changing the light pattern 222 from the light source 220 , which can be controlled by the power module 162 , can selectively activate and deactivate changing the pattern of DEP electrodes at the region 214 of the inner surface 208 of the electrode activation substrate 206 . (Hereinafter, the region 214 of a microfluidic device having a DEP configuration is referred to as the "DEP electrode region"). As depicted in FIG. 2B, a light pattern 222 directed onto the inner surface 208 of the electrode activation substrate 206 may illuminate the selected DEP electrode region 214a (shown in white) in a pattern, such as a square. The unirradiated DEP electrode region 214 (cross-hatched) is hereinafter referred to as the "dark" DEP electrode region 214 . The relative electrical impedance through the DEP electrode activation substrate 206 (ie, from the bottom electrode 204 to the inner surface 208 of the electrode activation substrate 206 that interfaces with the culture medium 180 in the flow region 106 ) is greater than the area through each dark DEP electrode area 214/ The relative electrical impedance of the culture medium 180 in the chamber 202 (ie, from the inner surface 208 of the electrode activation substrate 206 to the top electrode 210 of the lid 110). However, an irradiated DEP electrode area 214a exhibits a reduced relative impedance through one of the electrode activation substrates 206 that is less than the relative impedance through the medium 180 in the area/chamber 202 at each irradiated DEP electrode area 214a. After activation of the power source 212, the aforementioned DEP configuration creates an electric field gradient in the fluid medium 180 between the irradiated DEP electrode region 214a and the adjacent dark DEP electrode region 214, which in turn induces attraction or repulsion to adjacent ones in the fluid medium 180 Local DEP force for tiny objects (not shown). Thus, attraction can be selectively activated and deactivated at many different DEP electrode regions 214 at the inner surface 208 of the region/chamber 202 by changing the light pattern 222 projected from a light source 220 into the microfluidic device 200 Or a DEP electrode that repels tiny objects in the fluid medium 180 . Whether the DEP force attracts or repels nearby micro-objects may depend on parameters such as the frequency of the power source 212 and the dielectric properties of the culture medium 180 and/or micro-objects (not shown). The square pattern 224 of irradiated DEP electrode regions 214a depicted in FIG. 2B is only one example. Any pattern of DEP electrode regions 214 may be illuminated (and thereby activated) by light pattern 222 projected into device 200, and the pattern of illuminated/activated DEP electrode regions 214 may be repeatedly changed by changing or moving light pattern 222 . In some embodiments, the electrode activation substrate 206 may include or consist of a photoconductive material. In such embodiments, the inner surface 208 of the electrode activation substrate 206 may be featureless. For example, the electrode activation substrate 206 may include or consist of a layer of hydrogenated amorphous silicon (a-Si:H). a-Si:H may include, for example, about 8% to about 40% hydrogen (calculated as 100*number of hydrogen atoms/total number of hydrogen atoms and silicon atoms). The a-Si:H layer may have a thickness of about 500 nanometers to about 2.0 micrometers. In these embodiments, the DEP electrode regions 214 can be created anywhere on the inner surface 208 of the electrode activation substrate 206 and in any pattern according to the light pattern 222 on the inner surface 208 of the electrode activation substrate 206 . Therefore, the number and pattern of the DEP electrode regions 214 need not be fixed, but may correspond to the light pattern 222 . Examples of microfluidic devices having a DEP configuration including a photoconductive layer such as the one discussed above have been described, for example, in US Patent No. RE 44,711 (Wu et al.) (originally published as US Patent No. 7,612,355) , the entire contents of which are incorporated herein by reference. In other embodiments, the electrode activation substrate 206 may comprise a substrate including a plurality of doped layers, electrically insulating layers (or regions) and conductive layers forming semiconductor integrated circuits, such as is known in the semiconductor art. For example, electrode activation substrate 206 may include a plurality of phototransistors (including, for example, lateral bipolar phototransistors), each phototransistor corresponding to a DEP electrode region 214 . Alternatively, the electrode activation substrate 206 may include electrodes (eg, conductive metal electrodes) controlled by phototransistor switches, wherein each such electrode corresponds to a DEP electrode region 214 . Electrode activation substrate 206 may include a pattern of these phototransistors or phototransistor control electrodes. For example, the pattern may be an array of substantially square phototransistors or phototransistor control electrodes arranged in columns and rows, such as shown in Figure 2B. Alternatively, the pattern may be an array of substantially hexagonal phototransistors or phototransistor control electrodes forming a hexagonal lattice. Regardless of the pattern, the circuit elements can form electrical connections between the DEP electrode regions 214 at the inner surface 208 of the electrode activation substrate 206 and the bottom electrode 210 , and these electrical connections (ie, phototransistors or electrodes) can be selected by the light pattern 222 to activate and deactivate automatically. When not activated, each electrical connection can have a high impedance such that the inner surface 208 of the substrate 206 is activated by the electrode (ie, from the bottom electrode 204 to the electrode that interfaces with the culture medium 180 in the region/chamber 202 ) ) is greater than the relative impedance through the medium 180 at the corresponding DEP electrode region 214 (ie, from the inner surface 208 of the electrode activation substrate 206 to the top electrode 210 of the lid 110). However, when activated by light in the light pattern 222, the relative impedance of the substrate 206 through the electrode activation is less than the relative impedance through the medium 180 at each irradiated DEP electrode area 214, thereby activating the DEP electrode at the corresponding DEP electrode area 214 , as discussed above. Thus, electrodes in the region/chamber 202 can be selectively activated and deactivated at many different DEP electrode regions 214 at the inner surface 208 of the substrate 206 in a manner determined by the light pattern 222 to attract or repel the electrodes in the medium 180. DEP electrodes for tiny objects (not shown). Examples of microfluidic devices having electrode-activated substrates including phototransistors have been described, for example, in US Pat. No. 7,956,339 (Ohta et al.) (see, for example, device 300 depicted in FIGS. 21 and 22 and descriptions thereof), The entire contents of this patent are incorporated herein by reference. Examples of microfluidic devices having electrode-activated substrates including electrodes controlled by phototransistor switches have been described, for example, in US Patent Application No. 2014/0124370 (Short et al.) (see, eg, those depicted throughout the drawings).

devices

200, 400, 500, 600, and 900 and their descriptions), which are incorporated herein by reference in their entirety. In some embodiments of microfluidic devices in a DEP configuration, the top electrode 210 is part of a first wall (or lid 110 ) of the enclosure 102 and the electrode activation substrate 206 and bottom electrode 204 are one of the enclosures 102 Portion of the second wall (or support structure 104). The region/chamber 202 may be interposed between the first wall and the second wall. In other embodiments, electrode 210 is part of the second wall (or support structure 104) and one or both of the electrode activation substrate 206 and/or electrode 210 is part of the first wall (or cover 110). Also, the light source 220 may alternatively be used to illuminate the enclosure 102 from below. For the microfluidic device 200 of FIGS. 2A-2B having a DEP configuration, the power module 162 may activate a pattern (eg, a square pattern 224 ) by projecting a light pattern 222 into the device 200 (which surrounds the And the electrodes in a micro-object (not shown) in the culture medium 180 in the trapped region/chamber 202 activate a first set of one or more DEPs at the DEP electrode region 214a of the inner surface 208 of the substrate 206 electrode to select the tiny object. The power module 162 can then move the captured tiny objects by moving the light pattern 222 relative to the device 200 to activate a second set of one or more DEP electrodes at the DEP electrode region 214 . Alternatively, device 200 may be moved relative to light pattern 222 . In other embodiments, the microfluidic device 200 may have a DEP configuration that does not rely on light activation of DEP electrodes at the inner surface 208 of the electrode activation substrate 206 . For example, electrode activation substrate 206 may include selectively addressable and energized electrodes positioned relative to a surface that includes at least one electrode (eg, lid 110). A switch, such as a transistor switch in a semiconductor substrate, can be selectively turned on and off to activate or deactivate the DEP electrode at the DEP electrode region 214, thereby creating an effect in the region/chamber 202 near the activation DEP electrode A net DEP force for a tiny object (not shown). Depending on characteristics such as the frequency of the power source 212 and the medium (not shown) and/or the dielectric properties of the tiny objects in the region/chamber 202, the DEP force can attract or repel an adjacent tiny object. One or more tiny objects in the region/chamber 202 can be trapped and made available by selectively activating and deactivating a set of DEP electrodes (eg, at a set of DEP electrode regions 214 forming a square pattern 224). Move within the area/chamber 202 . The power module 162 in FIG. 1 can control these switches and thus activate and deactivate individual DEP electrodes to select, trap and move certain tiny objects (not shown) around the area/chamber 202 . Microfluidic devices having a DEP configuration comprising selectively addressable and energized electrodes are known in the art and have been described, for example, in US Pat. Nos. 6,294,063 (Becker et al.) and 6,942,776 (Medoro) , the entire contents of these patents are incorporated herein by reference. As yet another example, the microfluidic device 200 can have an electrowetting (EW) configuration that can replace the DEP configuration or can be positioned in a portion of the microfluidic device 200 that is separate from the portion having the DEP configuration. The EW configuration can be an opto-wetting configuration or a dielectric wetting (EWOD) configuration, both of which are known in the art. In some EW configurations, the support structure 104 has an electrode activation substrate 206 sandwiched between a dielectric layer (not shown) and the bottom electrode 204 . The dielectric layer may include a hydrophobic material and/or may be coated with a hydrophobic material. For the microfluidic device 200 having an EW configuration, the inner surface 208 of the support structure 104 is the inner surface of the dielectric layer or its hydrophobic coating. The dielectric layer (not shown) may include one or more oxide layers and may have a thickness of about 50 nm to about 250 nm (eg, about 125 nm to about 175 nm). In certain embodiments, the dielectric layer may include a layer of oxide, such as a metal oxide (eg, aluminum oxide or hafnium oxide). In some embodiments, the dielectric layer may include a dielectric material other than a metal oxide, such as silicon oxide or nitride. Regardless of the exact composition and thickness, the dielectric layer can have an impedance of about 10 kiloohms to about 50 kiloohms. In some embodiments, the surface of the dielectric layer facing inwardly of the region/chamber 202 is coated with a hydrophobic material. The hydrophobic material may include, for example, fluorinated carbon molecules. Examples of fluorocarbon molecules include perfluoropolymers such as polytetrafluoroethylene (eg, TEFLON®) or poly(2,3-difluoromethylene-perfluorotetrahydrofuran) (eg, CYTOP™). The molecules that make up the hydrophobic material can be covalently bonded to the surface of the dielectric layer. For example, the molecules of the hydrophobic material can be covalently bonded to the surface of the dielectric layer via a linker such as a siloxane, phosphonate or thiol group. Thus, in some embodiments, the hydrophobic material may comprise an alkyl-terminated siloxane, an alkyl-terminated phosphonic acid, or an alkyl-terminated thiol. Alkyl groups can be long chain hydrocarbons (eg, having at least 10 carbons or a chain of at least 16, 18, 20, 22, or more carbons). Alternatively, fluorinated (or perfluorinated) carbon chains can be used to replace the alkyl groups. Thus, for example, the hydrophobic material may comprise a fluoroalkyl terminated siloxane, a fluoroalkyl terminated phosphonic acid, or a fluoroalkyl terminated thiol. In some embodiments, the hydrophobic coating has a thickness of one of about 10 nanometers to about 50 nanometers. In other embodiments, the hydrophobic coating has a thickness of less than 10 nanometers (eg, less than 5 nanometers or about 1.5 nanometers to about 3.0 nanometers). In some embodiments, the lid 110 of a microfluidic device 200 having an electrowetting configuration is also coated with a hydrophobic material (not shown). The hydrophobic material can be the same hydrophobic material used to coat the dielectric layer of the support structure 104 , and the hydrophobic coating can have a thickness that is substantially the same as the thickness of the hydrophobic coating on the dielectric layer of the support structure 104 . Furthermore, the lid 110 may include an electrode activation substrate 206 sandwiched between a dielectric layer and the top electrode 210 by way of the support structure 104 . The electrode activation substrate 206 and the dielectric layers of the lid 110 may have the same composition and/or dimensions as the electrode activation substrate 206 and the dielectric layers of the support structure 104 . Thus, the microfluidic device 200 can have two electrowetting surfaces. In some embodiments, the electrode activation substrate 206 may include a photoconductive material, such as described above. Accordingly, in some embodiments, the electrode activation substrate 206 may comprise or consist of a layer of hydrogenated amorphous silicon (a-Si:H). For example, a-Si:H may include about 8% to 40% hydrogen (calculated as 100*number of hydrogen atoms/total number of hydrogen atoms and silicon atoms). The a-Si:H layer may have a thickness of about 500 nanometers to about 2.0 micrometers. Alternatively, the electrode activation substrate 206 may include electrodes (eg, conductive metal electrodes) controlled by phototransistor switches, as described above. Electrode-activated substrates known in the art are known and/or can be used to construct microfluidic devices having an optoelectronic wetting configuration. For example, US Pat. No. 6,958,132 (Chiou et al.), which is incorporated herein by reference in its entirety, discloses opto-wetting configurations with a photoconductive material such as a-Si:H, while the above Referenced US Patent Publication No. 2014/0124370 (Short et al.) discloses electrode activation substrates having electrodes controlled by phototransistor switches. Thus, the microfluidic device 200 can have a photowetting configuration, and the light pattern 222 can be used to activate the photoconductive EW region or the photoresponsive EW electrode in the electrode activation substrate 206 . These activated EW regions or EW electrodes of the electrode activation substrate 206 can generate an electrowetting force at the inner surface 208 of the support structure 104 (ie, the inner surface of the overlying dielectric layer or its hydrophobic coating). A droplet (eg, which contains an aqueous medium, a solution, or a solution containing an aqueous medium, a solution) can be brought into contact with the inner surface 208 of the support structure 104 by changing the light pattern 222 incident on the electrode activation substrate 206 (or moving the microfluidic device 200 relative to the light source 220). or solvent) move through an immiscible fluid (eg, an oil medium) present in the region/chamber 202. In other embodiments, the microfluidic device 200 may have an EWOD configuration, and the electrode activation substrate 206 may include selectively addressable and energized electrodes independent of the light used for activation. Thus, electrode activation substrate 206 may include a pattern of these electrowetting (EW) electrodes. For example, the pattern may be an array of substantially square EW electrodes configured in columns and rows, such as shown in Figure 2B. Alternatively, the pattern may be an array of substantially hexagonal EW electrodes forming a hexagonal lattice. Regardless of the pattern, the EW electrodes can be selectively activated (or deactivated) by electrical switches, such as transistor switches in a semiconductor substrate. Droplets (not shown) contacting the inner surface 208 of the overlying dielectric layer or its hydrophobic coating can be brought into the region/chamber 202 by selectively activating and deactivating the electrodes to activate the EW electrodes in the substrate 206 move. The power module 162 in FIG. 1 can control these switches and thus activate and deactivate individual EW electrodes to select and move specific droplets around the region/chamber 202. Microfluidic devices having an EWOD configuration with selectively addressable and energized electrodes are known in the art and are described, for example, in US Pat. No. 8,685,344 (Sundarsan et al.), the entirety of which The contents are incorporated herein by reference. Regardless of the configuration of the microfluidic device 200, a power supply 212 may be used to provide a potential (eg, an AC voltage potential) to power the circuitry of the microfluidic device 200. Power supply 212 may be the same as power supply 192 referenced in FIG. 1 or may be a component of power supply 192 referenced in FIG. 1 . Power supply 212 may be configured to provide an AC voltage and/or current to top electrode 210 and bottom electrode 204 . For an AC voltage, the power supply 212 can provide a range of frequencies and a range of average or peak power (eg, voltage or current) sufficient to generate a net DEP force (or electrowetting force) strong enough to trap and move the region/chamber Tiny objects (not shown) in 202 (as discussed above) and/or changing the inner surface 208 of support structure 104 in region/chamber 202 (ie, the dielectric layer and/or the hydrophobic coating on the dielectric layer) layer) wetting properties (also discussed above). Such frequency ranges and average or peak power ranges are known in the art. See, eg, US Patent No. 6,958,132 (Chiou et al.), US Patent No. RE44,711 (Wu et al.) (originally published as US Patent No. 7,612,355), and US Patent Application Publication No. US2014/0124370 (Short et al. Human), US2015/0306598 (Khandros et al) and US2015/0306599 (Khandros et al).growth roomNon-limiting examples of

generic growth chambers

244, 246, and 248 are shown within the microfluidic device 240 depicted in Figures 2C and 2D. Each

growth chamber

244 , 246 , and 248 may include an isolation structure 250 that defines an isolation region 258 and fluidly connects the isolation region 258 to a connection region 254 of a channel 122 . The connection region 254 may include a proximal opening 252 to the channel 122 and a distal opening 256 to the isolation region 258 . The connecting region 254 can be configured such that the maximum penetration depth for a flow from the channel 122 to one of the fluid media (not shown) in the

growth chambers

244 , 246 , 248 does not extend into the isolation region 258 . Thus, due to the connection region 254, a tiny object (not shown) or other material (not shown) disposed in an isolation region 258 of a

growth chamber

244, 246, 248 may thus interact with the channel 122. A flow of the medium 180 in the channel 122 is isolated and substantially unaffected by a flow of the medium 180 in the channel 122 . Thus, channel 122 may be an example of a swept area, and isolation area 258 of

growth chambers

244, 246, 248 may be an example of an unswept area. As mentioned herein, the channel 122 and

growth chambers

244 , 246 , 248 may be configured to contain one or more fluid media 180 . In the example shown in FIGS. 2C-2D , port 242 is connected to channel 122 and allows a fluidic medium 180 to be introduced into or removed from microfluidic device 240 . The microfluidic device may be pre-filled with a gas such as carbon dioxide gas prior to introduction of the fluid medium 180. Once the microfluidic device 240 contains the fluid medium 180, a flow 260 of the fluid medium 180 in the channel 122 can be selectively generated and stopped. For example, as shown in the figures, ports 242 can be positioned at different locations (eg, opposite ends) of the channel 122, and a flow of culture medium can be generated from one port 242 serving as an inlet to the other port 242 serving as an outlet 260. Figure 2E shows a detailed view of an example of a growth chamber 244 in accordance with the present invention. An example of a tiny object 270 is also shown in the figure. As is well known, a flow 260 of fluid medium 180 in a microfluidic channel 122 through a proximal opening 252 of the growth chamber 244 may cause a secondary flow 262 of the medium 180 to enter and/or exit the growth chamber 244 . In order to isolate tiny objects 270 in the isolation region 258 of a growth chamber 244 from the secondary flow 262, the length L of the connecting region 254 of the growth chamber 244 _con(ie, from the proximal opening 252 to the distal opening 256 ) should be greater than the penetration depth D of the secondary flow 262 into the connecting region 254 _p. Penetration depth D of substream 262 _pIt depends on the velocity of the fluid medium 180 flowing in the channel 122 and various parameters related to the configuration of the channel 122 and the proximal opening 252 of the connection region 254 to the channel 122 . For a given microfluidic device, the configuration of channel 122 and opening 252 will be fixed, while the rate of flow 260 of fluid medium 180 in channel 122 will be variable. Accordingly, for each growth chamber 244, a maximum velocity Vmax of the flow 260 of the fluid medium 180 in the channel 122 can be identified, which ensures the penetration depth D of the secondary flow 262 _pdoes not exceed the length L of the connection area 254 _con. As long as the velocity of the flow 260 of fluid medium 180 in the channel 122 does not exceed the maximum velocity Vmax, the resulting secondary flow 262 can be confined to the channel 122 and the connecting region 254 and remain out of the isolation region 258. Therefore, the flow 260 of the medium 180 in the channel 122 does not carry out the tiny objects 270 from the isolation area 258 . Specifically, the tiny objects 270 positioned in the isolation area 258 will remain in the isolation area 258 regardless of the flow 260 of the fluid medium 180 in the channel 122 . Furthermore, the flow 260 of the fluid medium 180 in the channel 122 does not move contaminant particles (eg, microparticles and/or nanoparticles) from the channel 122 to a in isolation region 258 of growth chamber 244 . Therefore, let the length L of the connection region 254 be _conGreater than the maximum penetration depth D of the secondary stream 262 _pContamination of a growth chamber 244 may be prevented by contamination of contamination by contaminant particles from channel 122 or another growth chamber (eg,

growth chambers

246, 248 in Figure 2D). Because the channel 122 and the connection area 254 of the

growth chambers

244 , 246 , 248 are affected by the flow 260 of the medium 180 in the channel 122 , the channel 122 and the connection area 254 can be viewed as the sweep (or flow) of the microfluidic device 240 area. On the other hand, the isolated regions 258 of the

growth chambers

244, 246, 248 can be considered as unswept (or no-flow) regions. For example, components in a first fluid medium 180 in channel 122 (not shown) may diffuse from channel 122 through connecting region 254 and into isolation region 258 substantially only by the components of first medium 180 A second fluid medium 280 is mixed with the second fluid medium 280 in the isolation region 258 . Similarly, components of the second medium 280 in the isolation region 258 (not shown) can diffuse from the isolation region 258 through the connecting region 254 and into the second medium in the channel 122 substantially only by the components of the second medium 280 . A medium 180 is mixed with the first medium 180 in the channel 122 . The first medium 180 may be one of the same or different mediums than the second medium 280 . Also, the first medium 180 and the second medium 280 may be the same initially and then become different (eg, by conditioning the second medium 280 by one or more cells in the isolation region 258 , or by changing the medium flowing through the channel 122 ) 180). Maximum penetration depth D of secondary flow 262 due to flow 260 of fluid medium 180 in channel 122 _pMay depend on several parameters as mentioned above. Examples of such parameters include the shape of the channel 122 (eg, the channel may direct the medium into the connection region 254 , divert the medium away from the connection region 254 , or open at the proximal end of the connection region 254 substantially perpendicular to the channel 122 ) 252 to guide the culture medium in one direction), a width W of the channel 122 at the proximal opening 252 _ch(or cross-sectional area), a width W of the connecting region 254 at the proximal opening 252 _con(or cross-sectional area), velocity V of flow 260 of fluid medium 180 in channel 122, viscosity of first medium 180 and/or second medium 280, or the like. In some embodiments, the dimensions of channel 122 and

growth chambers

244, 246, 248 may be oriented relative to the vector of flow 260 of fluid medium 180 in channel 122 as follows: channel width W _ch(or the cross-sectional area of the channel 122) may be substantially perpendicular to the flow 260 of the medium 180; the width W of the connecting region 254 at the opening 252 _con(or cross-sectional area) may be substantially parallel to the flow 260 of the medium 180 in the channel 122; and/or the length L of the connecting region _conThe flow 260 of the medium 180 in the channel 122 may be substantially perpendicular. The above orientations are examples only, and the relative positions of the channel 122 and the

growth chambers

244, 246, 248 may be in other orientations relative to each other. As shown in FIG. 2E, the width W of the connecting region 254 from the proximal opening 252 to the distal opening 256 _concan be uniform. Therefore, the width W of the connecting region 254 at the distal opening 256 _conThe width W of the connection region 254 at the proximal opening 252 may be referred to herein _conany of the identified ranges. Alternatively, the width W of the connecting region 254 at the distal opening 256 _conmay be greater than the width W of the connecting region 254 at the proximal opening 252 _con. As shown in FIG. 2E , the width W of the isolation region 258 at the distal opening 256 may be substantially the same as the width W of the connecting region 254 at the proximal opening 252 _con. Thus, the width of the isolation region 258 at the distal opening 256 may be referred to herein as the width W of the connecting region 254 at the proximal opening 252 _conany of the identified ranges. Alternatively, the width W of the isolation region 258 at the distal opening 256 may be greater or less than the width W of the connecting region 254 at the proximal opening 252 _con. Also, the distal opening 256 may be smaller than the proximal opening 252 and the width W of the connecting region 254 _conThere may be a narrowing between the proximal opening 252 and the distal opening 256 . For example, a variety of different geometries can be used to narrow the connection region 254 between the proximal and distal openings (eg, beveled connection region, beveled connection region). Further, any portion or sub-portion of the connecting region 254 can be narrowed (eg, a portion of the connecting region adjacent to the proximal opening 252). FIGS. 4A-4C depict another exemplary embodiment of a microfluidic device 400 including a microfluidic circuit 432 and flow channel 434, which are the respective microfluidic devices of FIG. 1 . Variations of device 100 , lines 132 and channels 134 . The microfluidic device 400 also has a plurality of growth chambers 436, which are additional variations of the

growth chambers

124, 126, 128, 130, 244, 246 or 248 described above. In particular, it should be understood that growth chamber 436 of device 400 shown in FIGS. 4A-4C can replace

growth chambers

124 , 126 , 128 , 130 , 244 described above in

devices

100 , 200 , 240 , and 290 , 246 or 248. Likewise, microfluidic device 400 is another variation of microfluidic device 100 and may also have the same or different DEP configurations as

microfluidic devices

100, 200, 240, 290 described above and described herein any of the other microfluidic system components. The microfluidic device 400 of FIGS. 4A-4C includes a support structure (not visible in FIGS. 4A-4C, but may be the same as or substantially similar to the support structure 104 of the device 100 depicted in FIG. 1), a microfluidic The wiring structure 412 and a cover (not visible in FIGS. 4A-4C, but may be the same as or substantially similar to the cover 110 of the device 100 depicted in FIG. 1). Microfluidic circuit structure 412 includes a frame 414 and microfluidic circuit material 416, which may be the same as or substantially similar to frame 114 and microfluidic circuit material 116 of device 100 shown in FIG. As shown in FIG. 4A , the microfluidic circuit 432 defined by the microfluidic circuit material 416 can include a plurality of channels 434 (two are shown, but more may be present) to which the plurality of growth chambers 436 are fluidly connected. . Each growth chamber 436 may include an isolation structure 446 , an isolation region 444 within the isolation structure 446 , and a connection region 442 . Connection region 442 fluidly connects channel 434 to isolation region 444 from a proximal opening 472 at channel 434 to a distal opening 474 at isolation structure 436 . In general, and in accordance with the above discussion of FIGS. 2D and 2E , a flow 482 of a first fluid medium 402 in a channel 434 may result from the channel 434 entering the respective connecting regions 442 of the growth chamber 436 and/or exiting the growth chamber 436 . The secondary streams 484 of the first medium 402 of the respective connecting regions 442 . As shown in FIG. 4B , the connection region 442 of each growth chamber 436 generally includes the region extending between a proximal opening 472 to a channel 434 and a distal opening 474 to an isolation structure 446 . The length L of the connection area 442 _conCan be greater than the maximum penetration depth D of the secondary flow 484 _p, in this case, the secondary flow 484 would extend into the connecting region 442 and not be diverted to the isolation region 444 (as shown in FIG. 4A ). Alternatively, as depicted in Figure 4C, the connecting region 442 may have a depth D less than the maximum penetration _pa length L _con, in which case the secondary flow 484 would extend through the connecting region 442 and be diverted to the isolation region 444 . In this latter case, the length L of the connecting region 442 _c1and L _c2The sum is greater than the maximum penetration depth D _p, so that the secondary flow 484 does not extend into the isolation region 444 . Regardless of the length L of the connection area 442 _congreater than the penetration depth D _por the length L of the connecting region 442 _c1and L _c2The sum is greater than the penetration depth D _p, a stream 482 of a first medium 402 in a channel 434 (which does not exceed a maximum velocity V _max) will produce a penetration depth D _pa secondary flow, and tiny objects in isolation regions 444 of a growth chamber 436 (not shown, but may be the same or substantially similar to tiny objects 270 shown in FIG. A stream 482 of the first medium 402 is carried out of the isolation region 444 . The flow 482 in the channel 434 also does not carry contaminant material (not shown) from the channel 434 into the isolation region 444 of a growth chamber 436 . Thus, the only mechanism by which components in a first medium 402 in a diffusion system channel 434 can move from channel 434 to a second medium 404 in an isolated region 444 of a growth chamber 436 . Likewise, diffusion is the only mechanism by which components in a second medium 404 in an isolated region 444 of the growth chamber 436 can move from the isolated region 444 to a first medium 402 in a channel 434 . The first medium 402 may be the same medium as the second medium 404 , or the first medium 402 may be a different medium than the second medium 404 . Alternatively, the first medium 402 and the second medium 404 may be the same initially and then become different, for example, by conditioning the second medium by one or more cells in the isolation region 444, or by changing the medium flowing through the channel 434 . As shown in FIG. 4B, the width W of the channels 434 of the channels 434 _ch(ie, taken transverse to the direction of fluid medium flowing through a channel (indicated by arrow 482 in FIG. 4A )) may be substantially perpendicular to a width W of the proximal opening 472 _con1and thus substantially parallel to a width W of the distal opening 474 _con2. However, the width W of the proximal opening 472 _con1and the width W of the distal opening 474 _con2need not be substantially perpendicular to each other. For example, the width W of the proximal opening 472 _con1An axis (not shown) oriented thereon and the width W of the distal opening 474 _con2An angle between the other axis oriented thereon may not be a right angle and thus not 90°. Examples of alternate orientation angles include angles within any of the following ranges: from about 30° to about 90°, from about 45° to about 90°, from about 60° to about 90°, or the like. In various embodiments of the growth chamber (eg, 124, 126, 128, 130, 244, 246, 248, or 436), the isolation region (eg, 258 or 444) is configured to contain a plurality of tiny objects. In other embodiments, the isolation region may be configured to contain only 1, 2, 3, 4, 5, or a similar relatively small number of tiny objects. Accordingly, the volume of an isolated region may be, for example, at least 3×10 ³Cubic microns, 6×10 ³Cubic microns, 9×10 ³Cubic micron, 1×10 ⁴Cubic microns, 2×10 ⁴Cubic microns, 4×10 ⁴Cubic microns, 8×10 ⁴Cubic micron, 1×10 ⁵Cubic microns, 2×10 ⁵Cubic microns, 4×10 ⁵Cubic microns, 8×10 ⁵Cubic micron, 1×10 ⁶Cubic microns, 2×10 ⁶Cubic microns, 4×10 ⁶Cubic microns, 6×10 ⁶Cubic micron, 1×10 ⁷Cubic microns, 2×10 ⁷Cubic microns, 4×10 ⁷Cubic microns, 6×10 ⁷Cubic micron, 1×10 ⁸cubic microns or larger. In various embodiments of the growth chamber, the width W of the

channels

122, 434 at a proximal opening (eg, 252, 472) _chMay be in any of the following ranges: 50 microns to 1000 microns, 50 microns to 500 microns, 50 microns to 400 microns, 50 microns to 300 microns, 50 microns to 250 microns, 50 microns to 200 microns, 50 microns to 150 microns , 50 microns to 100 microns, 70 microns to 500 microns, 70 microns to 400 microns, 70 microns to 300 microns, 70 microns to 250 microns, 70 microns to 200 microns, 70 microns to 150 microns, 90 microns to 400 microns, 90 microns microns to 300 microns, 90 microns to 250 microns, 90 microns to 200 microns, 90 microns to 150 microns, 100 microns to 300 microns, 100 microns to 250 microns, 100 microns to 200 microns, 100 microns to 150 microns and 100 microns to 120 microns. The above ranges are examples only, and the widths W of the

channels

122, 434 are _chOther ranges, such as one defined by any of the endpoints listed above, may be within. Moreover, the W of

channels

122, 434 _chCan be selected to be within any of these regions of the channel except at a proximal opening of a growth chamber. In some embodiments, a growth chamber has a cross-sectional height of about 30 microns to about 200 microns or about 50 microns to about 150 microns. In some embodiments, the growth chamber has a cross-sectional area of from about 100,000 microns square to about 2,500,000 microns square or from about 200,000 microns square to about 2,000,000 microns square. In some embodiments, a connecting region has a cross-sectional height that matches the cross-sectional height of the corresponding growth chamber. In some embodiments, the connection region has a cross-sectional width of about 50 microns to about 500 microns or about 100 microns to about 300 microns. In various embodiments of the growth chamber, the height H of the

channels

122, 434 at a

proximal opening

252, 472 _chMay be in any of the following ranges: 20-100 microns, 20-90 microns, 20-80 microns, 20-70 microns, 20-60 microns, 20-50 microns, 30-100 microns , 30 microns to 90 microns, 30 microns to 80 microns, 30 microns to 70 microns, 30 microns to 60 microns, 30 microns to 50 microns, 40 microns to 100 microns, 40 microns to 90 microns, 40 microns to 80 microns, 40 microns microns to 70 microns, 40 microns to 60 microns, or 40 microns to 50 microns. The above ranges are examples only, and the height H of the

channels

122, 434 _chOther ranges, such as one defined by any of the endpoints listed above, may be within. Height H of

channels

122, 434 _chCan be selected to be within any of these ranges in the region of the channel other than at a proximal opening of a growth chamber. In various embodiments of the growth chamber, a cross-sectional area of the

channels

122, 434 at a

proximal opening

252, 472 may be in any of the following ranges: 500 to 50,000 square microns, 500 to 40,000 square microns microns, 500 microns to 30,000 microns, 500 microns to 25,000 microns, 500 microns to 20,000 microns, 500 microns to 15,000 microns, 500 microns to 10,000 microns, 500 microns to 7,500 microns, 500 microns to 5,000 microns, 1,000 to 25,000 microns, 1,000 to 20,000 microns, 1,000 to 15,000 microns, 1,000 to 10,000 microns, 1,000 to 7,500 microns, 1,000 microns microns to 5,000 microns square, 2,000 microns square to 20,000 microns square, 2,000 microns square to 15,000 microns square, 2,000 microns square to 10,000 microns square, 2,000 microns square to 7,500 microns square, 2,000 microns square to 6,000 microns square, 3,000 microns square to 20,000 microns square, 3,000 microns square to 15,000 microns square, 3,000 microns square to 10,000 microns square, 3,000 microns square to 7,500 microns square or 3,000 microns square to 6,000 microns square. The above ranges are examples only, and the cross-sectional area of the channel 122 at a

proximal opening

252, 472 may be in other ranges, such as a range defined by any of the endpoints listed above. In various embodiments of the growth chamber, the length L of the connecting

regions

254, 442 _conMay be in any of the following ranges: 1 to 200 microns, 5 to 150 microns, 10 to 100 microns, 15 to 80 microns, 20 to 60 microns, 20 to 500 microns, 40 to 400 microns , 60 microns to 300 microns, 80 microns to 200 microns and 100 microns to 150 microns. The above ranges are only examples, and the length L of a connecting

region

254, 442 _conCan be within a range other than the examples above (eg, a range defined by any of the endpoints listed above). In various embodiments of the growth chamber, the width W of a connecting

region

254, 442 at a proximal opening 252 _conMay be in any of the following ranges: 20-500 microns, 20-400 microns, 20-300 microns, 20-200 microns, 20-150 microns, 20-100 microns, 20-80 microns , 20 microns to 60 microns, 30 microns to 400 microns, 30 microns to 300 microns, 30 microns to 200 microns, 30 microns to 150 microns, 30 microns to 100 microns, 30 microns to 80 microns, 30 microns to 60 microns, 40 microns microns to 300 microns, 40 microns to 200 microns, 40 microns to 150 microns, 40 microns to 100 microns, 40 microns to 80 microns, 40 microns to 60 microns, 50 microns to 250 microns, 50 microns to 200 microns, 50 microns to 150 microns, 50 microns to 100 microns, 50 microns to 80 microns, 60 microns to 200 microns, 60 microns to 150 microns, 60 microns to 100 microns, 60 microns to 80 microns, 70 microns to 150 microns, 70 microns to 100 microns and 80 microns to 100 microns. The above ranges are only examples, and the width W of a connecting

region

254, 442 at a proximal opening 252 _conCan vary from the examples above (eg, a range is defined by any of the endpoints listed above). In various embodiments of the growth chamber, the width W of a connecting

region

254, 442 at a

proximal opening

252, 472 _conMay be in any of the following ranges: 2-35 microns, 2-25 microns, 2-20 microns, 2-15 microns, 2-10 microns, 2-7 microns, 2-5 microns , 2 microns to 3 microns, 3 microns to 25 microns, 3 microns to 20 microns, 3 microns to 15 microns, 3 microns to 10 microns, 3 microns to 7 microns, 3 microns to 5 microns, 3 microns to 4 microns, 4 microns to 20 microns, 4 microns to 15 microns, 4 microns to 10 microns, 4 microns to 7 microns, 4 microns to 5 microns, 5 microns to 15 microns, 5 microns to 10 microns, 5 microns to 7 microns, 6 microns to 15 microns, 6 microns to 10 microns, 6 microns to 7 microns, 7 microns to 15 microns, 7 microns to 10 microns, 8 microns to 15 microns and 8 microns to 10 microns. The above ranges are examples only, and the width W of a connecting

region

254, 442 at a

proximal opening

252, 472 _conCan vary from the examples above (eg, a range is defined by any of the endpoints listed above). In various embodiments of the growth chamber, the length L of a connecting

region

254, 442 at the

proximal opening

252, 472 _conWidth W with one of the connecting regions 254, 442 _conA ratio may be greater than or equal to any of the following ratios: 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 6.0, 7.0, 8.0, 9.0, 10.0, or greater. The above ratios are examples only, and the length L of one of the connecting regions 254 at the

proximal openings

252, 472 _conWidth W with one of the connecting

regions

254, 442 _conThe ratio can be different from the above examples. In various embodiments of

microfluidic devices

100, 200, 240, 290, 400, V _maxcan be set to about 0.2 microliters/sec, about 0.3 microliters/sec, about 0.4 microliters/sec, about 0.5 microliters/sec, about 0.6 microliters/sec, about 0.7 microliters/sec, about 0.8 microliters /sec, about 0.9 microliters/sec, about 1.0 microliters/sec, about 1.1 microliters/sec, about 1.2 microliters/sec, about 1.3 microliters/sec, about 1.4 microliters/sec, or about 1.5 microliters/sec second. In some other embodiments, instead, V _maxcan be set to about 0.2 microliters/sec, about 0.3 microliters/sec, about 0.4 microliters/sec, about 0.5 microliters/sec, about 0.6 microliters/sec, about 0.7 microliters/sec, about 0.8 microliters /sec, about 0.9 microliters/sec, about 1.0 microliters/sec, about 1.1 microliters/sec, about 1.2 microliters/sec, about 1.3 microliters/sec, about 1.4 microliters/sec, about 1.5 microliters/sec sec, about 1.6 µl/sec, about 1.7 µl/sec, about 1.8 µl/sec, about 1.9 µl/sec, about 2.0 µl/sec, about 2.1 µl/sec, about 2.2 µl/sec , about 2.3 microliters/second, about 2.4 microliters/second, or about 2.5 microliters/second. In other embodiments, V _maxCan be set or approximately set to 2.0µl/sec, 2.2µl/sec, 2.4µl/sec, 2.6µl/sec, 2.8µl/sec, 3.0µl/sec, 3.2µl/sec, 3.4 µl/sec, 3.6 µl/sec, 3.8 µl/sec, 4.0 µl/sec, 4.2 µl/sec, 4.4 µl/sec, 4.6 µl/sec, 4.8 µl/sec, 5.0 µl L/sec, 6.0 µl/sec, 7.0 µl/sec, 8.0 µl/sec or 9.0 µl/sec. In various embodiments of microfluidic devices with growth chambers, the volume of an

isolation region

258, 444 of a growth chamber may be, for example, at least 3 x 10 ³Cubic microns, 6×10 ³Cubic microns, 9×10 ³Cubic micron, 1×10 ⁴Cubic microns, 2×10 ⁴Cubic microns, 4×10 ⁴Cubic microns, 8×10 ⁴Cubic micron, 1×10 ⁵Cubic microns, 2×10 ⁵Cubic microns, 4×10 ⁵Cubic microns, 8×10 ⁵Cubic micron, 1×10 ⁶Cubic microns, 2×10 ⁶Cubic microns, 4×10 ⁶Cubic microns, 6×10 ⁶cubic microns or larger. In various embodiments of microfluidic devices with growth chambers, the volume of a growth chamber may be about 5 x 10 ³Cubic microns, about 7×10 ³Cubic micron, about 1×10 ⁴Cubic microns, about 3×10 ⁴Cubic microns, about 5×10 ⁴Cubic microns, about 8×10 ⁴Cubic micron, about 1×10 ⁵Cubic microns, about 2×10 ⁵Cubic microns, about 4×10 ⁵Cubic microns, about 6×10 ⁵Cubic microns, about 8×10 ⁵Cubic micron, about 1×10 ⁶Cubic microns, about 2×10 ⁶Cubic microns, about 4×10 ⁶Cubic microns, about 8×10 ⁶Cubic micron, about 1×10 ⁷Cubic microns, about 3×10 ⁷Cubic microns, about 5×10 ⁷cubic microns or about 8 x 10 ⁷cubic microns or larger. In some embodiments, the microfluidic device has no more than 1 x 10 ²A growth chamber for biological cells, and the volume of a growth chamber may not exceed 2 × 10 ⁶cubic microns. In some embodiments, the microfluidic device has no more than 1 x 10 ²A growth chamber for biological cells, and a growth chamber can be no more than 4 × 10 ⁵cubic microns. In other embodiments, the microfluidic device has a growth chamber in which no more than 50 biological cells can be maintained, a growth chamber can be no more than 4 x 10 ⁵cubic microns. In various embodiments, the microfluidic device has growth chambers configured as any of the embodiments discussed herein, wherein the microfluidic device has about 100 to about 500 growth chambers, about 200 to about 1000 growth chambers Growth chambers, about 500 to about 1500 growth chambers, about 1000 to about 2000 growth chambers, or about 1000 to about 3500 growth chambers. In some other embodiments, the microfluidic device has growth chambers configured as any of the embodiments discussed herein, wherein the microfluidic device has about 1500 to about 3000 growth chambers, about 2000 to about 3500 growth chambers growth chambers, about 2500 to about 4000 growth chambers, about 3000 to about 4500 growth chambers, about 3500 to about 5000 growth chambers, about 4000 to about 5500 growth chambers, about 4500 to about 6000 growth chambers growth chambers, about 5000 to about 6500 growth chambers, about 5500 to about 7000 growth chambers, about 6000 to about 7500 growth chambers, about 6500 to about 8000 growth chambers, about 7000 to about 8500 growth chambers growth chambers, about 7500 to about 9000 growth chambers, about 8000 to about 9500 growth chambers, about 8500 to about 10,000 growth chambers, about 9000 to about 10,500 growth chambers, about 9500 to about 11,000 growth chambers growth chambers, about 10,000 to about 11,500 growth chambers, about 10,500 to about 12,000 growth chambers, about 11,000 to about 12,500 growth chambers, about 11,500 to about 13,000 growth chambers, about 12,000 to about 13,500 growth chambers, about 12,500 to about 14,000 growth chambers, about 13,000 to about 14,500 growth chambers, about 13,500 to about 15,000 growth chambers, about 14,000 to about 15,500 growth chambers, about 14,500 to about 16,000 growth chambers, about 15,000 to about 16,500 growth chambers, about 15,500 to about 17,000 growth chambers, about 16,000 to about 17,500 growth chambers, about 16,500 to about 18,000 growth chambers, about 17,000 to about 18,500 growth chambers, about 17,500 to about 19,000 growth chambers, about 18,000 to about 19,500 growth chambers, about 18,500 to about 20,000 growth chambers, about 19,000 to about 20,500 growth chambers, about 19,500 to about 21,000 growth chambers or from about 20,000 to about 21,500 growth chambers. FIG. 2F illustrates a microfluidic device 290 according to an embodiment. The microfluidic device 290 shown in FIG. 2F is a typical diagram of a microfluidic device 100 . In practice, the microfluidic device 290 and its constituent wiring elements, such as the channel 122 and the growth chamber 128, will have the dimensions discussed herein. The microfluidic circuit 120 depicted in FIG. 2F has two ports 107 , four different channels 122 , and four different flow paths 106 . The microfluidic device 290 further includes a plurality of growth chambers open to each channel 122 . In the microfluidic device depicted in Figure 2F, the growth chamber has a geometry similar to the fence depicted in Figure 2E, and thus has both connection and isolation regions. Accordingly, the microfluidic circuit 120 includes a sweep area (eg, the maximum penetration depth D of the channel 122 and the secondary flow 262 ) _ppart of connecting region 254 within) and unswept regions (eg isolation region 258 and not at maximum penetration depth D of secondary stream 262) _ppart of the connecting region 254 within) both. 3A and 3B show various embodiments of a system 150 that can be used to operate and observe microfluidic devices (eg, 100, 200, 240, 290) in accordance with the present invention. As shown in FIG. 3A, system 150 may include a structure (“nested”) 300 configured to hold a microfluidic device 100 (not shown) or any other microfluidic device described herein. Nest 300 may include a socket 302 capable of interfacing with microfluidic device 360 (eg, an optically actuated electrical device 100 ) and providing electrical connection from power source 192 to microfluidic device 360 . Nest 300 may further include an integrated electrical signal generation subsystem 304 . The electrical signal generation subsystem 304 can be configured to supply a bias voltage to the socket 302 such that when the microfluidic device 360 is held by the socket 302, the bias voltage is applied across a pair of electrodes in the microfluidic device 360 . Accordingly, the electrical signal generation subsystem 304 may be part of the power supply 192 . The ability to apply a bias voltage to the microfluidic device 360 does not mean that a bias voltage will always be applied when the microfluidic device 360 is held by the socket 302 . Rather, in most cases, the bias voltage will be applied intermittently, eg, only when needed to facilitate the generation of electrodynamic forces in the microfluidic device 360, such as dielectrophoresis or electrowetting. As shown in FIG. 3A , nest 300 may include a printed circuit board assembly (PCBA) 320 . The electrical signal generation subsystem 304 may be mounted on and electrically integrated into the PCBA 320 . The exemplary nest 300 also includes sockets 302 mounted on the PCBA 320 . Typically, the electrical signal generation subsystem 304 will include a waveform generator (not shown). The electrical signal generation subsystem 304 may further include an oscilloscope (not shown) and/or a waveform amplification circuit (not shown) configured to amplify a waveform received from the waveform generator. The oscilloscope, if present, can be configured to measure the waveform supplied to the microfluidic device 360 held by the socket 302 . In certain embodiments, the oscilloscope measures the waveform at a location close to the microfluidic device 360 (and remote from the waveform generator) to thus ensure greater accuracy in measuring the waveform actually applied to the device . For example, data obtained from the oscilloscope measurements can be provided as feedback to the waveform generator, and the waveform generator can be configured to adjust its output based on this feedback. An example of a suitable combination of waveform generator and oscilloscope is the Red Pitaya™. In some embodiments, nest 300 further includes a controller 308 , such as a microprocessor for sensing and/or controlling electrical signal generation subsystem 304 . Examples of suitable microprocessors include Arduino™ microprocessors, such as Arduino Nano™. Controller 308 may be used to perform functions and analysis or may communicate with an external host controller 154 (as shown in FIG. 1 ) to perform functions and analysis. In the embodiment shown in FIG. 3A, the controller 308 communicates with a master controller 154 through an interface 310 (eg, a plug or connector). In some embodiments, nest 300 can include an electrical signal generation subsystem 304 that includes a Red Pitaya™ waveform generator/oscilloscope unit ("Red Pitaya unit") and a waveform amplification circuit that amplifies the The Red Pitaya unit generates the waveform and transmits the amplified voltage to the microfluidic device 100 . In some embodiments, the Red Pitaya unit is configured to measure the amplified voltage at the microfluidic device 360 and then adjust its own output voltage as needed so that the measured voltage at the microfluidic device 360 is the desired value. In some embodiments, the waveform amplification circuit may have a +6.5 volt to -6.5 volt power supply generated by a pair of DC-DC converters mounted on the PCBA 320 to result in up to 13 Vpp at the microfluidic device 100 one of the signals. As shown in FIG. 3A , nest 300 may further include a thermal control subsystem 306 . Thermal control subsystem 306 can be configured to regulate the temperature of microfluidic device 360 held by nest 300 . For example, thermal control subsystem 306 may include a Peltier thermoelectric device (not shown) and a cooling unit (not shown). The Peltier thermoelectric device may have a first surface configured to interface with at least one surface of the microfluidic device 360 . The cooling unit can be, for example, a cooling block (not shown), such as a liquid-cooled aluminum block. A second surface of the Peltier thermoelectric device (eg, opposite a surface of the first surface) can be configured to interface with a surface of the cooling block. The cooling block may be connected to a fluid path 330 configured to circulate cooling fluid through the cooling block. In the embodiment shown in FIG. 3A, the nest 300 includes an inlet 332 and an outlet 334 to receive cooling fluid from an external reservoir (not shown), which is introduced into the fluid path 330 and passed through The cooling block, and then the cooling fluid is returned to the external reservoir. In some embodiments, the Peltier thermoelectric device, the cooling unit and/or the fluid path 330 may be mounted on a housing 340 of the nest 300 . In some embodiments, thermal control subsystem 306 is configured to regulate the temperature of the Peltier thermoelectric device in order to achieve a target temperature of microfluidic device 360 . For example, temperature regulation of the Peltier thermoelectric device can be achieved by a thermoelectric power supply such as a Pololu™ Thermoelectric Power Supply (Pololu Robotics and Electronics, Inc.). Thermal control subsystem 306 may include a feedback circuit, such as a temperature value provided by an analog circuit. Alternatively, the feedback circuit may be provided by a digital circuit. In some embodiments, nest 300 may include a thermal control subsystem 306 having a feedback circuit, which is a class-to-ratio voltage divider circuit (not shown), including a resistor (eg, with a 1k Ohms +/- 0.1% resistance, +/- 0.02 ppm/C0 temperature coefficient) and an NTC thermistor (eg, with 1 kOhm +/- 0.01% nominal resistance). In some examples, thermal control subsystem 306 measures voltage from the feedback circuit and then uses the calculated temperature value as input to an on-board PID control loop algorithm. The output from the PID control loop algorithm can drive, for example, both a direction pin and a PWM signal pin on a Pololu™ motor driver (not shown) to actuate the thermoelectric power supply, Thereby, the Peltier thermoelectric device is controlled. Nest 300 may include a serial port 350 that allows the microprocessor of controller 308 to communicate with an external host controller 154 via interface 310. Additionally, the microprocessor of controller 308 may communicate with electrical signal generation subsystem 304 and thermal control subsystem 306 (eg, via a Plink tool (not shown)). Thus, through the combination of controller 308 , interface 310 and serial port 350 , electrical signal generation subsystem 304 and thermal control subsystem 306 can communicate with external host controller 154 . In this manner, the main controller 154 can assist the electrical signal generation subsystem 304, among other things, by performing scaling calculations for output voltage adjustments. A Graphical User Interface (GUI) (not shown) provided via a display device 170 coupled to the external host controller 154 may be configured to draw from the thermal control subsystem 306 and the electrical signal generation subsystem 304, respectively Obtained temperature and waveform data. Alternatively or additionally, the GUI may allow the controller 308, thermal control subsystem 306, and electrical signal generation subsystem 304 to be updated. As discussed above, system 150 may include an imaging device 194 . In some embodiments, imaging device 194 includes a light modulation subsystem 422 . Light modulation subsystem 422 may include a digital mirror device (DMD) or a micro-shutter array system (MSA), either of which may be configured to receive light from a light source 420 and a photon of the received light The set is transmitted to an optical train of microscope 450. Alternatively, light modulation subsystem 422 may include a device that generates light itself (and thus does not require a light source 420), such as an organic light emitting diode display (OLED), a liquid crystal on silicon (LCOS) device, a A ferroelectric liquid crystal device (FLCOS) or a transmissive liquid crystal display (LCD). Light modulation subsystem 422 can be, for example, a projector. Thus, the light modulation subsystem 422 is capable of emitting both structured and unstructured light. An example of a suitable light modulation subsystem 422 is the Mosaic™ system from Andor Technologies™. In some embodiments, the imaging module 164 and/or the power module 162 of the system 150 can control the light modulation subsystem 422 . In some embodiments, imaging device 194 further includes a microscope 450 . In these embodiments, nest 300 and light modulation subsystem 422 may be individually configured for mounting on microscope 450 . Microscope 450 can be, for example, a standard research grade light microscope or a fluorescence microscope. Accordingly, nest 300 can be configured to be mounted on stage 426 of microscope 450 and/or light modulation subsystem 422 can be configured to be mounted on a port of microscope 450. In other embodiments, the nest 300 and light modulation subsystem 422 described herein may be integrated components of the microscope 450. In some embodiments, microscope 450 may further include one or more detectors 440 . In some embodiments, the detector 440 is controlled by the imaging module 164 . Detector 440 may include an eyepiece, a charge coupled device (CCD), a camera (eg, a digital camera), or any combination thereof. If there are at least two detectors 440, one detector can be, for example, a fast frame rate camera and the other detector can be a high sensitivity camera. Additionally, microscope 450 may include a string of optical elements configured to receive reflected and/or emitted light from microfluidic device 360 and focus at least a portion of the reflected and/or emitted light on one or more detectors device 440. The optical string of the microscope can also include different tube lenses (not shown) for the different detectors, so that the final magnification on each detector can be different. In certain embodiments, imaging device 194 is configured to use at least two light sources. For example, a first light source 420 may be used to generate structured light (eg, via light modulation subsystem 422) and a second light source 430 may be used to provide unstructured light. The first light source 420 can generate structured light for optically actuated electrodynamic and/or fluorescent excitation, and the second light source 430 can be used to provide bright field illumination. In these embodiments, the power module 164 may be used to control the first light source 420 and the imaging module 164 may be used to control the second light source 430 . The string of optical elements of microscope 450 can be configured to: (1) receive structured light from light modulation subsystem 422 and structure when a microfluidic device, such as an optically actuated electromechanical device, is held by nest 300 focusing light on at least a first region in the device; and (2) receiving reflected and/or emitted light from the microfluidic device and focusing at least a portion of the reflected and/or emitted light to detector 440 superior. The optical element string can be further configured to receive unstructured light from a second light source and focus the unstructured light on at least a second region of the device when the microfluidic device is held by the nest 300 . In some embodiments, the first region and the second region of the microfluidic device may be overlapping regions. For example, the first region may be a subset of the second region. Shown in FIG. 3B is a first light source 420 that supplies light to a light modulation subsystem 422 that provides structured light to a string of optical elements of microscope 450 of system 450 . Shown is a second light source 430 that provides non-structure to the optical string via a beam splitter 424 . The structured light from light modulation subsystem 422 and the unstructured light from second light source 430 travel together from beam splitter 424 through the string of optical elements to a second beam splitter 424 (or dichroic filter 448, depending on for the light provided by the light modulation subsystem 422 ), where the light is reflected to pass down through the objective lens 454 to the sample plane 428 . The reflected and/or emitted light from sample plane 428 then travels back up through objective lens 454 , through beam splitter and/or dichroic filter 448 to dichroic filter 452 . Only a portion of the light reaching dichroic filter 452 passes through and reaches detector 440 . In some embodiments, the second light source 430 emits blue light. Using an appropriate dichroic filter 452, blue light reflected from the sample plane 428 can pass through the dichroic filter 452 to the detector 440. In contrast, structured light from light modulation subsystem 422 is reflected from sample plane 428 but does not pass through dichroic filter 452. In this example, dichroic filter 452 filters out visible light having a wavelength longer than 495 nanometers. Filtering this light from the light modulation subsystem 422 will only be done if the light emitted from the light modulation subsystem does not contain any wavelengths shorter than 495 nanometers (as shown in the figure). In fact, if the light from light modulation subsystem 422 includes wavelengths shorter than 495 nanometers (eg, blue light wavelengths), some of the light from the light modulation subsystem will pass through filter 452 to detector 440 . In this embodiment, the filter 452 is used to change the balance between the amount of light reaching the detector 440 from the first light source 420 and the second light source 430 . This may be beneficial if the first light source 420 is significantly stronger than the second light source 430 . In other embodiments, the second light source 430 may emit red light, and the dichroic filter 452 may filter visible light other than red light (eg, visible light having a wavelength shorter than 650 nanometers). Additional system components for maintaining cell viability in growth chambers of microfluidic devicesTo promote growth and/or expansion of cell populations, environmental conditions conducive to maintaining functional cells may be provided by additional components of the system. For example, these additional components can provide nutrients, cell growth signaling species, pH modulation, gas exchange, temperature control, and removal of waste from cells. Conditioning surfaces for microfluidic devicesIn some embodiments, at least one surface of the microfluidic device is conditioned to support cell growth, viability, transplantability, or any combination thereof. In some embodiments, substantially all of the inner surface is conditioned. A conditioning surface can be one of the elements that facilitate successful cell culture within a microfluidic device. Identifying an appropriate conditioning surface requires balancing several operational requirements. First, the conditioning surface can provide a contact surface to shield cells from the types of materials that can be used to fabricate such microfluidic devices. Without being bound by theory, the conditioning surface may be surrounded by water of hydration that provides a water contact layer with the cells rather than a metal contact layer. Second, the conditioning surface can provide a contact surface that can be used to properly support at least one biological cell during incubation without substantially inhibiting the ability to remove cells from the growth chamber after incubation is complete. For example, many cells require a contact surface with a certain degree of hydrophilicity to be sufficiently attached to survive and/or grow. Instead, some cells require a contact surface with a certain degree of hydrophobicity to grow and to exhibit the desired level of viability. Additionally, some cells require the presence of selected protein or peptide motifs within the contact surface to initiate a survival/growth response. Third, conditioning at least one surface may allow the power used in the microfluidic device to function substantially within the normal operating power range. For example, if light-actuated power is employed, the conditioning surface may substantially allow light to pass through the conditioning surface such that the light-actuated power is substantially uninhibited. The at least one conditioning surface may comprise a surface of the growth chamber or a surface of the flow region, or a combination thereof. In some embodiments, each of the plurality of growth chambers has at least one conditioning surface. In other embodiments, each of the plurality of flow regions has at least one conditioning surface. In some embodiments, at least one surface of each of the plurality of growth chambers and each of the plurality of flow regions is a conditioning surface. Conditioning surface comprising a polymerAt least one conditioning surface may comprise a polymer. The polymer may be covalently or non-covalently bonded to at least one surface. Polymers can have various structural units, including block polymers (and copolymers), star polymers (star copolymers), and graft or comb polymers (graft copolymers), among others All are suitable for use herein. The polymer may comprise a polymer having an alkylene ether moiety. Various alkylene ether-containing polymers can be suitable for use in the microfluidic devices described herein. One non-limiting exemplary class of alkylene ether-containing polymers are amphiphilic nonionic block copolymers comprising polyethylene oxide (PEO) in different ratios and in different positions within the polymer chain. ) subunits and blocks of polypropylene oxide (PPO) subunits. Pluronic® polymers (BASF) are block copolymers of this type and are considered suitable in the art for use in contact with living cells. The average molecular weight of the polymer M _wIn the range from about 2000 Da to about 20 KDa. In some embodiments, the PEO-PPO block copolymer can have a hydrophilic-lipophilic balance (HLB) greater than about 10 (eg, 12 to 18). Pluronic® polymers used to create a conditioning surface include Pluronic® L44, L64, P85 and F127 (which includes F127NF). Another class of polymers containing alkylene ethers is polyethylene glycol (PEG, Mw < 100,000 Da) or alternatively polyethylene oxide (PEO, M _w>100,000). In some embodiments, a PEG can have one of about 1000 Da, about 5000 Da, about 10,000 Da, or about 20,000 Da, M _w. In other embodiments, the polymer conditioning surface may comprise a polymer containing a carboxylic acid moiety. The carboxylic acid subunit may be one of the subunits containing an alkyl moiety, an alkenyl moiety, or an aromatic moiety. A non-limiting example is polylactic acid (PLA). In some other embodiments, the polymer conditioning surface may comprise a polymer containing a urethane moiety, such as, but not limited to, a polyurethane. In other embodiments, the polymer conditioning surface may comprise a polymer containing a sulfonic acid moiety. The sulfonic acid subunit may be one of the subunits containing an alkyl moiety, an alkenyl moiety, or an aromatic moiety. A non-limiting example is polystyrene sulfonic acid (PSSA) or polyanethole sulfonic acid. These latter exemplary polymers are polyelectrolytes and can modify the properties of the surface to assist/prevent adhesion. In other embodiments, the polymer conditioning surface may comprise a polymer containing phosphoric acid moieties at one end of the polymer backbone or at a side chain from the polymer backbone. In other embodiments, the polymer conditioning surface may comprise a polymer containing a carbohydrate moiety. In a non-limiting example, polysaccharides, such as those derived from algae or fungi, such as xanthan gum or polydextrose, may be suitable for use in forming a polymer modulating surface that may aid or prevent cell adhesion. For example, polydextrose polymers having a size of about 3 KDa can be used to provide a conditioning surface within a microfluidic device. In other embodiments, the polymeric modulating surface may comprise a polymer containing nucleotide moieties (ie, nucleic acids), which may have ribonucleotide moieties or deoxyribonucleotide moieties. Nucleic acids may contain only natural nucleotide moieties or may contain non-natural nucleotide moieties including nucleobase, ribose or phosphate moiety analogs such as (but not limited to) 7-deazaadenine, pentose, methyl Phosphonate or phosphorothioate moieties. A polymer containing nucleic acid may contain a polyelectrolyte to assist or prevent adhesion. In other embodiments, the polymer conditioning surface may comprise a polymer containing amino acid moieties. The polymer containing amino acid moieties may comprise a polymer containing natural amino acids or a polymer containing non-natural amino acids, any of which may comprise peptides, polypeptides or proteins. In a non-limiting example, the protein can be bovine serum albumin (BSA). In some embodiments, an extracellular matrix (ECM) protein can be provided within the regulatory surface for optimal cell adhesion promoting cell growth. A cell matrix protein that can be included in a regulatory surface can include, but is not limited to, a collagen, an elastin, an RGD-containing titanium (eg, a fibronectin), or a layer of cohesin. In other embodiments, growth factors, cytokines, endocrines, or other cellular signaling species may be provided within at least one conditioning surface of the microfluidic device. In further embodiments, the polymer conditioning surface may comprise a polymer having amine moieties. The polyamine polymer may comprise a natural polyamine polymer or a synthetic polyamine polymer. Examples of natural polyamines include spermine, spermidine, and putrescine. In some embodiments, the polymer conditioning surface may comprise a polymer comprising one or more of the following: alkylene oxide moieties, carboxylic acid moieties, sulfonic acid moieties, phosphate moieties, carbohydrate moieties, nucleotide moieties, or amines base acid moiety. In other embodiments, the polymer conditioning surface may comprise a mixture of one or more polymers each having one or more of the following: alkylene oxide moieties, carboxylic acid moieties, sulfonic acid moieties, phosphate moieties, carbohydrate moieties, nucleotides Moieties and/or amino acid moieties, etc., can be incorporated into the conditioning surface independently or simultaneously. covalently linked Of conditioning surfaceIn some embodiments, at least one conditioning surface comprises a covalently linked moiety configured to support cell growth, viability, transplantability, or any combination thereof, of one or more biological cells within the microfluidic device. The covalently linking moiety may comprise a linking group, wherein the linking group is covalently linked to a surface of the microfluidic device. The linking group is also linked to the moiety that is configured to support cell growth, viability, transplantability, or any combination thereof, of one or more biological cells within the microfluidic device. The surface to which the linking group is bonded may comprise a surface of the substrate of the microfluidic device, which may comprise silicon and/or silicon dioxide for embodiments in which the microfluidic device comprises a DEP configuration. In some embodiments, the covalent linkage modulating surface comprises the entire inner surface of the microfluidic device. A schematic diagram of a microfluidic device with a conditioning surface is shown in FIG. 9 . As seen in FIG. 9, a microfluidic device 900 has a first DEP substrate 904 and a second DEP substrate 906 facing an enclosed region 902 of the microfluidic device, the microfluidic device 900 may include at least one growth chamber and/or flow area. Device 900 may be configured in other ways, as with any of

microfluidic devices

100 , 200 , 240 , 290 , 400 , 500A-500E or 600 . The enclosed area 902 may be an area in which biological cells are maintained or into or from which biological cells are imported. The inner surface 910 (of the second DEP substrate 906) and the inner surface 912 (of the first DEP substrate 904) are modified with a conditioning surface 916, which may support cell growth, viability, transplantability, or the like any part of any combination. In this embodiment, the conditioning surface is covalently bonded to the oxidized functional groups of the inner surface via siloxy linking groups 914. In some embodiments, covalently linked moieties configured to support cell growth, viability, transplantability, or any combination thereof, may comprise: alkyl or fluoroalkyl (which includes perfluoroalkyl) moieties Monosaccharides or polysaccharides (which may include, but are not limited to, polydextrose); alcohols (which include, but are not limited to, propargyl alcohol); polyols, which include, but are not limited to, polyvinyl alcohol; ethers, which include, but are not limited to, polyethylene glycol; polyelectrolytes, which include, but are not limited to, polyacrylic acid or polyvinylphosphonic acid; amine groups, which include derivatives thereof, such as, but are not limited to, alkyl groups amines, hydroxyalkylated amines, guanidine salts, and heterocyclic groups containing an unaromatic nitrogen ring atom, such as (but not limited to) morpholinyl or piperazinyl); carboxylic acids, including (but not limited to) ) propynoic acid (which can provide a carboxylate anion surface); phosphonic acid, including but not limited to ethynylphosphonic acid (which can provide a phosphonate anion surface); sulfonic acid anion; carboxybetaine; sulfobetaine ; amine sulfonic acid; or amino acid. The covalently linked moiety configured to support cell growth, viability, transplantability, or any combination thereof of one or more biological cells within the microfluidic device may be any polymer as described herein, And may comprise one or more polymers containing alkylene oxide moieties, carboxylic acid moieties, carbohydrate moieties, sulfonic acid moieties, phosphoric acid moieties, amino acid moieties, nucleic acid moieties or amine moieties. In other embodiments, covalently linked moieties configured to support cell growth, viability, transplantability, or any combination thereof of one or more biological cells may comprise non-polymeric moieties, such as alkyl moieties, Fluoroalkyl moieties (which include, but are not limited to, perfluoroalkyl groups), amino acid moieties, alcohol moieties, amine moieties, carboxylic acid moieties, phosphonic acid moieties, sulfonic acid moieties, amine sulfonic acid moieties, or carbohydrate moieties. In some embodiments, the covalently linked moiety can be an alkyl group. The alkyl group can include carbon atoms forming a straight chain (eg, at least 10 carbons or a straight chain of at least 14, 16, 18, 20, 22, or more carbons). Thus, the alkyl group can be an unbranched alkyl group. In some embodiments, the alkyl group may comprise a substituted alkyl group (eg, some of the carbons in the alkyl group may be fluorinated or perfluorinated). The alkyl group can include a straight chain of substituted (eg, fluorinated or perfluorinated) carbon bonded to a straight chain of unsubstituted carbon. For example, the alkyl group may comprise a first segment (which may comprise a perfluoroalkyl group) joined to a second segment (which may comprise an unsubstituted alkyl group). The first segment and the second segment may be joined directly or indirectly (eg, via an ether linkage). The first segment of the alkyl group can be located at the distal end of the linking group, and the second segment of the alkyl group can be located at the proximal end of the linking group. In other embodiments, the alkyl group may comprise a branched alkyl group and may further have one or more arylidene groups interrupting the alkyl backbone of the alkyl group. In some embodiments, the branched or aryl-extended moiety of the alkyl or fluorinated alkyl is positioned at a point distal to the covalent bond to the surface. In other embodiments, the covalently linked moiety may comprise at least one amino acid, which may comprise more than one amino acid. Covalently linked moieties may comprise peptides or proteins. In some embodiments, the covalently linked moiety can comprise an amino acid, which can provide a zwitterionic surface for supporting cell growth, viability, transplantability, or any combination thereof. The covalently linked moiety may contain one or more sugars. Such covalently linked sugars may be monosaccharides, disaccharides or polysaccharides. The covalently linked sugars can be modified to introduce a reactive partner moiety that allows coupling to or (in detail) attachment to the surface. Exemplary reaction partner moieties can include aldehyde, alkynyl, or halo moieties. The polysaccharide can be modified in a random manner, wherein each of the saccharide monomers can be modified or only a portion of the saccharide monomers within the polysaccharide can be modified to provide a reactive partner moiety that can be coupled directly or indirectly to a surface . An example may include polydextrose polysaccharides that can be indirectly coupled to a surface via an unbranched linker. The covalently linked moiety may contain one or more amine groups. The amine group can be a substituted amine moiety, a guanidine moiety, a nitrogen-containing heterocyclic moiety, or a heteroaryl moiety. The amine group-containing moiety may have a structure that allows for pH modification of the environment within the microfluidic device and, optionally, the growth chamber. The covalently linked moiety may comprise one or more carboxylic acid, phosphonic acid, amine sulfonic acid or sulfonic acid moieties. In some embodiments, a covalently linked moiety can comprise one or more nucleic acid moieties, which can have a sequence of individual nucleotides designed to capture nucleic acids from biological cells within a microfluidic device. The capture nucleic acids can have a nucleotide sequence that is complementary to nucleic acids from biological cells and can capture nucleic acids by hybridization. The conditioning surface may consist of only one part or may comprise more than one different part. For example, a fluoroalkyl modulating surface (which includes a perfluoroalkyl group) can have a plurality of covalently bonded moieties that are all the same, eg, have the same covalent attachment to the surface and have the same number of fluoromethylene units ( It includes fluoroalkyl moieties that support growth and/or viability and/or transplantability). Alternatively, the conditioning surface may have more than one portion attached to the surface. For example, the conditioning surface may comprise an alkyl or fluoroalkyl group having a specified number of methylene or fluoromethylene units and may further comprise another set of moieties attached to the surface having a charged moiety attached to alkyl or fluoroalkyl chains having a greater number of methylene or fluoromethylene units. In some embodiments, conditioning surfaces with more than one attachment moiety can be designed to have a larger number of backbone atoms and thus have a first set of attachment partners that self-covalently attach to a larger length of the surface Site groups can provide capacity to present the bulkier moiety at the conditioning surface, while a second set of attached ligands with different ends that require less space and have fewer backbone atoms can help make the overall substrate Surface functionalization to prevent unwanted adhesion or contact with a silicon or aluminum oxide substrate itself. In another example, the portion attached to the surface can provide a zwitterionic surface to present alternating charges on the surface in a random manner. Adjust surface propertiesIn some embodiments, the covalently bonded moiety can form a monolayer when covalently bonded to a surface of a microfluidic device (eg, a DEP-structured substrate surface). In some embodiments, the conditioning surface formed by the covalently linked moieties can have a thickness of less than 10 nanometers (eg, less than 5 nanometers or about 1.5 nanometers to about 3.0 nanometers). In other embodiments, the conditioning surface formed by the covalently linked moieties may have a thickness of about 10 nanometers to about 50 nanometers. In some embodiments, the conditioning surface does not require a perfectly formed monolayer to operate properly within a DEP configuration. In various embodiments, the conditioning surface(s) of the microfluidic device can provide the desired electrical properties. Without being bound by theory, one factor affecting the robustness of a conditioning surface is intrinsic charge trapping. Different surface modulating materials can trap electrons that would cause the material to break down. Defects in the conditioning surface can lead to charge trapping and further breakdown of the conditioning surface. In addition to adjusting the composition of the surface, other factors, such as the physical thickness of the hydrophobic material, can also affect the DEP force. Various factors can alter the physical thickness of the conditioning surface, such as the manner in which the conditioning surface is formed on the substrate (eg, vapor deposition, liquid deposition, spin coating, flooding, and electrostatic coating). An ellipsometer can be used to measure the physical thickness and uniformity of the conditioning surface. In addition to modulating the electrical properties of the surface, the modulating surface can also have properties that are beneficial for use with biomolecules. For example, a conditioning surface containing a fluorinated (or perfluorinated) carbon chain may provide one of the benefits of reducing the amount of surface fouling over alkyl terminated chains. As used herein, surface fouling refers to the amount of indiscriminate material deposited on the surface of a microfluidic device, which may include the permanence of biological materials such as proteins and their degradants, nucleic acids and their respective degradants and the like or semi-permanent deposits. Various properties of conditioning surfaces that can be used in DEP configurations are included in the table below. As can be seen in the table, for entries 1 to 7 (all of which are covalently linked modulating surfaces as described herein), the thickness measured by ellipsometry is consistently thinner than entry 8 (by non-covalent The thickness of a CYTOP surface formed by spin coating) (N/A means no data available in the entire table). We have found that fouling is more dependent on the chemistry of the surface than the pattern of formation, since fluorinated surfaces are generally less fouling than alkane (hydrocarbon) conditioned surfaces. surface 1.Properties of various conditioning surfaces prepared by covalently modifying a surface compared to a non-covalently formed CYTOP surface

Surface modification type Surface Modifier Form thickness fouling 1. Alkyl terminated siloxane (C ₁₈ ) CH ₃ -(CH ₂ ) ₁₇ -Si-(OCH ₃ ) ₃ ~ 2 nm More fouling than fluorinated layers 2. Fluoroalkyl terminated alkylsiloxanes (C ₁₆ F) CF ₃ -(CF ₂ ) ₁₃ -(CH ₂ ) ₂ -Si-(OCH ₃ ) ₃ ~2 nm More fouling resistant than alkyl terminated layers 3. Fluoroalkyl terminated alkylsiloxane C ₁₀ F CF ₃ -(CF ₂ ) ₇ -(CH ₂ ) ₂ -Si-(OCH ₃ ) ₃ ~1 nm More fouling resistant than alkyl terminated layers 4. Alkyl terminated siloxane (C ₂₂ ) CH ₃ -(CH ₂ ) ₂₁ -Si-(OCH ₂ CH ₃ ) ₃ ~2-2.5 nm More fouling than fluorinated layers 5. Alkyl terminated siloxane (C ₁₆ ) CH ₃ -(CH ₂ ) ₁₅ -Si-(OCH ₃ ) ₃ N/A More fouling than fluorinated layers 6. Fluoroalkyl terminated alkoxyalkylsiloxane C ₆ FC ₁₃ CF ₃ -(CF ₂ ) ₅ -(CH ₂ ) ₂ -O-(CH ₂ ) ₁₁ -Si(OCH ₃ ) ₃ ~2 nm N/A 7. Alkyl terminated phosphonate C ₁₈ PA CH ₃ -(CH ₂ ) ₁₇ -P=O(OH) ₂ N/A More fouling than fluorinated layers 8. CYTOP Fluoropolymers1 ^{, 2} ~30 nm More fouling resistant than alkyl terminated layers

1. CYTOP structure:

2. Spin coating, non-covalent. bonding group to the surface groupThe covalently linked moieties forming the conditioning surface are attached to the surface via a linking group. The linking group may be a siloxy linking group formed by reacting a siloxane-containing reagent with oxides of the substrate surface (which may be formed from silicon or aluminum oxide). In some other embodiments, the linking group can be a phosphonate ester formed by reacting a phosphonic acid-containing reagent with an oxide on the surface of a silicon or aluminum substrate. Multi-Part Conditioning SurfaceThis can be accomplished by a surface conditioning agent (which is configured to already contain moieties that provide a conditioning surface) such as an alkylsiloxane agent or a fluorine-substituted alkylsiloxane agent, which may include a perfluorosiloxane agent ) to form a covalently linked modulating surface, as will be described below. Alternatively, the modulating surface can be formed by coupling moieties that support cell growth, viability, transplantability, or any combination thereof, to a surface-modifying ligand (which itself is covalently bonded to the surface). A structure and preparation method of a conditioning surfaceIn some embodiments, a conditioning surface of the oxide covalently bonded to the surface of the DEP substrate has a structure of one of Formula 1:

Formula 1 The conditioning surface can be covalently bonded to the oxide of the surface of the DEP substrate. The dielectrophoretic substrate can be silicon or aluminum oxide, and the oxide can be present as part of the substrate's natural chemical structure or can be introduced as will be discussed below. The conditioning surface may be attached to the oxide via a linking group LG, which may be a siloxyl or phosphonate group formed by reacting a siloxane or phosphonate group with the oxide. Moieties that are configured to support cell growth, viability, transplantability, or any combination thereof, may include: alkyl or fluoroalkyl (which includes perfluoroalkyl) moieties; monosaccharide or polysaccharide (which may include ( but not limited to) polydextrose); alcohols (including but not limited to propargyl alcohol); polyols, including but not limited to polyvinyl alcohol; alkylene ethers, including but not limited to polyvinyl alcohol ethylene glycol; polyelectrolytes (which include, but are not limited to, polyacrylic acid or polyvinylphosphonic acid); amines (which include derivatives thereof, such as, but not limited to, alkylated amines, hydroxyalkylated amines, Guanidine salts and heterocyclic groups containing an unaromatic nitrogen ring atom such as, but not limited to, morpholinyl or piperazinyl); carboxylic acids, including but not limited to propyonic acid (which can provide carboxylic acids anionic surfaces); phosphonic acids including, but not limited to, ethynylphosphonic acids (which can provide phosphonic acid anionic surfaces); sulfonic acid anions; carboxybetaines; sulfobetaines; amine sulfonic acids; or amino acids. The alkyl or fluoroalkyl moiety can have a backbone length equal to or greater than one of 10 carbons. In some embodiments, the alkyl or fluoroalkyl moiety can have a backbone length of one of about 10, about 12, about 14, about 16, about 18, about 20, or about 22 carbons. The linking group LG can be attached directly or indirectly to a moiety that provides support for cell growth, viability, transplantability, or any combination thereof, within the microfluidic device. When the linking group LG is directly attached to the moiety, the optional linker L is absent and n is 0. When the linking group LG is indirectly attached to the moiety, the linker L is present and n is 1. The linker L may have a linear moiety, wherein a backbone of the linear moiety may comprise a main chain selected from the group consisting of silicon atoms, carbon atoms, nitrogen atoms, oxygen atoms, sulfur atoms and phosphorus atoms (subject to chemical bonding known in the art). limit) from 1 to 200 non-hydrogen atoms in any combination. In some non-limiting examples, the linker L can be interrupted using any combination of one or more moieties selected from the group consisting of ether, amine, carbonyl, amido, or phosphonate. In addition, the linker L may have one or more arylidene groups, heteroarylidene groups or heterocyclic groups that interrupt the main chain of the linker. In some embodiments, the backbone of the linker L may contain 10 to 20 atoms. In other embodiments, the backbone of the linker L may comprise from about 5 atoms to about 200 atoms, from about 10 atoms to about 80 atoms, from about 10 atoms to about 50 atoms, or from about 10 atoms to about 40 atoms. In some embodiments, the backbone atoms are all carbon atoms. In other embodiments, the backbone atoms are not all carbon, but may include any feasible combination of silicon, carbon, nitrogen, oxygen, sulfur, or phosphorus atoms subject to chemical bonding constraints known in the art . Surface Conditioning ReagentsA surface conditioning agent of Formula 6 is useful when a moiety configured to support cell growth, viability, transplantability, or any combination thereof, and thereby provide a conditioning surface, is added to the surface of a substrate in a one-step procedure to introduce conditioning surfaces. The surface-modifying agent may have one of the structures of Formula 6:

. Formula 6 In the surface conditioning agent of formula 6, the surface conditioning agent may comprise a linking group LG, which may be a siloxane or a phosphonic acid group. The linking group LG can be linked directly or indirectly to a moiety configured to support cell growth, viability, transplantability, or any combination thereof. LG can be linked directly (n=0) or indirectly (n=1) to a conformation to support cell growth, viability, transplantability, or the like by being linked to a first end of a linker L part of any combination. The linker L may further comprise a linear moiety, wherein a main chain of the linear moiety may have a main chain selected from the group consisting of silicon atoms, carbon atoms, nitrogen atoms, oxygen atoms, sulfur atoms and phosphorus atoms (subject to chemistries known in the art). 1 to 200 non-hydrogen atoms in any combination of bonding limits). The backbone of the linear moiety may further comprise one or more arylidene moieties. Moieties ("moieties") configured to support cell growth, viability, transplantability, or any combination thereof, may comprise: alkyl or fluoroalkyl (including perfluoroalkyl) moieties; monosaccharides or polysaccharides (which may include, but are not limited to, polydextrose); alcohols (which include, but are not limited to, propargyl alcohol); polyols, which include, but are not limited to, polyvinyl alcohol; alkylene ethers, which include ( but not limited to) polyethylene glycol; polyelectrolyte (which includes but is not limited to polyacrylic acid or polyvinylphosphonic acid); amine group (which includes its derivatives such as (but not limited to) alkylated amine groups, hydroxyalkanes alkylated amine groups, guanidine salts, and heterocyclic groups containing an unaromatic nitrogen ring atom, such as, but not limited to, morpholinyl or piperazinyl); carboxylic acids, including but not limited to propyonic acid ( which can provide a carboxylate anion surface); phosphonic acids, including but not limited to, ethynylphosphonic acid (which can provide a phosphonate anion surface); sulfonic acid anions; carboxybetaines; sulfobetaines; sulfamic acids; or amino acids. Moieties that are configured to support cell growth, viability, transplantability, or any combination thereof, may comprise alkyl or perfluoroalkyl moieties. The alkyl or perfluoroalkyl moieties can have a backbone length of greater than 10 carbons. The portion of the surface modulating agent that is configured to support cell growth, viability, transplantability, or any combination thereof, may comprise a carbohydrate moiety, and may be polydextrose. In other embodiments, the portion of the surface modulating agent configured to support cell growth, viability, transplantability, or any combination thereof, may comprise an alkylene ether moiety. The isoalkylene ether moiety may be polyethylene glycol. The surface modulating agent may further comprise a cleavable moiety, which may be located within the linker L or may be part of the portion of the surface modulating agent configured to support cell growth, viability, transplantability, or any combination thereof. part. The cleavable moiety can be configured to allow disruption of the regulatory surface, thereby promoting the transplantability of one or more biological cells. In some embodiments, moieties that support cell growth, viability, transplantability, or any combination thereof, can be added to the surface of the substrate in a multi-step procedure. When the moiety is coupled to the surface in a stepwise fashion, the linker L may further comprise a coupling group CG, as shown in formula 2.

Formula 2 In some embodiments, the coupling group CG represents R from a reactive moiety _xand is configured to react with a paired moiety R _pxThe resulting portion of the reaction of one part of the reaction. For example, a typical CG may contain a carboxamide group, which is the result of the reaction of the amine group with a derivative of a carboxylic acid, such as an active ester, acyl chloride, or the like. CG may comprise triazolyl, carboxamido, thioamido, oxime, alkylthio, disulfide, ether or alkenyl or any other suitable group. The coupling group CG can be positioned at the second end of the linker L of the attached moiety therein. In some other embodiments, the coupling group CG can interrupt the backbone of the linker L. In some embodiments, the coupling group CG is triazolyl, which is the result of a reaction between an alkynyl group and an azide group, either of which can be used as known in the art. A reactive moiety or reactive pairing moiety in a click-coupling reaction. The triazolyl group can also be further substituted. For example, a dibenzocyclooctenyl fused triazolyl moiety can be derived from having a dibenzocyclooctynyl reaction partner moiety R _pxOne adjusts the azide group reaction moiety R of the modifier and the surface modified ligand _xThe reaction, as will be described in more detail in the following paragraphs. Various dibenzocyclooctynyl modified molecules are known in the art or can be synthesized to incorporate a portion that is configured to support cell growth, viability, transplantability, or any combination thereof, or the like. When forming a conditioning surface in a multi-step procedure, support can be introduced by the reaction of a conditioning modifier (Equation 5) with a substrate (which has a surface-modifying ligand covalently bonded thereto) As part of cell growth, viability, transplantability, or any combination thereof, the surface-modifying ligand has a structure of one of Formula 3.

Equation 5 Equation 3 The intermediate modified surface of formula 3 has attached to one of its surface modified ligands, which has -LG-(L'') _j-R _xOne formula, which is bonded to the oxide of the substrate, and is similarly formed as described above for the conditioning surface of formula 1. The surface of the DEP substrate is as described above and contains oxides that are native to or incorporated into the substrate. The linking group LG is as described above. The one-key linkage L'' may exist (j=1) or not (j=0). The linker L'' may have a straight-chain moiety, wherein a backbone of the straight-chain moiety may comprise a main chain selected from the group consisting of silicon atoms, carbon atoms, nitrogen atoms, oxygen atoms, sulfur atoms and phosphorus atoms (subject to known in the art). 1 to 100 non-hydrogen atoms in any combination of chemical bonding limits). In some non-limiting examples, any combination of ether, amine, carbonyl, amido, or phosphonate groups can be used to interrupt the linker L''. In addition, the linker L'' may have one or more arylidene groups, heteroarylidene groups or heterocyclic groups that interrupt the main chain of the linker. In some embodiments, the backbone of the linker L'' may contain 10 to 20 atoms. In other embodiments, the backbone of the linker L″ may comprise about 5 atoms to about 100 atoms, about 10 atoms to about 80 atoms, about 10 atoms to about 50 atoms, or about 10 atoms atom to about 40 atoms. In some embodiments, the backbone atoms are all carbon atoms. In other embodiments, the backbone atoms are not all carbon, but may include any feasible combination of silicon, carbon, nitrogen, oxygen, sulfur, or phosphorus atoms subject to chemical bonding constraints known in the art . a reaction part R _xAt the end of the surface-modifying ligand present at the distal end of the covalent bond between the surface-modifying ligand and the surface. Reaction part R _xAny suitable reactive moiety for coupling reactions to introduce moieties that support cell growth, viability, transplantability, or any combination thereof. In some embodiments, the reactive moiety R _xCan be an azide, amine, bromo, thiol, active ester, succinimidyl or alkynyl moiety. adjust modifierModulators (Formula 5) are configured to supply moieties that support cell growth, viability, transplantability, or any combination thereof.

Formula 5 The moiety of the modulating modifier configured to support cell growth, viability, transplantability, or any combination thereof, is via a reaction pairing moiety R _pxwith reactive moiety R _xThe reaction will bond to the surface modified ligand. reaction pairing part R _pxis configured to react with the respective moiety R _xAny suitable reactive portion of the reaction. In a non-limiting example, a suitable reaction pairing moiety R _pxcan be alkynyl and the reactive moiety R _xCan be azide. Alternatively, the reaction pairing moiety R _pxcan be an azido moiety and each reactive moiety R _xCan be alkynyl. In other embodiments, the reaction pairing moiety R _pxCan be an active ester functional group and the reactive moiety R _xCan be an amine group. In other embodiments, the reaction pairing moiety R _pxCan be an aldehyde group and the reactive moiety R _xCan be an amine group. Other reactive moiety-reaction partner moiety combinations are possible, and these examples are in no way limiting. The moieties of the modulating modifier of Formula 5 configured to support cell growth, viability, transplantability, or any combination thereof, may comprise: alkyl or fluoroalkyl (which includes perfluoroalkyl) moieties; mono- Saccharides or polysaccharides (which may include, but are not limited to, polydextrose); alcohols (which include, but are not limited to, propargyl alcohol); polyols, which include, but are not limited to, polyvinyl alcohol; alkylene ethers, It includes, but is not limited to, polyethylene glycol; polyelectrolytes (which include, but are not limited to, polyacrylic acid or polyvinylphosphonic acid); amine groups (which include, but are not limited to, derivatives thereof, such as, but are not limited to, alkylated amine groups , hydroxyalkylated amine groups, guanidine salts, and heterocyclic groups containing an unaromatic nitrogen ring atom, such as (but not limited to) morpholinyl or piperazinyl); carboxylic acids, including but not limited to propylene alkynoic acids (which can provide a carboxylate anion surface); phosphonic acids, including but not limited to ethynylphosphonic acids (which can provide a phosphonate anion surface); sulfonic acid anions; carboxybetaines; sulfobetaines; amines Sulfonic acid; or amino acid. The moiety that provides the modulating modifier of formula 5 that enhances cell growth, viability, transplantability, or any combination thereof, can be linked directly (L', where m=0) or indirectly to the reaction partner moiety R _px. When the reaction is paired with the moiety R _pxWhen indirectly linked to a moiety that provides enhanced cell growth, viability, transplantability, or any combination thereof, the reactive partner moiety R _pxCan be connected to a one-key connector L' (m=1). reaction pairing part R _pxA moiety that can be attached to a first end of the linker L', and that provides enhanced cell growth, viability, transplantability, or any combination thereof, can be attached to a second end of the linker L'. The linker L' may have a linear moiety, wherein a main chain of the linear moiety may comprise a main chain selected from the group consisting of silicon atoms, carbon atoms, nitrogen atoms, oxygen atoms, sulfur atoms and phosphorus atoms (subject to chemistry known in the art). 1 to 100 non-hydrogen atoms in any combination of bonding limits). In some non-limiting examples, the linkage L' can be interrupted using any combination of ether, amine, carbonyl, amido, or phosphonate groups. In addition, the linker L' may have one or more arylidene, heteroaryl or heterocyclic groups that interrupt the main chain of the linker L'. In some embodiments, the backbone of the linker L' may contain 10 to 20 atoms. In other embodiments, the backbone of the linker L' may comprise about 5 atoms to about 100 atoms, about 10 atoms to about 80 atoms, about 10 atoms to about 50 atoms, or about 10 atoms to about 40 atoms. In some embodiments, the backbone atoms are all carbon atoms. In other embodiments, the backbone atoms are not all carbon, but may include any feasible combination of silicon, carbon, nitrogen, oxygen, sulfur, or phosphorus atoms subject to chemical bonding constraints known in the art . When the conditioning modifier (Formula 5) reacts with a surface having a surface modifying ligand (Formula 3), a substrate having a conditioning surface of Formula 2 is formed. Then, the linker L' and the linker L'' are formally part of the linker L, and the reaction pairing part R _pxwith reactive moiety R _xThe reaction yields the coupling group CG of formula 2. surface modifierThe surface modifier system has LG-(L'') _j-R _xA compound of one of the structures of (Formula 4). The linking group LG is covalently linked to the oxide on the surface of the DEP substrate. The dielectrophoretic substrate can be silicon or aluminum oxide, and the oxide can be present as part of the substrate's natural chemical structure or can be introduced as discussed herein. The linking group LG may be a siloxane or phosphonate group formed by the reaction of a siloxane or phosphonate group with an oxide on the surface of the substrate. Reaction part R _xas described above. Reaction part R _xThe linkage to the linking group LG can be directly (L'', j=0) or indirectly via a linkage L'' (j=1). The linking group LG can be attached to one of the first ends of the linker L'' and the reactive moiety R _xCan be attached to a second end of the linker L", which will be at the distal end of the surface of the substrate after the surface-modifying ligand has been attached to the surface as in Formula 3.

Equation 4 Equation 3 The linker L'' may have a straight-chain moiety, wherein one of the main chains of the straight-chain moiety comprises 1 to 100 atoms selected from any combination of silicon atoms, carbon atoms, nitrogen atoms, oxygen atoms, sulfur atoms, and phosphorus atoms non-hydrogen atoms. In some non-limiting examples, any combination of ether, amine, carbonyl, amido, or phosphonate groups can be used to interrupt the linker L''. In addition, the linker L'' may have one or more arylidene groups, heteroarylidene groups or heterocyclic groups that interrupt the main chain of the linker L''. In some embodiments, the backbone of the linker L″ may contain 10 to 20 atoms. In other embodiments, the backbone of the linker L″ may comprise about 5 atoms to about 100 atoms, about 10 atoms to about 80 atoms, about 10 atoms to about 50 atoms, or about 10 atoms atom to about 40 atoms. In some embodiments, the backbone atoms are all carbon atoms. In other embodiments, the backbone atoms are not all carbon, but may include any feasible combination of silicon, carbon, nitrogen, oxygen, sulfur, or phosphorus atoms subject to chemical bonding constraints known in the art . cleavable partIn various embodiments, any of the following may further comprise a cleavable moiety (as will be discussed below): a moiety that supports cell growth, viability, portability or any combination thereof, a linker L, a linker Sub L', linking sub L'' or coupling group CG. The lysable moiety can be configured to allow destruction of a conditioning surface of a microfluidic device, which promotes the implantability of one or more biological cells. In some embodiments, the portability of one or more biological cells may be desirable to be able to move the cells after culturing the cells for a period of time and especially to export the cells from a microfluidic device. Composition of substratesAccordingly, a composition is provided comprising: a substrate having a dielectrophoretic (DEP) configuration and a surface; and a conditioning surface covalently bonded to oxide moieties of the surface of the substrate. The conditioning surface on the substrate can have one of the structures of Equation 1 or Equation 2:

; Equation 1 Equation 2 Among them: LG is a linking group; L is a linker, which may be present (n=1) or absent (n=0); part of it supports cell growth, viability, transplantability in microfluidic devices part of sex or any combination thereof; and CG is a coupling group, as defined herein. The conditioning surface may comprise a linking group LG covalently bonded to the oxide moiety of the surface. The linking group can be further linked to a moiety that is configured to support cell growth, viability, portability, or any combination thereof. The linking group may be a siloxy linking group. In other embodiments, the linking group can be a phosphonate group. The linking group can be linked directly or indirectly to the moiety that is configured to support cell growth, viability, transplantability, or any combination thereof. The linking group can be indirectly linked to the moiety configured to support cell growth, viability, transplantability, or any combination thereof, via attachment to a first end of a linker. The linker may further comprise a linear moiety, wherein a main chain of the linear moiety may have 1 to 200 atoms selected from any combination of silicon atoms, carbon atoms, nitrogen atoms, oxygen atoms, sulfur atoms and phosphorus atoms Non-hydrogen atoms, as discussed above. The backbone of the linear moiety may further comprise one or more arylidene moieties. The linker may have a coupling group CG as defined above. The coupling group CG may contain a triazolyl moiety. The triazolyl moiety may interrupt the linear portion of the linker or may be attached to a second end of the linear portion of the linker. The second end of the linker may be located at the distal end of the surface of the substrate. Moieties that are configured to support cell growth, viability, transplantability, or any combination thereof, may include: alkyl or fluoroalkyl (which includes perfluoroalkyl) moieties; monosaccharide or polysaccharide (which may include ( but not limited to) polydextrose); alcohols (including but not limited to propargyl alcohol); polyols, including but not limited to polyvinyl alcohol; alkylene ethers, including but not limited to polyvinyl alcohol ethylene glycol; polyelectrolytes (which include, but are not limited to, polyacrylic acid or polyvinylphosphonic acid); amines (which include derivatives thereof, such as, but not limited to, alkylated amines, hydroxyalkylated amines, Guanidine salts and heterocyclic groups containing an unaromatic nitrogen ring atom such as, but not limited to, morpholinyl or piperazinyl); carboxylic acids, including but not limited to propyonic acid (which can provide carboxylic acids anionic surfaces); phosphonic acids including, but not limited to, ethynylphosphonic acids (which can provide phosphonic acid anionic surfaces); sulfonic acid anions; carboxybetaines; sulfobetaines; amine sulfonic acids; or amino acids. In some embodiments, a mixture of different moieties, such as, but not limited to, a mixture of anionic functional groups and a mixture of cationic functional groups that provide a zwitterionic modulating surface, is incorporated into the modulating surface. The conditioning surface may contain alkyl or perfluoroalkyl moieties. The alkyl or perfluoroalkyl moieties can have a backbone length of greater than 10 carbons. The conditioning surface may comprise carbohydrate moieties, and may be polydextrose. In other embodiments, the conditioning surface may comprise alkylene ether moieties. The isoalkylene ether moiety may be polyethylene glycol. The conditioning surface may further comprise a cleavable moiety. The cleavable moiety can be configured to allow disruption of the regulatory surface, thereby promoting the transplantability of one or more biological cells. Another composition is provided comprising: a substrate comprising a dielectrophoresis (DEP) configuration and a surface; and a surface modifying ligand covalently bonded to an oxide moiety of the surface of the substrate. The substrate with a surface-modified ligand can have a structure of formula 3:

Formula 3 Among them: LG is a linking group; L'' is an optional linker, and j is 0 or 1. The linker L'' exists when j=1, and the linker L'' does not exist when j=0; and R _xis a reaction moiety as described herein. The reactive moiety of the surface-modified ligand can be an azide, amine, bromo, thiol, active ester, succinimidyl, or alkynyl moiety. The surface-modifying ligands can be covalently linked to the oxide moiety via a linking group. The linking group may be a siloxy moiety. In other embodiments, the linking group can be a phosphonate group. The linking group can be indirectly attached to the reactive moiety of the surface-modifying ligand via a linker. The linking group can be attached to a first end of the linker and the reactive moiety can be attached to a second end of the linker. The linker L'' may comprise a straight-chain moiety, wherein one of the main chains of the straight-chain moiety comprises 1 to 100 atoms selected from any combination of silicon atoms, carbon atoms, nitrogen atoms, oxygen atoms, sulfur atoms, and phosphorus atoms non-hydrogen atoms. The main chain of the linker L'' may contain 10 to 20 atoms. In other embodiments, the backbone of the linker L'' may contain from about 5 atoms to about 50 atoms. In some embodiments, the backbone of the linker L'' may be all carbon atoms. The backbone of the linear moiety may contain one or more arylidene moieties. The linker L'' may contain a triazolyl moiety. The triazolyl moiety can interrupt the linker L″ or can be attached at one of the ends of the linker L″. The surface-modifying ligand may contain a cleavable moiety. The lysable moiety can be configured to allow destruction of a conditioning surface of a microfluidic device, thereby promoting the implantability of one or more biological cells. method of preparationIn some embodiments, chemical vapor deposition is used to deposit conditioning surfaces or surface-modifying ligands on interior surfaces of the microfluidic device. Modified surface/surface-modified ligands can be achieved by vapor deposition of molecules to achieve close-packed monolayers, including covalent bonding of molecules of modified surface/surface-modified ligands to microfluidic devices (100, 200, 240 , 290, 400, 500A to 500E, 600) on the inner surface of the molecule. To achieve a desired bulk density, vapor deposition including, for example, an Molecules of the group-terminated siloxane for a period of at least 15 hours (eg, at least 20, 25, 30, 35, 40, 45, or more than 45 hours). This vapor deposition is typically performed under vacuum and in the presence of a source of water, such as an aqueous sulfate (eg, MgSO ₄·7H ₂0). Generally, the increased temperature and duration of vapor deposition results in improved properties for tuning the surface/surface-modifying ligands. In some embodiments, surface modulating or surface modifying ligands can be introduced by reaction in a liquid phase. To prepare the microfluidic surface, the lid, microfluidic circuit material, and electrode active substrate can be treated by an oxygen plasma treatment that removes various impurities while introducing an oxidized surface (eg, as described herein). generally covalently modified surface oxides). The oxygen plasma cleaner can be operated, for example, at 100 W under vacuum for 60 seconds. Alternatively, a liquid phase treatment can be used, which includes an oxidizing agent, such as hydrogen peroxide, to oxidize the surface. For example, a mixture of hydrochloric acid and hydrogen peroxide or a mixture of sulfuric acid and hydrogen peroxide (eg, a piranha solution, which may have a ratio of sulfuric acid to hydrogen peroxide ranging from about 3:1 to about 7:1) . For example, the vapor deposition process can be optionally modified by pre-cleaning the lid, microfluidic circuit material, and electrode active substrate. For example, the pre-cleaning may comprise a solvent bath, such as an acetone bath, an ethanol bath, or a combination thereof. The solvent bath may contain sonication. In some embodiments, vapor deposition is used to coat the inner surface(s) of the microfluidic device after the microfluidic device has been assembled to form an enclosure defining a microfluidic circuit. When a substrate with a surface-modifying ligand is further reacted with a conditioning modifier to produce a substrate with a conditioning surface, the reaction can be performed in situ using any suitable solvent that will dissolve the reagent and not Destruction of microfluidic circuit materials or surfaces with a surface modified ligand. In some embodiments, the solvent is an aqueous solution. Method for preparing a conditioning surface or a surface comprising a surface-modifying ligandAccordingly, a method for preparing a modified surface of a microfluidic device having a dielectrophoresis (DEP) configuration is provided, comprising the steps of: providing a surface of a substrate of a microfluidic device, wherein the substrate comprises a DEP topography; reacting the oxide of the surface with a modifier, thereby converting the surface of the substrate into a modified surface. In some embodiments, the surface of the substrate may be plasma cleaned to provide oxide on the surface. In some embodiments, the surface can be plasma cleaned prior to assembly of the microfluidic device. In other embodiments, the surface may be plasma cleaned after assembling the microfluidic device. The step of the method of reacting the oxide of the surface with the modifier is performed by exposing the surface to a liquid that includes the modifier. In some embodiments, the step of reacting the oxides of the surface may be performed by exposing the surface to a vapor containing a modifier at a reduced pressure. In some embodiments, the modifier may comprise a surface modulating agent having a first moiety configured to covalently react with the surface and configured to support cell growth, viability, transplantability, or the like A second part of any combination, whereby the surface is modified to a surface that is conditioned to support cell growth, viability, transplantability, or any combination thereof. The surface-modifying agent may have one of the structures of Formula 6:

Formula 6 The first part may contain a linking group LG, which may be a siloxane or phosphonic acid group. The linking group LG can be linked directly or indirectly to a moiety configured to support cell growth, viability, transplantability, or any combination thereof. The first moiety can be directly (n=0) or indirectly (n=1) linked to a second moiety configured to support cell growth, via attachment to a first end of a linkage L, Part of viability, portability, or any combination thereof. The linker L may further comprise a linear moiety, wherein a main chain of the linear moiety may have 1 to 200 atoms selected from any combination of silicon atoms, carbon atoms, nitrogen atoms, oxygen atoms, sulfur atoms and phosphorus atoms non-hydrogen atoms. The backbone of the linear moiety may further comprise one or more arylidene moieties. The second moiety ("moiety") of the surface modulating agent may comprise: an alkyl or fluoroalkyl (which includes perfluoroalkyl) moieties; a monosaccharide or polysaccharide (which may include, but is not limited to, polydextrose); alcohols (which includes, but is not limited to, propargyl alcohol); polyols, which include, but are not limited to, polyvinyl alcohol; alkylene ethers, which include, but are not limited to, polyethylene glycols; polyelectrolytes (which include ( but not limited to) polyacrylic acid or polyvinylphosphonic acid); amine groups (including derivatives thereof, such as (but not limited to) alkylated amine groups, hydroxyalkylated amine groups, guanidine salts and containing an unaromatic nitrogen ring Heterocyclyl groups of atoms, such as, but not limited to, morpholinyl or piperazinyl); carboxylic acids, including but not limited to propyolic acid (which can provide a carboxylate anion surface); phosphonic acids, including ( but not limited to) ethynylphosphonic acid (which can provide a phosphonic acid anion surface); sulfonic acid anion; carboxybetaine; sulfobetaine; sulfamic acid; or amino acid. The second moiety of the surface modifying agent may comprise an alkyl or perfluoroalkyl moiety. The alkyl or perfluoroalkyl moieties can have a backbone length of greater than 10 carbons. The second portion of the surface modulating agent may comprise a carbohydrate moiety, and may be polydextrose. In other embodiments, the second moiety of the surface modifying agent may comprise an alkylene ether moiety. The isoalkylene ether moiety may be polyethylene glycol. The second portion of the surface modulating agent may further comprise a cleavable moiety, which may be located within the linker L or may be part of the second portion of the surface modulating agent. The cleavable moiety can be configured to allow disruption of the regulatory surface, thereby promoting the transplantability of one or more biological cells. In various embodiments, the modifier may comprise a surface modifier having a structure of Formula 4 as defined above, wherein the surface modifier comprises: a first moiety LG configured to interact with the surface reaction; and a second part R _x, which can be a reactive moiety or can be modified to include a reactive moiety comprising an azide, amine, bromo, thiol, active ester, succinimidyl, or alkynyl moiety) , thereby converting the surface to a surface comprising a surface-modifying ligand having a structure of formula 3, as described above. In some embodiments, the first portion of the surface modifier, which is configured to react with the oxide of the surface, can be a siloxane or a phosphonic acid. In some embodiments, the method includes the step of reacting a surface comprising a surface-modifying ligand (Formula 3) with a modulating modifier comprising a modulating modifier that is configured to support cell growth, storage A first moiety of activity, portability, or any combination thereof, and a second moiety R configured to react with the reactive moiety of the surface-modifying ligand _px; thereby providing a surface configured to support cell growth, viability, transplantability, or any combination thereof of a biological cell, the surface having a structure of Formula 2, as described above. The conditioning modifier may have one of the structures of Formula 5. In some embodiments, the first moiety of the conditioning modifier includes at least one of the following: an alkylene oxide moiety, an amino acid moiety, a carbohydrate moiety, an anionic moiety, a cationic moiety, and a zwitterionic moiety. In various embodiments, any of the surface conditioning agent, surface modifier, or conditioning modifier can further comprise a cleavable moiety as described herein. Conditioning surfaces with other componentsThe conditioning surface may comprise other components than a polymer formed by a covalently linked moiety or a conditioning surface, or the conditioning surface may comprise a polymer or a conditioning surface formed by a covalently linked moiety, Additional components may also be included, the covalently linked moieties comprising biocompatible metal ions (eg calcium, sodium, potassium or magnesium), antioxidants, surfactants and/or essential nutrients. A non-limiting exemplary list includes: vitamins such as B7, alpha-tocopherol, alpha-tocopheryl acetate, vitamin A and its acetate; proteins such as BSA, catalase, insulin, transferrin , superoxide dismutase; small molecules such as corticosterone, D-galactose, ethanolamine hydrochloride, reduced glutathione, L-carnitine hydrochloride, linoleic acid, linolenic acid, progesterone, putrescine dihydrochloride and triiodothyronine; and salts including, but not limited to, sodium selenite, sodium phosphate, potassium phosphate, calcium phosphate and/or magnesium phosphate. Antioxidants may include, but are not limited to, carotenoids, cinnamic acid and its derivatives, ferulic acid, polyphenols such as flavonoids, quinones and their derivatives (which include mitoxantrone-Q), N- Acetylcysteine and antioxidant vitamins (such as ascorbic acid, vitamin E and the like). The conditioning surface may comprise a medium supplement (such as B-27® supplement) containing antioxidants and many of the other components listed above. B-27® Supplement is a commercially available (50x) serum-free medium from ThermoFisher Scientific (Cat# 17504044). In some embodiments, the at least one modulating surface may comprise one or more components of mammalian serum. In some embodiments, the mammalian serum is fetal bovine serum (FBS) or calf serum (FCS). The modulating surface may comprise specific components of mammalian serum (such as specific amounts and types of proteins commonly found in serum), which may be provided in defined amounts or types from serum-free media or synthetic media. In other embodiments, the at least one modulating surface does not comprise a mammalian serum. In various embodiments, at least one conditioning surface may not contain any titanium, nickel or iron metal ions. In other embodiments, at least one conditioning surface may not contain any significant concentration of titanium, nickel or iron metal ions. In other embodiments, the at least one conditioning surface may not contain any gold, aluminum or tungsten metal ions. Treatment with reagents to reduce adhesion / mixture of reagentsWhen cells are cultured within a microfluidic device, the cells actively secrete proteins and other biomolecules and passively shed similar biomolecules that can adhere to surfaces within the microfluidic device. Cultured cells can adhere to each other or to the conditioning surface and become difficult to remove from the growth chamber for export from the microfluidic device. Additionally, in some cases, it may be desirable to bring additional cell types into the microfluidic device, the same as or different from the cultured cells. These newly delivered cells can also become adherent to surface fouling accumulated within the microfluidic environment and are difficult to remove from the device at a subsequent point in time. Treatment with proteases such as trypsin or Accutase® (an enzymatic mixture with proteolytic and collagenolytic activity, Innovative Cell Technologies) does not provide sufficient efficacy (for a non-limiting example) to allow adherent cells from microfluidics device output performance. A mixture of one or more proteins and/or peptides that provide anti-adhesion properties can act to reduce adhesion in both cases. Biomolecules or small molecules that have activity against one of the various cell adhesion mechanisms can be used. Some cell adhesion mechanisms that can be inhibited can be active actin fibril formation and related programs, which can be inhibited by small molecules using compounds such as cytochalasin B (New England Biosciences Cat No: M0303S), a microfibril extension agent) to achieve. Specific receptor-driven adhesion programs, such as, but not limited to, inhibition of integrin receptor-mediated adhesion to fibronectin (which can be found on a stained surface) can be achieved by using RGD-containing peptides. Another type of fouling material (ie, nucleic acids released from dead cells) can attract cell binding, which can be achieved by using an endonuclease that will cleave the fouling nucleic acid. A specific endonuclease (DNase 1 (DNase 1, Sigma Aldrich, Catalog No. AMPD1-1KT)) also binds to actin to thus provide a dual-active block of adhesion. In some embodiments, a mixture of one of all three blocking agents can be used to prevent/reduce cell adhesion. General Processing Agreement / After cultivation:For cells that have grown within a microfluidic device for 2, 3, 4, or more days, a mixture of three anti-adherence agents or a single anti-adherence agent (as will be described below) can be flowed into the microfluidic device and are allowed to diffuse into the growth chamber for a time period of about 20 minutes, about 30 minutes, about 40 minutes, about 50 minutes, or about 60 minutes before exporting the cells. preprocessingFor cells to be imported into a microfluidic device, the cells can be pre-incubated in a medium containing the mixture or a single anti-adherent for about 30 minutes, and then the cells are imported into the microfluidic chip. Inhibition continued for a time period of 1 hour, 2 hours, 3 hours, or more than 3 hours without further addition of reagents. The RGD tripeptide (mw. 614.6, Santa Cruz Biotechnology Cat No: sc-201176) can be present in the medium or pre-infused medium at a concentration of one of about 0.1 mM to about 20 mM. In some embodiments, the RGD tripeptide can be about 0.1 mmol, about 0.5 mmol, about 0.7 mmol, about 1.0 mmol, about 3.0 mmol, about 5.0 mmol, about 8.0 mmol It is present at a concentration of auricular, about 10.0 mmol, or any value within the range. Cytochalasin B can be about 0.01 mmol or about 50 mmol or about 0.01 mmol, about 0.05 mmol, about 0.1 mmol, about 2 mmol, about 4 mmol, about 6 mmol A concentration of one of molar, about 8 mmol, about 10 mmol, about 20 mmol, about 30 mmol, about 50 mmol, or any value within the range is present in the pre-infusion medium. DNase 1 can be from about 0.001 U/microliter to about 10 U/microliter or about 0.001 U/microliter, about 0.005 U/microliter, about 0.01 U/microliter, about 0.05 U/microliter, about 1.0 U/microliter It is present at a concentration of one of liters, about 5.0 U/microliter, about 10 U/microliter, or any value within the range. In some embodiments, a single agent can be used to reduce adhesion before cells have been cultured in a microfluidic device or after cells have been cultured in a microfluidic device. For example, the RGD tripeptide can be used for pre-incubation at a concentration of 5 mg/ml or can be flowed into the microfluidic device as a treatment agent prior to export. Another inhibitor that can be used is the tetrapeptide fibronectin inhibitor (Arg-Gly-Asp-Ser-OH, mw. 433.4, Santa Cruz Biotechnology Cat No: sc-202156). The fibronectin inhibitor can be used at a concentration of about 1.75 micrograms/ml (4 micromoles). Similar to the use of protein or small molecule agents to reduce or prevent adhesion, antibodies to extracellular adhesion-related proteins can be used to affect output and portability within microfluidic devices. A non-limiting example is anti-B1 integrin: Colony M-106 (Santa Cruz Biotechnology Cat No: sc-8978). Conditioning surface containing a cleavable moietyIn some embodiments, the modulating surface can have a cleavable moiety incorporated within a covalently or non-covalently linked molecule of the modulating surface. The regulatory surface may comprise a peptide motif (such as RGD) that has one of the functions described above, or it may have another peptide motif that promotes cell growth or provides contact cues for cell proliferation. In other embodiments, the conditioning surface provides non-specific support to cells and may only serve to buffer cells from the silicon or aluminum oxide surface of the microfluidic device. It may be desirable to destroy the conditioning surface after completing a cycle of cell culture to facilitate the output of the expanded cell population within a growth chamber of the microfluidic device. This can be used when cells exhibit adhesive behavior. The regulatory surface can be destroyed, partially or fully removed, by incorporating other peptide motifs, which are the enzymatic bases of a protease that is not highly secreted by the cell of interest. In a non-limiting example, the peptide motif of ENLYQS (Glu-Asn-Leu-Tyr-Gln-Ser) can be incorporated into a modulating surface at pre-designed time intervals. This motif is one of the enzyme bases of the TEV protease (tobacco etch virus cysteine protease, Sigma Aldrich catalog no. T4455) that is highly sequence specific and therefore used for highly controlled cleavage. After completing the incubation cycle, the TEV protease can flow into the microfluidic device and be allowed to diffuse into isolated regions of the growth chamber. Next, the conditioning surface is disrupted to facilitate export of cells within the microfluidic device. Accordingly, various other proteolytic motifs can be designed and incorporated into a regulatory surface to be cleaved by one suitable for a particular protease that can be devised by those skilled in the art. fluid mediumWith regard to the above discussion in relation to microfluidic devices having a channel and one or more growth chambers, a fluid medium (eg, a first medium and/or a second medium) may be capable of maintaining a cell in a substantially viable any fluid in a changing state. This variable state will depend on the biological micro-object and the culture experiment performed. The first fluid medium and/or the second fluid medium can provide both the fluid and dissolved gas components required for cell survival, and a buffered fluid medium or pH monitoring or both can also be used to maintain pH within a desired range Inside. If the cell line is a mammalian cell, the first fluid medium and/or the second fluid medium may comprise mammalian serum known in the art or a serum-free synthetic medium capable of providing essential nutrients, endocrine, growth factors or cell growth signals. Similar to the conditioning surfaces described above, the first fluid medium and/or the second fluid medium may comprise fetal bovine serum (FBS) or calf serum (FCS). Alternatively, the first fluid medium and/or the second fluid medium may not contain any animal-derived serum, but may contain a synthetic medium, which may contain physiologically relevant metal ions including but not limited to sodium, potassium, calcium, magnesium and/or zinc) any or all of antioxidants, surfactants and/or essential nutrients. Synthetic media can be serum-free, while still containing some proteins, in defined amounts and types. A non-limiting exemplary list of components in a serum-free medium includes: vitamins such as B7, alpha-tocopherol, alpha-tocopherol acetate, vitamin A and its acetates; proteins such as BSA, Catalase, insulin, transferrin, superoxide dismutase; small molecules such as corticosterone, D-galactose, ethanolamine hydrochloride, reduced glutathione, L-carnitine hydrochloride, linseed oil acid, linolenic acid, progesterone, putrescine dihydrochloride and triiodothyronine; and salts including, but not limited to, sodium selenite, sodium phosphate, potassium phosphate, calcium phosphate and/or Magnesium Phosphate. The fluid medium may contain any of the antioxidants described above for conditioning surfaces. Fluid media can be sterile filtered through a 0.22 micron filter unit (VWR, Cat. No. 73520-986). In some embodiments, a suitable medium can comprise or consist entirely of any of the following: Dulbecco's Modified Eagle's Medium (ThermoFisher Scientific, Cat # 11960-051), FreeStyle ^TMMedia (Invitrogen, ThermoFisher Scientific, Cat. No. 11960-051), RPMI-1640 (GIBCO®, ThermoFisher Scientific, Cat. No. 11875-127), Hybridoma-SFM (ThermoFisher Scientific, Cat. No. 12045-076) , Medium E (Stem Cell, Cat. No. 3805), 1X CD CHO medium (ThermoFisher Scientific, Cat. No. 10743-011), Iscove modified Dulbecco’s medium (ThermoFisher Scientific, Cat. No. 12440- 061) or CD DG44 medium (ThermoFisher Scientific, Cat. No. 10743-011). The medium may additionally comprise fetal bovine serum (FBS, available from GIBCO®, ThermoFisher Scientific), heat-inactivated fetal bovine serum or calf serum (FCS, Sigma-Aldrich Cat Nos. F2442, F6176, F4135 and others). FBS can be present at one of the following concentrations: about 1% v/v to about 20% v/v, about 1% v/v to about 15% v/v, about 1% v/v to about 10% v/v, or From about 1% v/v to about 5% v/v, or any value within any of those ranges. The medium may additionally contain human AB serum (Sigma-Aldrich, Cat. No. S2146) and may be present at one of the following concentrations: about 1% v/v to about 20% v/v, about 1% v/v to about 15% v/v, from about 1% v/v to about 10% v/v, or from about 1% v/v to about 5% v/v, or any value within any of these ranges. The medium may additionally contain penicillin-streptomycin (ThermoFisher Scientific, Cat. No. 15140-163). Streptococcus can be present at a concentration in one of the following ranges: about 0.01% v/v to about 10% v/v, about 0.1% v/v to about 10% v/v, about 0.01% v/v to about 5% v/v, about 0.1% v/v to about 5% v/v, about 0.1% v/v to about 3% v/v, about 0.1% v/v to about 2% v/v, about 0.1% v /v to about 1% v/v or any value within any of these ranges. In other embodiments, the culture medium may comprise Geneticin (ThermoFisher Scientific, Cat. No. 101310-035). Geneticin may be present at one of the following concentrations: about 0.5 micrograms/ml, about 1.0 micrograms/ml, about 5.0 micrograms/ml, about 10.0 micrograms/ml, about 15 micrograms/ml, about 20 micrograms/ml, about 30 micrograms/ml ml, about 50 micrograms/ml, about 70 micrograms/ml, about 100 micrograms/ml, or any value within these ranges. The medium may contain a buffer. The buffer can be one of the Good buffers. The buffer may be, but is not limited to, (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid) (HEPES) (ThermoFisher Scientific, Cat. No. 15630-080). The buffer may be present at one of the following concentrations: about 1 mM, about 3 mM, about 5 mM, about 7 mM, about 9 mM, about 10 mM, about 12 mM ear, about 15 mmol, about 20 mmol, about 40 mmol, about 60 mmol, about 100 mmol, or any value within these ranges. The medium may additionally contain the dipeptide substitute for glutamine, GlutaMAX ^TM(GIBCO® ThermoFisher Scientific, Cat No. 35050-079). The substitute for glutamine can be present in one of the following concentrations: about 0.2 mmol, about 0.5 mmol, about 0.7 mmol, about 1.0 mmol, about 1.2 mmol, about 1.5 mmol, About 1.7 mmol, about 2.0 mmol, about 2.5 mmol, about 3.0 mmol, about 4.0 mmol, about 7.0 mmol, or about 10.0 mmol, or any value within these ranges. The medium may contain MEM non-essential amino acids (ThermoFisher Scientific, Cat. No. 10370-088). The MEM non-essential amino acid can be present at one of the following concentrations: about 0.2 mmol, about 0.5 mmol, about 0.7 mmol, about 1.0 mmol, about 1.2 mmol, about 1.5 mmol, about 1.7 mmol, about 2.0 mmol, about 2.5 mmol, about 3.0 mmol, about 4.0 mmol, about 7.0 mmol, or about 10.0 mmol, or any value within these ranges. The medium may additionally contain glucose (ThermoFisher Scientific, Cat. No. 15023-021). Glucose can be present at one of the following concentrations: about 0.1 g/L, about 0.3 g/L, about 0.5 g/L, about 0.8 g/L, about 1.0 g/L, about 1.5 g/L, about 2.0 g/L, About 2.5 g/L, about 3.0 g/L, about 3.5 g/L, about 4.0 g/L, about 5.0 g/L, about 7.0 g/L, about 10.0 g/L, or any value within these ranges. The medium may additionally contain mercaptoethanol (ThermoFisher Scientific, Cat. No. 31350-010). The mercaptoethanol can be present at one of the following concentrations: about 0.001% v/v to about 1.5% v/v, about 0.005% v/v to about 1.0% v/v, about 0.01% v/v to about 1.0% v/v , about 0.15% v/v to about 1.0% v/v, about 0.2% v/v to about 1% v/v, or any value within these ranges. The medium may contain OPI medium supplements containing oxaloacetic acid, pyruvate and insulin (Sigma-Aldrich, Cat. No. O-5003). The OPI medium supplement can be present at one of the following concentrations: about 0.001% v/v to about 1.5% v/v, about 0.005% v/v to about 1.0% v/v, about 0.01% v/v to about 1.0% v /v, about 0.15% v/v to about 1.0% v/v, about 0.2% v/v to about 1% v/v, or any value within these ranges. The medium may contain B-27 supplement (50x), serum-free medium (ThermoFisher Scientific, Cat. No. 17504-163). The B-27 supplement can be present at one of the following concentrations: about 0.01% v/v to about 10.5% v/v, about 0.05% v/v to about 5.0% v/v, about 0.1% v/v to about 5.0% v/v, about 0.5% v/v to about 5% v/v, or any value within these ranges. As described herein, a medium or a supplement to a medium can include one or more Pluronic® polymers for producing a conditioning surface, and can include Pluronic® L44, L64, P85, F68, and F127 (which include F127NF). The Pluronic® polymer can be present in the medium at one of the following concentrations: about 0.001% v/v to about 10% v/v, about 0.01% v/v to about 5% v/v, about 0.01% v/v to about 1% v/v or about 0.05% v/v to about 1% v/v. For medium supplements that may be provided as one of a set, the concentration may be 1, 5, 10, 100 or about 100 times the final medium concentration. The medium may contain IL6 (Sigma-Aldrich, Cat. No. SRP3096-20UG). IL 6 can be present at one of the following concentrations: about 1 nM, about 5 nM, about 10 nM, about 15 nM, about 20 nM, about 25 nM, about 30 nM, about 40 nM, about 50 nM, or within these ranges any value. The medium may additionally contain sodium pyruvate (ThermoFisher Scientific, Cat. No. 11360-070). The substitute for glutamine can be present in one of the following concentrations: about 0.1 mmol, about 0.02 mmol, about 0.04 mmol, about 0.06 mmol, about 0.08 mmol, about 0.1 mmol, about 0.5 mmol, about 0.7 mmol, about 1.0 mmol, about 1.2 mmol, about 1.5 mmol, about 1.7 mmol, about 2.0 mmol, about 2.5 mmol, about 3.0 mmol, about 4.0 mmol, about 7.0 mmol, or about 10.0 mmol, or any value within these ranges. Gas environmentThe system provides a mixture of gases required for cell survival, including, but not limited to, oxygen and carbon dioxide. Both gases dissolve into the fluid medium and are available to the cells, thus changing the gas content of the fluid medium in an isolated region of a growth chamber over time. In particular, carbon dioxide content can vary over time, which affects the pH of the fluid medium in the microfluidic device. In some experimental conditions, non-optimal oxygen partial pressures may be used. temperature controlIn some embodiments, the at least one conditioning surface of the growth chamber(s) and/or the flow region(s) is conditioned by controlling the temperature of the at least one conditioning surface. The system can include a component that can control and modulate the temperature of at least one conditioned surface of the growth chamber and/or flow region of the microfluidic device. The system may include Peltier heating, resistive heating, or any other suitable method for providing temperature modulation to the microfluidic device. The system may also include sensors and/or feedback components to control the heat input to a predetermined range. In some embodiments, the at least one conditioning surface has at least about 25°C, about 26°C, about 27°C, about 28°C, about 29°C, about 30°C, about 31°C, about 32°C One of C, about 33°C, about 34°C, about 35°C, about 36°C, about 37°C, about 38°C, about 39°C, or about 40°C and stable at that temperature place. In some embodiments, at least one surface has a temperature greater than about 25°C. In other embodiments, at least one surface has a temperature ranging from about 30°C to about 40°C, from about 35°C to about 38°C, or from about 36°C to about 37°C. In some embodiments, the at least one conditioning surface has a temperature of at least about 30°C. Flow controller to provide perfusion during incubationThe flow controller can perfuse the first fluid medium in the flow region (as described above) during the incubation cycle to provide nutrients to cells in the growth chamber and remove waste from the growth chamber, where the exchange of nutrients and waste The removal occurs substantially via diffusion. The controller may be a separate component from the microfluidic device or may be incorporated as part of the microfluidic device. The flow controller can be configured to discontinuously perfuse the medium into the flow area. The flow controller can be configured to perfuse the medium(s) in a periodic pattern or in an irregular pattern. In some other embodiments, the controller may be configured to operate approximately every 4 hours, 3 hours, 2 hours, 60 minutes, 57 minutes, 55 minutes, 53 minutes, 50 minutes, 47 minutes, 45 minutes, 43 minutes, 40 minutes, 37 minutes, 35 minutes, 33 minutes, 30 minutes, 27 minutes, 25 minutes, 23 minutes, 20 minutes, 17 minutes, 15 minutes, 13 minutes, 10 minutes, 7 minutes, or 5 minutes Pour into the flow area once. In some embodiments, the controller can be configured to perfuse the fluid medium about every 5 minutes to about every 20 minutes. In other embodiments, the controller can be configured to perfuse the fluid medium about every 15 minutes to about every 45 minutes. In other embodiments, the controller can be configured to perfuse the fluid medium every 30 minutes to about every 60 minutes. In other embodiments, the controller can be configured to perfuse the fluid medium every 45 minutes to about every 90 minutes. In some other embodiments, the controller can be configured to perfuse the fluid medium every 60 minutes to about 120 minutes. Alternatively, the controller can be configured to perfuse the fluid medium every 2 hours to every 6 hours. In some embodiments, the controller 226 may be configured to operate at a time of about 5 seconds, about 10 seconds, about 15 seconds, about 20 seconds, about 25 seconds, about 30 seconds, about 35 seconds, about 40 seconds, about The medium is perfused for a time period of one of 45 seconds, about 50 seconds, about 55 seconds, about 60 seconds, about 65 seconds, or about 70 seconds. In other embodiments, the controller can be configured to perfuse the culture medium for about 1 minute, about 1.2 minutes, about 1.4 minutes, about 1.5 minutes, about 1.6 minutes, about 1.8 minutes, about 2.0 minutes, about 2.2 minutes, about 2.4 minutes minutes, about 2.5 minutes, about 2.6 minutes, about 2.8 minutes, about 3.0 minutes, about 3.2 minutes, about 3.4 minutes, about 3.5 minutes, about 3.6 minutes, about 3.8 minutes, or about 4.0 minutes. In various embodiments, the controller can be configured to perfuse the culture medium for about 5 seconds to about 4 minutes, about 10 seconds to about 3.5 minutes, about 15 seconds to about 3 minutes, about 15 seconds to about 2 minutes, about 25 seconds seconds to about 90 seconds, about 30 seconds to about 75 seconds, about 40 seconds to about 2.0 minutes, about 60 seconds to about 2.5 minutes, about 90 seconds to about 3.0 minutes, or 1.8 minutes to about 4 minutes. The flow controller (not shown) can be configured to perfuse the first fluid medium into the flow region at a rate that is much greater than the average rate at which the components diffuse from the isolated region of the growth chamber into the flow channel. For example, the fluid flow rate in the flow region may be about 0.009 microliters/second, about 0.01 microliters/second, about 0.02 microliters/second, about 0.03 microliters/second, about 0.05 microliters/second, about 0.1 microliters /sec, about 0.2 μl/sec, about 0.3 μl/sec, about 0.4 μl/sec, about 0.5 μl/sec, about 0.6 μl/sec, about 0.7 μl/sec, about 0.8 μl/sec sec, about 0.9 µl/sec, about 1.0 µl/sec, about 1.1 µl/sec, about 1.2 µl/sec, about 1.3 µl/sec, about 1.4 µl/sec, about 1.5 µl/sec , about 1.7 µl/sec, about 1.9 µl/sec, about 2.0 µl/sec, about 2.2 µl/sec, about 2.4 µl/sec, about 2.6 µl/sec, about 2.8 µl/sec, about 3.0 µl/sec, about 3.2 µl/sec, about 3.4 µl/sec, about 3.6 µl/sec, about 3.8 µl/sec, about 4.0 µl/sec, about 4.2 µl/sec, about 4.4 microliters/sec, about 4.6 microliters/sec, about 4.8 microliters/sec, about 5.0 microliters/sec, about 6.0 microliters/sec, about 7.0 microliters/sec, about 8.0 microliters/sec, or about 9.0 μl/sec, any of which is a rate that will sweep a connecting region of the growth chamber (but not an isolated region of the growth chamber(s)). The controller can provide a velocity of the first fluid medium which is a non-sweep rate of the velocity of the fluid medium, i.e., below V _max(Maximum velocity of the microfluidic device that avoids rupture of the microfluidic device due to excessive pressure and restricts the movement of components between a second fluid medium in the growth chamber and a first fluid medium in the flow region to the diffusion region ) at any suitable rate. In some embodiments, the controller can be configured to perfuse the first fluid medium through the flow area at about 0.05 microliters/second, about 0.06 microliters/second, about 0.07 microliters/second, about 0.08 microliters/second µl/sec, about 0.09 µl/sec, about 0.10 µl/sec, about 0.11 µl/sec, about 0.12 µl/sec, about 0.13 µl/sec, about 0.14 µl/sec, about 0.15 µl liter/sec, about 0.16 microliter/sec, about 0.17 microliter/sec, about 0.18 microliter/sec, about 0.19 microliter/sec, about 0.20 microliter/sec, about 0.30 microliter/sec, about 0.40 microliter /sec, about 0.50 μl/sec, about 0.60 μl/sec, about 0.70 μl/sec, about 0.80 μl/sec, about 0.90 μl/sec, about 1.00 μl/sec, about 1.10 μl/sec sec, about 1.20 microliters/sec, about 1.30 microliters/sec, about 1.40 microliters/sec, about 1.50 microliters/sec, about 1.60 microliters/sec, about 1.70 microliters/sec, about 1.80 microliters/sec , about 1.90 µl/sec, about 2.00 µl/sec, about 2.10 µl/sec, about 2.20 µl/sec, about 2.30 µl/sec, about 2.40 µl/sec, about 2.50 µl/sec, About 2.60 microliters/second, about 2.70 microliters/second, about 2.80 microliters/second, about 2.90 microliters/second, or about 3.00 microliters/second. In some embodiments, the controller can be configured to perfuse the first fluid medium through each of the plurality of flow regions according to: about 0.01 microliters/second, about 0.02 microliters/second, about 0.03 microliters /sec, about 0.04 μl/sec, about 0.05 μl/sec, about 0.06 μl/sec, about 0.07 μl/sec, about 0.08 μl/sec, about 0.09 μl/sec, about 0.10 μl/sec seconds or about 0.11 µl/sec. In various embodiments, the flow rate and duration of infusion provide at least about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 10, about 11, about 15, about 20, about 25, about 30, about 35, about 50, about 75, about 100, about 200, about 300, or more than 300 volumes The total amount of a fluid medium. In various embodiments, perfusion may be accomplished using varying durations, varying flow rates, and perfusion stop durations, as shown in the methods of FIGS. 7 and 8 and discussed below. Reservoirs, Media Conditioning and Introducing Groups MinuteThe system can further include a reservoir configured to contain a fluid medium that can be introduced at the inlet 124 of the microfluidic device and can be perfused by a flow controller. The reservoir can be fluidly connected at an upstream location to any of the microfluidic devices as described herein (non-limiting examples include 100, 200, 240, 290, or 400) ( picture 5A to figure 5E). The fluid medium can be adjusted in the reservoir to contain the desired gas balance (i.e., for a non-limiting example, a mixture containing 5% carbon dioxide) that provides optimal growth for the cells being cultured, and also moderates micro-organisms. pH in fluidic devices. In some embodiments, the reservoir may further contain a cell population different from the cells of interest in the microfluidic device. This population of cells can be feeder cells that produce soluble signaling or growth factors required for cell growth and/or survival in the microfluidic device. In this way, the fluid medium can be adjusted for optimal growth and/or viability prior to introduction into the microfluidic device. Using a reservoir to house the feeder cell population prevents contamination of the cultured cell population in the microfluidic device; soluble secretions from the feeder cells can be incorporated into the fluid medium delivered into the microfluidic device, but fed Cells may not be aspirated upwards with the fluid medium. An embodiment of a reservoir, conditioning, and introduction components of a system is shown in FIG. 5A. In this embodiment, the reservoir may be another microfluidic device 502 containing a fluid medium 202 (not shown) conditioned within the microfluidic device 502. The microfluidic device 502 has an enclosure 510 and a substrate 512, at least one of which is breathable. Microfluidic device 502 may also contain a population of feeder cells maintained such that the feeder cells produce soluble growth factors or other cell signaling components required for cell growth and/or survival in microfluidic device 500A. The reservoir 502 can be housed in a chamber 516 to provide a 5% carbon dioxide gas environment, such as (but not limited to) a gas environment. The fluid medium 202 in the reservoir 502 absorbs the gas mixture (eg, 5% carbon dioxide in air) passing through the gas permeable walls of the reservoir, and also absorbs soluble secretions from the feeding cells. The culture medium 202 is perfused into the microfluidic device 500A via the inlet 124 from the reservoir 502 by the pump 514, through the gas-impermeable connecting conduit 506, and forms a flow 212 in the flow channel 134 of the microfluidic device 500A. In this embodiment, the pump connection conduit 504 (not labeled), the transfer connection conduit 506, the substrate 104 or the closure body 102 are all gas impermeable. Fluid medium flow 212 sweeps through the growth chamber of microfluidic device 500A and allows waste components of fluid medium 204 to diffuse out of the growth chamber (not shown) while allowing components to diffuse from fluid medium 202 in flow channel 134 to in the growth room. Finally, spent fluid medium 202' (not shown) exits microfluidic device 500A via outlet 124' in outlet connection conduit 508. In another embodiment, the fluid medium 202 is transferred into the microfluidic device 500B via the pump connecting conduit 504 and through the gas permeable block 518, as shown in Figure 5B. The gas permeable block 518 is incorporated into the upper surface of the closure body 102 and forms a portion of the upper surface of the closure body 102 . Portions of the upper surface of the enclosure 102 formed by the gas permeable block 518 may be positioned above the growth chamber of the microfluidic device 500B. Microfluidic device 500B is housed within a chamber 516 that provides a gaseous environment (eg, 5% carbon dioxide) that is exchanged into the fluid medium in microfluidic device 500B. Additionally, chamber 516 may provide temperature and/or humidity regulation to microfluidic device 500B. Pump connection conduit 504, enclosure 102 or substrate 104 are all impermeable, and exchange through gas permeable block 518 can act as the "lungs" of microfluidic device 500B and properly condition the culture medium within microfluidic device 500B. In this embodiment, the fluid medium 202 may be additionally conditioned in another component prior to loading into the pump 514, and may thus also contain secreted material, eg, from a feeding cell culture. In another embodiment, the gas permeable block is integrated into the upper surface of the enclosure 102 of the microfluidic device 500C to form a gas permeable section 518', as shown in FIG. 5C. The fluid medium can be conditioned and introduced as discussed above for the embodiment of Figure 5B and can further comprise secreted material from a population of feeding cells. The microfluidic device 500C may be housed in a chamber 516 containing a gaseous environment (eg, 5% carbon dioxide in air). The gaseous environment can be exchanged across the gas permeable section 518 ′, which can be one section or a plurality of sections in the upper surface of the enclosure 102 . The chamber 516 can further appropriately adjust the temperature and humidity of the device 500C. In this embodiment, the pump connection conduit 504, the closure body 102 (except for the gas permeable block 518') and the substrate 104 may be impermeable. In some embodiments, at least one gas permeable section 518' is positioned over a growth chamber of the microfluidic device 500C. In another embodiment, at least one gas permeable section 518' is positioned over the flow region 134 of the microfluidic device 500C. In other embodiments, the gas permeable section 518 ′ may be positioned over both the at least one growth chamber and the at least one flow region 134 . In a further embodiment, the gas permeable conduit 504' can be used to condition (eg, equilibrate) the fluid medium prior to introducing the medium into the microfluidic device 500D, as shown in Figure 5D. The length of the gas permeable conduit 504' may be selected to provide sufficient surface area to allow efficient gas exchange and equilibrium within an enclosure 516, which may contain a gaseous environment such as, but not limited to, 5% carbon dioxide in air. The environment of 516 can further regulate the temperature and/or humidity of the culture medium within the gas permeable pump connection conduit 504'. A non-limiting example of a gas permeable material that can be used in a gas permeable connecting conduit is Teflon® AF. The fluid medium can be conditioned by contact with a population of feeding cells prior to introduction into the pump assembly 514 and thus can contain secretions that can optimize the growth and/or viability of the cultured cell(s) in the microfluidic device 500D substance. The previous conditioning using the feeder cell population can take place within chamber 516 or can be performed in another culture component that itself has environmental control over any of temperature, humidity, pH and/or gas environment. In this embodiment, the enclosure 102 and substrate 104 of the microfluidic device 500D may be gas impermeable. In yet another embodiment of the system's reservoir, media conditioning, and introduction components, the media may be conditioned in a reservoir 502' that can be placed in a suitable gaseous environment, as shown in Figure 5E. The reservoir 502' need not be a microfluidic device or any particular type of culture component. The reservoir 502' is placed in a suitable gaseous environment such as, for example, 5% carbon dioxide in air by providing a connected supply 526 from a gaseous environment source 524. The fluid medium within reservoir 502' has gas exchange with the gaseous environment provided from source 524, and is conditioned thereby. The fluid medium in reservoir 502' may also contain a medium that feeds the cells to provide secreted material that can optimize the growth and/or viability of cultured cells in microfluidic device 500E. Conditioning fluid medium can be transferred from reservoir 502' via transfer connection conduit 522 (which is connected to a valve 520 on a pump 514) and can be injected into channel 134 of microfluidic device 500E by pump 514 via connection conduit 504 . The fluid medium injected into the microfluidic device 500E forms fluid flow 212 . After passing through flow channel 134, spent fluid medium 202' exits microfluidic device 500E via outlet conduit 508. In this embodiment, the transfer connection conduit 522, the connection conduit 504, the valve 520, the pump 514, the closure body 102, and the substrate 104 may all be gas impermeable. In some embodiments, the connecting conduit 526 connecting the source 524 to the reservoir 502' may be substantially gas impermeable. In other embodiments, the connecting conduit 526 need not be substantially gas impermeable, but may be relatively gas impermeable. In some embodiments shown in Figure 5E, the gas (not shown) may be continuously flowing or may be pulsed, eg, the periodic replacement gas input from source 524 (not shown) may be air 5% carbon dioxide. In other embodiments, the gas input from source 524 may be 100% carbon dioxide. When 100% carbon dioxide gas is used, a small amount of carbon dioxide gas may be injected into the headspace (not shown) of the reservoir 502' to maintain a 5% carbon dioxide mixture in the headspace. In some embodiments, when the gas is injected into the headspace of the reservoir 502', the reservoir 502' may further include a fan (not shown) to keep the injected gas from the headspace (not shown) not shown) are mixed with other gas components (not shown). In some embodiments, the lid 102 of the microfluidic device 500E may have a carbon dioxide sensor (not shown) incorporated or attached therein when the gas input is pulsed. In some embodiments, 100% carbon dioxide gas may be input from source 524 for cost savings over using a commercial 5% carbon dioxide gas mixture in air. In other embodiments, 100% carbon dioxide gas may be introduced into source 524 and mixed with air to produce a 5% carbon dioxide mixture in air. In any of the above-described embodiments, the chamber 516 may be further humidified such that the gaseous environment of the chamber does not alter the osmotic pressure of the microfluidic device and/or the fluid medium in the reservoir. In another embodiment, an alternative method of providing appropriate gas exchange for cells cultured in a growth chamber may provide gas flow through the flow region of a microfluidic device (not shown). A suitable gas (eg, 5% carbon dioxide) can be pumped directly or pulsed through the flow channel. Because the isolated regions of the growth chamber are designed to be largely unswept in volume, cells positioned within the isolated regions are not disturbed by air or air bubbles moving through the flow channels (swept regions). This will provide very fast gas exchange between the gas in the flow channel and the fluid medium in the growth chamber because the diffusion distance is very small compared to, for example, in a 50 mL conical tube. Then, the gas can be replaced by the fluid medium after any selected amount of time. The gas flow can be repeated at any desired frequency to maintain a steady concentration of dissolved gas components, which also affects the pH of the fluid medium. Alternatively, non-optimal gas compositions or repetitions can be used to perturb the environment of the cells. In summary, there are various components and configurations that can be used to provide conditioned medium to cells in the growth chambers of the microfluidic devices described herein. Any of the

microfluidic devices

100, 200, 240, 290, or 400 may be used with any of the embodiments of Figures 5A-5E. The systems and kits can include connecting conduits configured to connect to the inlet and/or outlet of the microfluidic device. The connecting conduit may also be configured to connect to the reservoir and/or pump assembly. Accordingly, there is provided a microfluidic device for culturing one or more biological cells comprising: a flow region configured to contain a flow of a first fluid medium; and at least one growth chamber comprising a flow through at least one surface conditioned to support cell growth, viability, transplantability, or any combination thereof within the microfluidic device, wherein the at least one growth chamber comprises an isolation region and a connection region, the isolation region and the connection The regions are fluidly connected and the connecting region includes a proximal opening to the flow region. In various embodiments, the isolation region of the microfluidic device can be configured to contain a second fluidic medium. When the flow region and the at least one growth chamber are substantially filled with the first fluid medium and the second fluid medium, respectively, components of the second fluid medium can diffuse into the first fluid medium and/or the second fluid medium Components of a fluid medium can diffuse into the second fluid medium; and the first fluid medium does not flow substantially into the isolation region. In various embodiments, the at least one conditioning surface can be conditioned to support the transplantability of the one or more biological cells within the microfluidic device. In some embodiments, portions of the conditioning surface can be configured to support the transplantability of the biological cells within the microfluidic device. In some embodiments, the at least one conditioning surface of the microfluidic device can comprise a polymer comprising an alkylene ether moiety. In other embodiments, the at least one conditioning surface of the microfluidic device may comprise a polymer comprising a carboxylic acid moiety, a sulfonic acid moiety, a nucleic acid moiety or a phosphonic acid moiety. In other embodiments, the at least one conditioning surface of the microfluidic device may comprise a polymer comprising a carbohydrate moiety. In some embodiments, the polymer comprising saccharide moieties may be polydextrose. In some other embodiments, the at least one conditioning surface of the microfluidic device can comprise a polymer that includes amino acid moieties. Alternatively, the at least one conditioning surface of the microfluidic device may comprise one or more components of mammalian serum. These components of mammalian serum can be supplements to a culture medium. In some embodiments, the mammalian serum can be fetal bovine serum (FBS) or calf serum (FCS). In various embodiments of the microfluidic device, the at least one conditioning surface may comprise carbohydrate moieties. In some embodiments, the at least one conditioning surface may comprise alkylene ether moieties. In other embodiments, the at least one conditioning surface may comprise amino acid moieties. In some other embodiments, the at least one conditioning surface may comprise alkyl or perfluoroalkyl moieties. In some embodiments, the alkyl or perfluoroalkyl moieties can have a backbone length greater than one of 10 carbons. In some embodiments, the at least one conditioning surface may comprise a moiety, which may be: an alkyl or fluoroalkyl (which includes perfluoroalkyl) moieties; a monosaccharide or polysaccharide (which may include, but is not limited to) polydextrose ); alcohols (including but not limited to propargyl alcohol); polyols, including but not limited to polyvinyl alcohol; alkylene ethers, including but not limited to polyethylene glycol; polyelectrolytes (which includes, but is not limited to, polyacrylic acid or polyvinylphosphonic acid); amine groups (which include derivatives thereof, such as, but not limited to, alkylated amine groups, hydroxyalkylated amine groups, guanidine salts, and those containing a non- Heterocyclyl groups that aromatic nitrogen ring atoms, such as, but not limited to, morpholinyl or piperazinyl); carboxylic acids, including but not limited to propyonic acid (which can provide a carboxylate anion surface); phosphonic acid , which include, but are not limited to, ethynylphosphonic acid (which can provide a phosphonic acid anion surface); sulfonic acid anion; carboxybetaine; sulfobetaine; amine sulfonic acid; or amino acid. In various embodiments of the microfluidic device, the at least one conditioning surface can include a linking group covalently bonded to a surface of the microfluidic device, and the linking group can be bound to the conformed to support cell growth, viability, transplantability or any combination thereof within the microfluidic device. In some embodiments, the linking group can be a siloxy linking group. In other embodiments, the linking group can be a phosphonate linking group. In some embodiments, the linking group can be directly linked to the moiety that is configured to support cell growth, viability, transplantability, or any combination thereof. In some embodiments, the portion of the conditioning surface can be configured to support the transplantability of the biological cells within the microfluidic device. In other embodiments, the linking group can be directly linked to the moiety that is configured to support cell growth, viability, transplantability, or any combination thereof. In other embodiments, the linking group may be indirectly linked via a linker to the moiety configured to support cell growth, viability, transplantability, or any combination thereof. In various embodiments, the linker may comprise a triazolyl moiety. In various embodiments of the microfluidic device, the at least one conditioning surface may comprise zwitterions. In other embodiments, the at least one conditioning surface may comprise phosphonic acid moieties or carboxylic acid moieties. In other embodiments, the conditioning surface may contain anions. In some other embodiments, the at least one conditioning surface may comprise amine groups or guanidine moieties. In other embodiments, the at least one conditioning surface may comprise cations. In various embodiments of the microfluidic device, the at least one modulating surface can comprise at least one cell adhesion blocking molecule. The at least one cell adhesion blocking molecule can disrupt actin fibril formation, block integrin receptors, or reduce cell binding to DNA-fouled surfaces. The at least one cell adhesion blocking molecule can be cytochalasin B, an RGD-containing peptide or a DNase 1 protein. In other embodiments, the at least one cell adhesion blocking molecule may comprise a combination of more than one type of cell adhesion blocking molecule. In various embodiments of the microfluidic device, the at least one conditioning surface is configured to be heated to a temperature of at least about 30°C. The at least one conditioning surface can be configured to be stable at a temperature of at least about 30°C. In various embodiments of the microfluidic device, the microfluidic device can further comprise a microfluidic channel comprising at least a portion of the flow region. In some embodiments, the connection region of the at least one growth chamber may directly open into the microfluidic channel. In some embodiments, the isolation region of the at least one growth chamber of the microfluidic device may have dimensions sufficient to support cell expansion to a range of about 100 cells. In some embodiments, no more than 1×10 can be maintained in the at least one growth chamber ²biological cells. In some embodiments, the volume of the at least one growth chamber can be less than or equal to about 2×10 ⁶cubic microns. In other embodiments, no more than 1 x 10 may be maintained in the at least one growth chamber ²biological cells, and the volume of the at least one growth chamber may be less than or equal to about 1×10 ⁷cubic microns. In various embodiments of the microfluidic device, the microfluidic device can further comprise at least one inlet configured to input the first fluidic medium or the second fluidic medium into the flow region and configured to The first medium receives at least one outlet of the first medium as it exits the flow region. In various embodiments of the microfluidic device, the microfluidic device can further comprise a substrate having a dielectrophoretic (DEP) configuration configured to introduce one or more biological cells into the growth chamber or to The one or more biological cells are removed from the growth chamber. The DEP configuration can be actuated optically. In various embodiments of the microfluidic device, the microfluidic device may further comprise a deformable cover region over the at least one growth chamber or the isolation region thereof, whereby pressing the deformable cover region applies sufficient to cause the A force that biological cells output from the isolation region to the flow region. In some embodiments, the microfluidic device can include a lid, wherein at least a portion of the lid is gas permeable, thereby providing a source of gas molecules to a fluid medium positioned in the microfluidic device. In some embodiments, the gas permeable portion of the cover may be positioned over the at least one growth chamber. In some embodiments, the breathable portion of the cover may be positioned over the flow area. In some embodiments, the microfluidic device can further comprise a deformable cover region over the at least one growth chamber or the isolation region thereof, whereby pressing the deformable cover region is applied sufficient to free the biological cells from the isolation The area outputs a force to this flow area. In various embodiments of the microfluidic device, the conditioning surface may comprise a cleavable moiety. The lysable moiety can be configured to allow disruption of the regulatory surface, thereby promoting the transplantability of the one or more biological cells after culture. In various embodiments of the microfluidic device, the at least one growth chamber may comprise a plurality of growth chambers. In various embodiments of the microfluidic device, the one or more biological cells may comprise a plurality of biological cells. In some embodiments, the one or more biological cells may comprise one or more mammalian cells. In some embodiments, the one or more biological cells may comprise one or more fusion tumor cells. In some embodiments, the one or more biological cells may comprise one or more lymphocytes or white blood cells. In other embodiments, the one or more biological cells may comprise a B cell, a T cell, a NK cell, a macrophage, or a combination thereof. In various embodiments, the one or more biological cells can comprise one or more adherent cells. In some embodiments, the one or more biological cells in the growth chamber can be a single cell and the colony can be a colony of biological cells. pH sensorThe system may further include at least one sensor connected to at least one inlet 124 and/or at least one outlet 124' of microfluidic device 600 (as shown in FIG. 6). Alternatively, device 600 may be any of

devices

100, 200, 240, 290, 400, or 500A-500E. The sensor can be configured to detect a pH of the first fluid medium as it enters the microfluidic device 600 . Alternatively, the sensor may be configured to detect a pH of the first fluid medium as it exits the microfluidic device 600 . The sensor may be incorporated into the microfluidic device or it may be capable of being attached to or in line with an inlet 124 and/or an outlet 124' of the microfluidic device Separate components one by one in a row. In some embodiments, the pH sensor is an optical sensor. An optical sensor can provide an advantage over electrode-based benchtop devices because electrode-based devices can include bulky probes to make pH measurement of small (microliter) volumes difficult or impossible. Similarly, in-line flow-through solutions can have very long settling times (5 minutes to 15 minutes) due to the nature of the microelectrodes, and epitaxy calibration procedures may be required before each use. Furthermore, electrodes can deteriorate rapidly and thus require more maintenance. The optical sensor may be an integrated electrodeless device including an LED for illumination and an integrated colorimetric sensor for visible color detection. The colorimetric sensor can be a color-sensitive phototransistor. The colorimetric sensor can detect in the visible wavelength region (eg, about 390 nm to about 700 nm). Media stained with a pH-dependent dye such as, but not limited to, phenol red, can provide an instantaneous and contactless optical signal. An optical electrodeless measurement method using this optical sensor requires no contact with the culture medium and no calibration on the part of the user. Optical measurements can be calibrated to eliminate temperature dependencies. Additionally, the use of an optical sensor minimizes the risk of contaminating the sensor and thus reduces maintenance or replacement. The miniaturization of the light source (LED) and color sensor also makes this amenable to testing of very small volumes of liquids (<1 microliter) and integration into portable or hand-held instruments. The system may include drive electronics for the control/monitoring device 180 via LEDs and phototransistor sensors, and may further provide an alarm component by the control module 172 when detection of pH determines that the pH is outside the desired range. Additionally, since the settling time for color detection is fast (sub-second), this sensor can be inserted in a feedback loop to adjust the pH of the medium by modulating the carbon dioxide content in a gaseous environment surrounding the medium. Alternatively, the control module 172 or the control/monitoring device 180 may further provide components to modulate the pH of the introduced fluid medium by adding buffers and/or acidic or basic medium components to correct the pH back to the desired range. In some embodiments, sensor 610 is connected to fluidic medium inlet conduit 606 proximate to at least one inlet 124 of the microfluidic device. Conduit 606 may be transparent, substantially transparent, or translucent. The LED 614 illuminates the tube 606 and the dyed fluid medium 202a' within the tube 606. An integrated colorimetric sensor 612 can monitor the pH of the incoming fluid medium, determine that the pH has a value within a desired range for a particular culture experiment, and alarm when the pH is outside the desired range. In some embodiments, sensor 610' is connected to fluid medium outlet conduit 608, proximate to at least one outlet 124' of the microfluidic device. Conduit 608 may be transparent, substantially transparent, or translucent. The LED 614' illuminates the tube 608 and the dyed effluent fluid medium 202a'' within the tube 608. An integrated colorimetric sensor 612' can monitor the pH of the incoming fluid medium, determine that the pH has a value within a desired range for a particular culture experiment, and alarm when the pH is outside the desired range. cellA cell that can be used in the systems and methods of the present invention can be any type of cell. For example, the cell can be an embryonic cell, an oocyte or sperm cell, a stem cell, a precursor cell or a cell dissociated from a tissue, a blood cell, a fusion tumor cell, a cultured cell, a cell from a cell line, a cell cancer cells, an infected cell, a transfected and/or transformed cell (a cell line including, but not limited to, Chinese Hamster Ovary (CHO) cells), a reporter cell, or the like. The cell can be a A mammalian cell or cell can be a non-mammalian cell. The cell can comprise a bacterium, a fungus, a protozoan, or a mammalian cell infected with a parasitic species (eg, Leishmania or Plasmodium falciparum). In some embodiments, the mammalian cells can be human, murine, porcine, or any other mammal of interest. In some embodiments, cells can be from a population of cells actively growing in culture or obtained from a fresh tissue sample (eg, by dissociation of a solid tissue sample such as a biopsy or fine needle aspiration), blood , saliva, urine or other body fluids. Alternatively, the one or more biological cells can be derived from a culture medium of another sample previously frozen. In some embodiments, the one or more biological cells may comprise one or more fusion tumor cells. In other embodiments, the one or more biological cells may comprise one or more lymphocytes or white blood cells. In some embodiments, the cell line is a B cell, a T cell, a NK cell, a dendritic cell, a macrophage or other immune cell type or a precursor thereof, such as a precursor cell or a hematopoietic stem cell. In various embodiments, the one or more biological cells are one or more adherent cells. When one or more adherent cells are introduced into the microfluidic device, additional conditioning treatments can be provided to provide the adherent cells with appropriate soluble or insoluble environmental factors (eg, one or more extracellular stromal components) to Persistent survival and/or cell proliferation is allowed. Depending on the specific goals of the experiment, only one cell or a plurality of cells can be introduced into the microfluidic device for cultivation and/or colonization. When only one cell is introduced into one of the growth chambers of the system and grown according to the methods described herein, the resulting expanded population system is initially introduced into one of the colonies of the cells in the growth chamber. methodThe present invention provides a method for culturing at least one biological cell in a system comprising a microfluidic device having at least one growth chamber and a flow region. Cultivating a cell (or cells) in a growth chamber of a microfluidic device that also has a flow area may allow for the specific introduction of nutrients, growth factors or other cell signaling species at selected time periods to achieve cell growth, viability or availability Control of portability parameters. The at least one biological cell is introduced into the at least one growth chamber having at least one regulated surface, wherein the regulated surface supports cell growth, viability, transplantability, or any combination thereof. In some embodiments, the conditioned surface supports cell transplantability within the microfluidic device. In some embodiments, transplantability includes preventing cells from adhering to the microfluidic device. In other embodiments, transplantability comprises providing adherent cells with a conditioned surface that will support cell growth, viability, transplantability, or any combination thereof, while also allowing for the microfluidic device to be used after a culture cycle mobile cells. The at least one conditioning surface can be any conditioning surface as described herein. Introduction of the at least one biological cell can be accomplished using several different dynamics as described herein, portions of which can allow for placement of a particular biological cell into a particular location on the microfluidic device (eg, into a preselected growth chamber) for precise control. The precise control of cell placement/removal and nutrient/signaling/environmental stimuli that can be accomplished by the methods described herein is difficult or impossible to achieve using large-scale or other microfluidic culture methods. Following placement, the at least one biological cell is incubated for a period of time at least sufficient to allow the at least one biological cell to expand to produce a colony of biological cells. When biological cells are introduced into a separate growth chamber, the resulting expanded population can be precisely identified for further use as a separable population of biological cells. When only one biological cell is introduced into a growth chamber and the biological cell is allowed to expand, the resulting colony is a colony of one of the biological cells. Any suitable cells, including but not limited to cells as described above, can be used in these methods. The microfluidic device can be any of the

microfluidic devices

100, 300, 400, 500A to 500E or 600 as described herein, and the microfluidic device can be any one of having components as described herein part of the system. The at least one growth chamber can include a plurality of growth chambers, and any suitable number of growth chambers as discussed herein can be used. In some embodiments of the methods, the microfluidic device can have about 500 to about 1500 growth chambers, about 1000 to about 2000 growth chambers, about 1000 to about 3500 growth chambers, about 2000 to about 2000 growth chambers about 5000 growth chambers, about 3000 to about 7000 growth chambers, about 5000 to about 10000 growth chambers, about 7500 to about 15000 growth chambers, about 10000 to about 17500 growth chambers, or about 12500 to about 12500 growth chambers About 20,000 growth chambers. In the method of culturing one or more biological cells, the at least one regulated surface can be any regulated surface as described herein. The conditioning surface can be covalently bonded to the microfluidic device. In some embodiments, the conditioning surface can include a linking group covalently bonded to the surface, and the linking group can also be linked to the one or the other configured to support the microfluidic device. A portion of cell growth, viability, transplantability, or any combination thereof, of a plurality of biological cells. In some embodiments, a microfluidic device with a conditioning surface may be provided prior to infusion of the one or more biological cells. Introduce at least one biological cellIn some embodiments, introducing the at least one biological cell into the at least one growth chamber can include using a dielectrophoretic (DEP) force having a force sufficient to move the at least one biological cell. The DEP force can be generated using electrical tweezers, such as optoelectronic tweezers (OET). In some other embodiments, introducing one or more biological cells into the at least one growth chamber can include: using fluid flow and/or gravity (eg, by tilting the microfluidic device) to cause the cell(s) Drop into a growth chamber positioned below the cell(s). In some embodiments, the at least one biological cell is introduced into the microfluidic device through an inlet 124 in a flow region (eg, flow channel) of the microfluidic device. The flow of medium in the flow channel can transport the cells to a location proximate an opening of a growth chamber. After the biological cell is positioned proximate an opening in a growth chamber, the biological cell can be moved into the growth chamber using any of the forces described herein, including dielectrophoresis or gravity. Dielectrophoretic forces may include electrical or optical actuation forces, and these DEP forces may further be provided by optoelectronic tweezers (OET). The at least one biological cell is movable through the flow channel to the proximal opening of a connecting region of the at least one growth chamber, wherein the connecting region opens directly to the flow channel/region and is fluidly connected to the flow channel/region. The connection region of the at least one growth chamber is also fluidly connected to an isolation region of the at least one growth chamber. The at least one biological cell can move further through the connecting region into the isolation region of the at least one growth chamber. The isolation region of the at least one growth chamber can be of sufficient size to support cell expansion. Typically, however, the size of the growth chamber will limit this expansion to no more than about 1 x 10 ³pcs, about 5×10 ²pcs, about 4×10 ²pcs, about 3×10 ²pcs, about 2×10 ²pcs, about 1×10 ²individual, about 50, about 25, about 15, or even only 10 cultured cells. In some embodiments, the isolation region may be of sufficient size to support cell expansion to no more than about 1 x 10 ²A size of about 50, about 25, about 15 or about 10 cultured cells. We have surprisingly found that up to about 1 x 10 can be successfully performed in an isolated region having a volume of no more than one of ²Cell culture and/or expansion of individual cells: about 1.0×10 ⁷Cubic microns, about 6×10 ⁶Cubic microns, about 2×10 ⁶Cubic micron, about 1.5×10 ⁶cubic microns or about 1.0 x 10 ⁶cubic microns. In some other embodiments, there may be no more than about 4 x 10 ⁵Cubic microns successfully performed up to about 1 x 10 in an isolated region of one volume ²Cell culture and/or expansion of individual cells. The size of the biological cells can vary widely from the following depending on the cell type: bacteria having a diameter of about 1 micrometer and a volume of about 1 cubic micrometer, bacteria having a diameter of about 7 micrometers to about 8 micrometers, and a diameter of about 100 micrometers A small human cell (such as a red blood cell) in a volume of cubic micrometers, an immortalized cell line (such as HeLa) with a diameter of about 40 micrometers (non-confluent) and a volume of about 2000 cubic micrometers ), a megakaryocyte having a diameter of about 25 microns up to about 60 microns and a volume of about 4700 cubic microns to about 100,000 cubic microns, or a megakaryocyte having a diameter of about 120 microns and a volume of about 900,000 cubic microns human egg cells. According to this, with about 4×10 ⁵A growth chamber with a volume of cubic micrometers can accommodate larger species of megakaryocytes (approximately 1 x 10 ⁵volume in cubic microns) expanded very little, eg, less than 5 cells in total. Alternatively, the same small growth chamber (approximately 4 x 10 ⁵cubic micron volume) allows bacterial cells (about 1 cubic micron volume) to expand to about 400,000 bacterial cells. The method may further comprise: introducing a first fluidic medium into a microfluidic channel of the flow region of the microfluidic device. In some embodiments, introducing the first fluid medium is performed prior to introducing the at least one biological cell. When introducing the first fluidic medium prior to introducing the at least one biological cell, a flow rate can be selected such that the first fluidic medium, for example, flows from the flow channel of the microfluidic device to the growth chamber at any suitable rate middle. Alternatively, if the microfluidic device has been pre-filled with a medium containing too much one or more conditioning reagents, the first fluidic medium flows into the microfluidic channel at a rate such that the first fluidic medium replaces the Any remaining medium in the flow area that contains too much conditioning reagent(s). When the flow of the first fluid medium is introduced after the at least one biological cell is introduced into the growth chamber, the flow rate of the first fluid medium can be selected so as not to sweep the isolation region that would cause the at least one The biological cells are displaced from the isolated area. The fluid medium surrounding the at least one biological cell in the isolated region of the at least one growth chamber is a second fluid medium, which may be the same as or different from the first fluid medium. In some embodiments, the second fluid medium can be the same as the first fluid medium, but during the incubation step, cell waste and depleted medium components can cause the second fluid medium to behave differently from the first fluid culture medium. growing cellsIn the methods described herein, cells are grown for a period of time at least sufficient to expand at least one biological cell to produce a colony of biological cells. The time period can be selected to be from about 1 day to about 10 days. In other embodiments, the incubation period can be extended beyond 10 days and can be extended for any desired period. Cells can grow indefinitely because cells in isolated areas of the growth chamber are supplied with nutrients and have waste removed by perfusion of fluid medium. Because the isolation area is filled with a population of expanded cells, any additional expansion will result in expanded biological cells inhabiting the junction area of the growth chamber, which is a swept area of the growth chamber. Perfusion of the medium may allow the expanded biological cells to be swept out of the junction area of the growth chamber and subsequently out of the microfluidic device. Accordingly, the number of cells present in the isolation region of the growth chamber can be stabilized at a maximum number that depends on the size of the biological cells and the size of the isolation region in the growth chamber. The ability to stabilize the maximum number of cells in an isolated population of cells provides one advantage over other currently available methods of cell culture because lengthy cell population divisions can be eliminated. In some embodiments, incubation can be performed for about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 7 days, about 8 days, about 9 days, about 10 days, or more than 10 days. The incubation period can range from about 1 day to about 6 days, from about 1 day to about 5 days, from about 1 day to about 4 days, from about 1 day to about 3 days, or from about 1 day to about 1 day 2 days. In other embodiments, incubation can be performed for less than about 5 days, less than about 4 days, less than about 3 days, or less than about 2 days. In some embodiments, incubation can be performed for less than about 3 days or less than about 2 days. In other embodiments, incubation can be performed for about 3 hours, about 4 hours, about 5 hours, about 6 hours, about 7 hours, about 8 hours, about 9 hours, about 10 hours, about 11 hours, about 12 hours, About 13 hours, about 14 hours, about 15 hours, about 16 hours, about 17 hours, about 18 hours, about 19 hours, about 20 hours, about 21 hours, about 22 hours, or about 23 hours. During the culturing step, an image of the at least one growth chamber and any cells contained therein can be monitored at one or more time points throughout the culturing step. The image may be stored in memory in a processing component of the system. perfused cellsDuring the incubation step, the second fluid medium present in the isolated region of the growth chamber may become depleted of nutrients, growth factors or other growth stimulators. The second fluid medium can accumulate cellular waste. Additionally, because at least one biological cell continues to grow during the incubation cycle, it may be desirable to change nutrients, growth factors or other growth stimulators to different nutrients, growth factors or other growth factors from the first or second medium at the start of the incubation stimulant. As described herein, cultivation in a growth chamber of a microfluidic device can impart the specific and selective ability to introduce and modify chemical gradients sensed by at least one biological cell that can more closely approximate in vivo conditions. Alternatively, intentionally altering the chemical gradient sensed by at least one biological cell to a non-optimized set of conditions may allow cells to expand under conditions designed to explore disease or therapeutic pathways. Accordingly, the method may comprise perfusing a first fluid medium during the incubation step, wherein the first fluid medium is introduced via the at least one inlet 124 of the microfluidic device and wherein the output via the at least one outlet of the microfluidic device optionally includes from the second The first fluid medium of the components of the fluid medium. The exchange of components of the first fluid medium to thereby provide fresh nutrients, soluble growth factors and the like and/or exchange of waste components of the medium surrounding the cell(s) within the isolation region occurs substantially under diffusion conditions The interface between the swept area and the unswept area of the growth chamber. We have surprisingly found that efficient exchange occurs under substantially no-flow conditions. Accordingly, we have surprisingly discovered that continuous perfusion is not required for successful breeding. Thus, perfusion can be discontinuous. In some embodiments, the perfusion is periodic, and in some embodiments, the perfusion is irregular. Interruptions between perfusion cycles may be sufficient to allow components of the second fluid medium in the isolation region to diffuse into the first fluid medium in the flow channel/region and/or components of the first fluid medium to diffuse into the second fluid medium The first medium did not flow substantially into the isolation area for the duration of . In another embodiment, a low perfusion rate can also be used to obtain efficient exchange of the components of the fluid medium within the unswept and outside regions of the growth chamber. Accordingly, a method of perfusing at least one biological cell in at least one growth chamber of a microfluidic device is shown in FIG. 7 and includes a perfusion step 7002, wherein the first fluid medium is at a first perfusion rate R ₁flow through a flow region of the microfluidic device into a flow region fluidly connected to the growth chamber for a first perfusion time D ₁. R ₁Can be selected to be a non-sweep and flow rate, as described herein. The method 700 further includes the step 7004 of stopping the flow of the fluid medium for a first perfusion stop time S ₁.

Steps

7002 and 7004 are repeated W times (where W may be an integer selected from 1 to about 1000), after which the perfusion procedure 700 is complete. In some embodiments, W may be an integer from 2 to about 1000. Another method 800 of perfusing at least one biological cell in at least one growth chamber of a microfluidic device is shown in FIG. 8, which includes a first perfusion cycle comprising step 8002: subjecting a fluid medium to a first perfusion cycle a perfusion rate R ₁flow through a flow region of the microfluidic device into a flow region fluidly connected to the growth chamber for a first perfusion time D ₁. R ₁Can be selected to be an unswept and flow rate, as described herein. The first perfusion cycle includes step 8004: stopping the flow of the fluid medium for a first perfusion stop time S ₁. The first perfusion cycle can be repeated W times, where W is an integer selected from 1 to about 1000. After W repetitions of the first perfusion cycle are completed, the method 800 further includes a second perfusion cycle comprising step 8006: subjecting the first fluid medium to a second perfusion rate R ₂flow for a second perfusion time D ₂, where R ₂A non-sweep and flow rate is selected. The second perfusion cycle of method 800 further includes step 8008: stopping the flow of fluid medium for a second perfusion stop time S ₂. Thereafter, the method returns to

steps

8002 and 8004 of the first priming cycle and repeats the combined two-cycle priming procedure V times, where V is an integer from 1 to about 5000. The combination of W and V can be selected to meet the desired end of the incubation period. In various embodiments of

method

700 or 800, the perfusion rate R ₁It can be any non-sweep and flow rate of the fluid medium as described above for the flow controller configuration. In some embodiments, R ₁can be about 0.009 microliters/sec, about 0.010 microliters/sec, about 0.020 microliters/sec, about 0.030 microliters/sec, about 0.040 microliters/sec, about 0.05 microliters/sec, about 0.06 microliters/sec , about 0.07 µl/sec, about 0.08 µl/sec, about 0.09 µl/sec, about 0.10 µl/sec, about 0.11 µl/sec, about 0.12 µl/sec, about 0.13 µl/sec, about 0.14 µl/sec, about 0.15 µl/sec, about 0.16 µl/sec, about 0.17 µl/sec, about 0.18 µl/sec, about 0.19 µl/sec, about 0.20 µl/sec, about 0.30 µl/sec, about 0.40 µl/sec, about 0.50 µl/sec, about 0.60 µl/sec, about 0.70 µl/sec, about 0.80 µl/sec, about 0.90 µl/sec, about 1.00 µl/sec, about 1.10 µl/sec, about 1.20 µl/sec, about 1.30 µl/sec, about 1.40 µl/sec, about 1.50 µl/sec, about 1.60 µl/sec, about 1.70 µl L/sec, about 1.80 µl/sec, about 1.90 µl/sec, about 2.00 µl/sec, about 2.10 µl/sec, about 2.20 µl/sec, about 2.40 µl/sec, about 2.50 µl /sec, about 2.60 microliters/sec, about 2.70 microliters/sec, about 2.80 microliters/sec, about 2.90 microliters/sec, or about 3.00 microliters/sec. In various embodiments of method 800, the second perfusion rate R ₂It can be any non-sweep and flow rate of the fluid medium as described above for the flow controller configuration. In some embodiments, R ₂Can be 0.009µl/sec, 0.010µl/sec, 0.020µl/sec, 0.030µl/sec, 0.040µl/sec, 0.05µl/sec, 0.06µl/sec, 0.07µl/sec, 0.08 µl/sec, 0.09 µl/sec, 0.10 µl/sec, 0.11 µl/sec, 0.12 µl/sec, 0.13 µl/sec, 0.14 µl/sec, 0.15 µl/sec, 0.16 µl L/sec, 0.17 µl/sec, 0.18 µl/sec, 0.19 µl/sec, 0.20 µl/sec, 0.30 µl/sec, 0.40 µl/sec, 0.50 µl/sec, 0.60 µl/sec sec, 0.70 µl/sec, 0.80 µl/sec, 0.90 µl/sec, 1.00 µl/sec, 1.10 µl/sec, 1.20 µl/sec, 1.30 µl/sec, 1.40 µl/sec, 1.50 µl/sec, 1.60 µl/sec, 1.70 µl/sec, 1.80 µl/sec, 1.90 µl/sec, 2.00 µl/sec, 2.10 µl/sec, 2.20 µl/sec, 2.40 µl L/sec, 2.50 µl/sec, 2.60 µl/sec, 2.70 µl/sec, 2.80 µl/sec, 2.90 µl/sec or 3.00 µl/sec. Selectable flow rate R ₁and/or R ₂of any combination. Typically, the perfusion rate R ₂Can be greater than the perfusion rate R ₁, and can be R ₁about 5 times, about 10 times, about 20 times, about 30 times, about 40 times, about 50 times, about 60 times, about 70 times, about 80 times, about 90 times, about 100 times, or more than 100 times. In some embodiments, R ₂than R ₁At least 10 times faster. In other embodiments, R ₂than R ₁At least 20 times faster. In yet another embodiment, R ₂Department R ₁at least 100 times the rate. In various embodiments of

method

700 or 800, the first priming time D ₁It can be any suitable priming duration as described above for the flow controller configuration. In various embodiments, D ₁Can be about 5 seconds, about 10 seconds, about 15 seconds, about 20 seconds, about 25 seconds, about 30 seconds, about 35 seconds, about 40 seconds, about 45 seconds, about 50 seconds, about 55 seconds, about 60 seconds, about 65 seconds, about 70 seconds, about 80 seconds, about 90 seconds, about 100 seconds, about 110 seconds, about 120 seconds, about 130 seconds, about 140 seconds, about 150 seconds, about 160 seconds, about 170 seconds, or about 180 seconds second. In other embodiments, D ₁It can be a range of time, such as about 10 seconds to about 40 seconds, as described above. In some embodiments, D ₁It can be from about 30 seconds to about 75 seconds. In other embodiments, D ₁Can be about 100 seconds. In other embodiments, D ₁Can range from one of about 60 seconds to about 150 seconds. In other embodiments, D ₁can be about 20 minutes, about 30 minutes, about 40 minutes, about 50 minutes, about 60 minutes, about 80 minutes, about 90 minutes, about 110 minutes, about 120 minutes, about 140 minutes, about 160 minutes, about 180 minutes, About 200 minutes, about 220 minutes, about 240 minutes, about 250 minutes, about 260 minutes, about 270 minutes, about 290 minutes, or about 300 minutes. In some embodiments, D ₁About 40 minutes to about 180 minutes. In various embodiments of

method

700 or 800, the second priming time D ₂It can be any suitable priming duration as described above for the flow controller configuration. In various embodiments, D ₂Can be about 5 seconds, about 10 seconds, about 15 seconds, about 20 seconds, about 25 seconds, about 30 seconds, about 35 seconds, about 40 seconds, about 45 seconds, about 50 seconds, about 55 seconds, about 60 seconds, About 65 seconds, about 70 seconds, about 80 seconds, about 90 seconds, or about 100 seconds. In other embodiments, D ₂It can be a range of time, such as about 5 seconds to about 20 seconds, as described above. In other embodiments, D ₂It can be from about 30 seconds to about 70 seconds. In other embodiments, D ₂Can be about 60 seconds. In various embodiments of

method

700 or 800, the first priming time D ₁May be the same as or different from the second infusion time D ₂. optional D ₁and D ₂of any combination. In some embodiments, D ₁and/or D ₂The perfusion duration can be chosen to be shorter than the stop period S ₁and/or S ₂. In various embodiments of

method

700 or 800, the first perfusion stop time S ₁Can be selected to be any suitable time period as described above for a time interval between priming cycles of the flow controller configuration. In some embodiments, S ₁can be about 0 minutes, about 5 minutes, about 10 minutes, about 15 minutes, about 20 minutes, about 25 minutes, about 30 minutes, about 35 minutes, about 40 minutes, about 45 minutes, about 60 minutes, about 65 minutes, About 80 minutes, about 90 minutes, about 100 minutes, about 120 minutes, about 150 minutes, about 180 minutes, about 210 minutes, about 240 minutes, about 270 minutes, or about 300 minutes. In various embodiments, S ₁It can be any suitable time range as described above for the time interval between infusions of the flow controller configuration, such as about 20 minutes to about 60 minutes. In some embodiments, S ₁It can be from about 10 minutes to about 30 minutes. In other embodiments, S ₁Can be about 15 minutes. In other embodiments, S ₁It can be about 0 seconds, about 5 seconds, about 10 seconds, about 20 seconds, about 30 seconds, about 40 seconds, about 50 seconds, about 60 seconds, about 70 seconds, about 80 seconds, or about 90 seconds. In some embodiments, S ₁is about 0 seconds. In various embodiments of

method

700 or 800, the second perfusion stop time S ₂Can be selected to be any suitable time period as described above for a time interval between priming cycles of the flow controller configuration. In some embodiments, S ₂can be about 0 minutes, about 5 minutes, about 6 minutes, about 7 minutes, about 8 minutes, about 9 minutes, about 10 minutes, about 20 minutes, about 30 minutes, about 45 minutes, about 50 minutes, about 60 minutes, About 90 minutes, about 120 minutes, about 180 minutes, about 240 minutes, about 270 minutes, or about 300 minutes. In various embodiments, S ₂It can be any suitable time range as described above for the time interval between infusions of the flow controller configuration, such as about 15 minutes to about 45 minutes. In some embodiments, S ₂It can be from about 10 minutes to about 30 minutes. In other embodiments, S ₂It can be about 8 minutes or about 9 minutes. In other embodiments, S ₂about 0 minutes. In various embodiments of

method

700 or 800, the first perfusion stop time S ₁and the second perfusion stop time S ₂can be independently selected from any suitable value. S ₁May be the same as or different from S ₂. In various embodiments of method 800, the number of repetitions W may be selected to be the same as or different from the number of repetitions V. In various embodiments of

method

700 or 800, W can be about 1, about 4, about 5, about 6, about 8, about 10, about 12, about 15, about 18, about 20, about 24, about 30, About 36, about 40, about 45, or about 50. In some embodiments, W can be selected to be from about 1 to about 20. In some embodiments, W may be 1. In various embodiments of method 800, V can be about 5, about 10, about 20, about 25, about 30, about 35, about 40, about 50, about 60, about 80, about 100, about 120, about 240 , about 300, about 350, about 400, about 450, about 500, about 600, about 750, about 900, or about 1000. In some embodiments, V may be selected to be from about 10 to about 120. In other embodiments, V may be selected from about 5 to about 24. In some embodiments, V may be about 30 to about 50 or may be about 400 to about 500. In various embodiments of method 800, the number of repetitions W may be selected to be the same as or different from the number of repetitions V. In various embodiments of

method

700 or 800, a total time of the first step of perfusion (represented by step 7002/7004 or 8002/8004) is from about 1 hour to about 10 hours and W is an integer of one. In various embodiments, the total time for the first step of infusion is about 9 minutes to about 15 minutes. In various embodiments of method 800, a total time of the second step of a perfusion cycle (represented by steps 8006/8008) is about 1 minute to about 15 minutes or about 1 minute to about 20 minutes. In any of

methods

700 or 800, the perfusion method may continue for the entire incubation cycle of the biological cells, eg, about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 7 days , about 8 days, about 9 days, about 10 days or more. In another non-limiting embodiment of the method 800 of FIG. 8, the controller may be configured to perfuse the fluid medium(s) into the flow region during perfusion step 8002, which has a longer perfusion period D ₁. The controller can perfuse the fluid medium at a first rate for one of the following cycles: about 45 minutes, about 60 minutes, about 75 minutes, about 90 minutes, about 105 minutes, about 120 minutes, about 2.25 hours, about 2.5 hours, about 2.45 hours, about 3.0 hours, about 3.25 hours, about 3.5 hours, about 3.75 hours, about 4.0 hours, about 4.25 hours, about 4.5 hours, about 4.75 hours, about 5 hours, or about 6 hours. In the first perfusion cycle D ₁At the end, the flow of the fluid medium can be stopped for a stop time period S ₁, S ₁It can be about 0 seconds, about 15 seconds, about 30 seconds, about 45 seconds, about 1 minute, about 1.25 minutes, about 1.5 minutes, about 2.0 minutes, about 3.0 minutes, about 4 minutes, about 5 minutes, or about 6 minutes. In some embodiments, the first flow rate R ₁can be selected to be about 0.009 microliters/sec, about 0.01 microliters/sec, about 0.02 microliters/sec, about 0.03 microliters/sec, about 0.05 microliters/sec, about 0.1 microliters/sec, about 0.2 microliters /sec, about 0.3 microliters/sec, about 0.4 microliters/sec, or about 0.5 microliters/sec. The flow of fluid medium can be stopped for a perfusion stop period S of less than about 1 minute ₁or S ₁Can be 0 seconds. Alternatively, S ₁It may be about 30 seconds, about 1.5 minutes, about 2.0 minutes, about 2.5 minutes, or about 3 minutes. Next, a second perfusion cycle D may be performed using a different perfusion rate ₂. In some embodiments, the second perfusion rate may be higher than the first perfusion rate. In some embodiments, the second perfusion rate R ₂may be selected from about 1.0 microliters/sec, about 1.1 microliters/sec, about 1.2 microliters/sec, about 1.3 microliters/sec, about 1.4 microliters/sec, about 1.5 microliters/sec, about 1.7 microliters/sec sec, about 1.9 µl/sec, about 2.0 µl/sec, about 2.2 µl/sec, about 2.4 µl/sec, about 2.6 µl/sec, about 2.8 µl/sec, about 3.0 µl/sec , about 3.2 µl/sec, about 3.4 µl/sec, about 3.6 µl/sec, about 3.8 µl/sec, about 4.0 µl/sec, about 4.2 µl/sec, about 4.4 µl/sec, About 4.6 microliters/sec, about 4.8 microliters/sec, about 5.0 microliters/sec, about 6.0 microliters/sec, about 7.0 microliters/sec, about 8.0 microliters/sec, or about 9.0 microliters/sec. Second perfusion cycle D ₂Can be about 1 second, about 2 seconds, about 3 seconds, about 4 seconds, about 5 seconds, about 6 seconds, about 10 seconds, about 15 seconds, about 30 seconds, about 45 seconds, about 60 seconds, about 65 seconds, About 75 seconds, about 80 seconds, or about 90 seconds. The perfusion can then be stopped for a second perfusion stop period S ₂, S ₂can be about 0 seconds, about 10 seconds, about 20 seconds, about 30 seconds, about 40 seconds, about 50 seconds, about 60 seconds, about 1.5 minutes, about 1.75 minutes, about 2.0 minutes, about 2.5 minutes, about 2.75 minutes, About 3.0 minutes or about 4.0 minutes. In some embodiments, D ₁It can be about 2 hours, about 3 hours, or about 4 hours. In various embodiments, D ₁Can be about 4 hours. In various embodiments, S ₁It can be 0 seconds or less than about 1 minute. Second perfusion cycle D ₂It can be from about 1 second to about 6 seconds. In some embodiments, the second perfusion stop period S ₂It can be from about 40 seconds to about 1.5 minutes. Accordingly, there is provided a method for perfusing at least one biological cell in at least one growth chamber of a microfluidic device, comprising the steps of perfusing the at least one biological cell using a first perfusion step, the first perfusion The step comprises: subjecting a first fluid medium to a first perfusion rate R ₁flow through a flow region of the microfluidic device for a first priming time D ₁, where the flow region is fluidly connected to the growth chamber, where R ₁Selected to be a non-sweep and flow rate; stop the flow of the first fluid medium for a first perfusion stop time S ₁and repeating the first perfusion step W times, wherein W is an integer selected from 1 to 1000. The method may further comprise the step of perfusing the at least one biological cell using a second perfusion step, the second perfusion step comprising: subjecting the first fluid medium to a second perfusion rate R ₂flow for a second perfusion time D ₂, where R ₂Selected to be a non-sweep and flow rate; stop the flow of the first fluid medium for a second perfusion stop time S ₂and repeating the second perfusion step V times after the first perfusion step, where V is an integer from 1 to 1000. Second perfusion rate R ₂may be greater than the first perfusion rate R ₁. The first perfusion time D ₁May be the same as or different from the second infusion time D ₂. First perfusion stop time S ₁May be the same as or different from the second perfusion stop time S ₂. The number of repetitions W may be the same as or different from the number of repetitions V when performing the second perfusion step. R ₂Comparable R ₁At least 10 times faster. Alternatively, R ₂Comparable R ₁At least 20 times faster. R ₂Comparable R ₁At least 100 times faster. D ₁It can be from about 30 seconds to about 75 seconds. In other embodiments, D ₁It may be from about 40 minutes to about 180 minutes or from about 180 minutes to about 300 minutes. In some other embodiments, D ₁It can be from about 60 seconds to about 150 seconds. S ₁It can be from about 10 minutes to about 30 minutes. In other embodiments, S ₁It can be from about 5 minutes to about 10 minutes. In other embodiments, S ₁Can be zero. In some embodiments, D ₁can be from about 40 minutes to about 180 minutes, and S ₁Can be zero. In other embodiments, D ₁can be about 60 seconds to about 150 seconds, and S ₁It can be from about 5 minutes to about 10 minutes. In other embodiments, D ₁can be from about 180 minutes to about 300 minutes, and S ₁Can be zero. The total time for the first infusion step may be from about 1 hour to about 10 hours. In other embodiments, the total time for the first priming step may be from about 2 hours to about 4 hours. In some embodiments, W may be an integer greater than 2. In some embodiments, W can be from about 1 to about 20. In some embodiments, D ₂It can be from about 10 seconds to about 25 seconds. In other embodiments, D ₂It can be from about 10 seconds to about 90 seconds. In some embodiments, S ₂It can be from about 10 minutes to about 30 minutes. In other embodiments, S ₂Can be about 15 minutes. In some embodiments, V can be from about 10 to about 120. In some embodiments, V may be about 30 to about 50 or may be about 400 to about 500. In some embodiments, D ₂can be from about 1 second to about 6 seconds, and S ₂Can be 0 seconds. In some embodiments, D ₂can be about 10 seconds to about 90 seconds and S ₂It can be from about 40 seconds to about 1.5 minutes. In some embodiments, a total time for a repetition of one of the second priming steps may be from about 1 minute to about 15 minutes. conditioning mediumTo provide a medium (eg, a first medium or a second medium) that maintains and enhances the growth and/or viability of at least one biological cell, the first fluid medium may contain both liquid components and gaseous components (eg, gaseous components). can be dissolved in the liquid component). Additionally, the fluid medium may contain other components, such as biomolecules, vitamins, and minerals, dissolved in the liquid components. Any suitable components can be used in the fluid medium, as known to those skilled in the art. Some non-limiting examples are as discussed above, but many other medium compositions can be used without departing from the methods described herein. The medium may or may not contain serum of animal origin. In some embodiments, the fluid medium can comprise a chemically defined medium (at least prior to contacting the cells or a cell-containing fluid), and can further be a protein- or peptide-free chemically defined medium. In some embodiments, the fluid medium may comprise a reduced serum medium. The first fluidic medium can be prepared by saturating an initial fluidic medium with dissolved gas molecules prior to introduction into the microfluidic device. Additionally, saturating the initial fluidic medium with dissolved gas molecules can continue until the point in time when the first fluidic medium is introduced into the microfluidic device. Saturating the initial fluid medium can include contacting the microfluidic device with a gaseous environment capable of saturating the initial fluid medium with one of the dissolved gas molecules. Gas molecules that can saturate the initial fluid medium include, but are not limited to, oxygen, carbon dioxide, and nitrogen. The first fluid medium may further comprise moderating one of the pH of the first fluid medium. Moderating the pH of the first fluid medium may, for example, occur before and/or during the introduction of dissolved gas molecules. This relaxation can be achieved by adding a buffer. A non-limiting example of a suitable buffer is HEPES. Other buffers may be present in the medium and may or may not depend on the presence of carbon dioxide (such as a carbonation buffer system), and may be selected by the skilled artisan. Salts, proteins, carbohydrates, lipids, vitamins and other small molecules required for cell growth can also form part of the first fluid medium composition. In some embodiments, saturating the first fluid medium with gas components may be performed in a reservoir prior to introduction of the first fluid medium via the inlet. In other embodiments, saturating the first fluid medium with gas components may be performed in a gas-permeable connecting conduit between the reservoir and the inlet. In other embodiments, saturating the first fluid medium with gaseous components may be performed via a gas-permeable portion of a cover of the microfluidic device. In some embodiments, gas saturation of the fluid medium also includes maintaining humidity in the gas exchange environment such that the osmotic pressure of the fluid medium within the microfluidic device does not change during incubation. The composition of the first fluid medium may also comprise at least one secreted component from a feeding cell culture medium. Secretory feeder cell components may include growth factors, endocrine hormones, cytokines, small molecules, proteoglycans, and the like. The introduction of the at least one secreted component from the feeding cell culture medium can be performed in the same reservoir in which saturating the first fluid medium with the gaseous component is performed, or the introduction of the at least one secreted component from the feeding cell culture medium can be done before the saturation step. The first fluid medium. In some other embodiments, the composition of the first culture medium may also include one (or several) supplement(s) designed to provide a modified fluid medium for testing the response of cells to the supplement(s). Such additive(s) can, for example, enhance or reduce cell viability or growth. In some embodiments, the method can include detecting the pH of the first fluid medium as the first fluid medium is introduced through the at least one inlet. Detection of pH can be performed at a location directly proximate to one of the inlets. In some embodiments, the method can include detecting the pH of the first fluid medium while outputting the first fluid medium through an outlet. Detection of pH can be performed at a location directly proximate to one of the outlets. Either or both of the detectors used to detect pH can be an optical sensor. In some embodiments, the detector can provide an alert if the pH deviates from an acceptable range. In some other embodiments, the composition of the first fluid medium can be altered when a pH value measured by the detector deviates from an acceptable range. During the incubation step, an image of at least one growth chamber and any cells contained therein can be monitored. export at least one biological cellAfter the incubation step is complete, at least one biological cell or cell colony can be exported from the growth chamber or its isolation area. The output can include: the use of a strong dielectrophoretic (DEP) force sufficient to move one or more biological cells/cell populations. The DEP force can be actuated optically or electronically. For example, a microfluidic device can include a substrate having a DEP configuration, such as an optoelectronic tweezers (OET) configuration. In other embodiments, fluid flow and/or gravity may be used to export at least one biological cell or cell population from a growth chamber or isolation area. In other embodiments, a compressive force acting on a deformable cover region over the growth chamber or its isolation region can be used to export at least one biological cell or cell population from the growth chamber or isolation region, thereby causing the fluid to grow itself Local outflow from chamber or isolation area. After exporting the at least one biological cell or cell population from the growth chamber or isolation region, the cells can be exported from the flow region (eg, channel) out of the microfluidic device. In some embodiments, exporting the cells from the flow zone comprises using a DEP force strong enough to move one or more biological cells/cell populations. The DEP force can be generated as described above. In some other embodiments, exporting the cells from the flow region out of the microfluidic device includes using fluid flow and/or gravity to move the cells. During the exporting step, an image of at least one growth chamber and any cells contained therein can be monitored. Condition at least one surfaceIn some embodiments, the microfluidic device has at least one surface of at least one growth chamber in a conditioned state. In other embodiments, the surface of the at least one growth chamber is conditioned prior to introducing the at least one biological cell and can be performed as part of a method of culturing one or more biological cells. Conditioning the surface can include treating the surface with a conditioning agent, such as a polymer. In some embodiments, a method for treating at least one surface of at least one growth chamber of a microfluidic device (100, 300, 400, 500A to 500E, and 600) is provided, comprising the steps of: causing an excess of a conditioning agent to be included The microfluidic medium is flowed into a flow channel (FIGS. 1A-1C, 2, 3, 4A-4C); the microfluidic device is incubated for a selected period of time; and the medium in the channel is replaced. In other embodiments, a method for priming a microfluidic device comprises the steps of: flowing a priming solution containing a conditioning reagent into a flow channel; incubating the device for a selected period of time, thereby conditioning growth at least one surface of the chamber; and replacing the solution in the channel with a fluid medium. The priming solution can contain any fluid medium as described herein. The fluid medium that replaces the conditioning solution or the fluid medium with excess conditioning agent can be any medium as described herein and can additionally contain cells. In some embodiments, at least one surface can be treated with a polymerization modulating agent comprising an alkylene ether moiety. The polymerization modifier having an alkylene ether moiety can comprise any suitable alkylene ether-containing polymer including, but not limited to, any of the alkylene ether-containing polymers discussed above. In one embodiment, the surfaces of the growth chamber can be treated with amphiphilic nonionic block copolymers that contain different properties within the polymer chain (eg, Pluronic® polymers). Blocks of polyethylene oxide (PEO) and polypropylene oxide (PPO) subunits in ratio and located in different positions. Specific Pluronic® polymers used to create a conditioning surface include Pluronic® L44, L64, P85, F68 and F127 (which includes F127NF). In other embodiments, the surface can be treated with a polymerization modulating agent comprising a carboxyl moiety. Non-limiting examples of suitable carboxylic acid-containing polymerization moderators are as discussed above and any suitable carboxylic acid-containing polymerization moderator can be used to treat the surface. In other embodiments, the surface may be treated with a polymerization modulating agent comprising a carbohydrate moiety. Non-limiting examples of suitable saccharide-containing polymerization modulating agents are as discussed above and any suitable saccharide-containing polymerization modulating agent can be used to treat the surface. In other embodiments, the surface may be treated with a polymerization modulating agent comprising a sulfonic acid moiety. Non-limiting examples of suitable sulfonic acid-containing polymerization moderators are as discussed above and any suitable sulfonic acid-containing polymerization moderator can be used to treat the surface. In other embodiments, the surface may be treated with a polymerization modulating agent comprising an amino acid moiety. Non-limiting examples of suitable amino acid-containing polymerization modifiers are as discussed above and any suitable amino acid-containing polymerization modifier can be used to treat the surface. The amino acid-containing polymerization modulating reagent may comprise a protein. In some embodiments, the surface is treated with a protein, wherein the protein may comprise a component found in a mammalian serum or comprise a portion of a mammalian serum. In other embodiments, a component of mammalian serum is used to treat the surface. In some embodiments, a cell culture medium supplement such as B-27® supplement ((50x), serum free medium from ThermoFisher Scientific, Cat# 17504044) can be used to treat the surface. The mammalian serum may be fetal bovine serum (FBS). Alternatively, the mammalian serum may be calf serum (FCS). In other embodiments, the surface can be treated with a polymerization modulating agent comprising a nucleic acid moiety. Non-limiting examples of suitable nucleic acid-containing polymerization modulating agents are as discussed above and any suitable nucleic acid-containing polymerization modulating agent can be used to treat the surface. In some embodiments, a mixture of one or more polymerization modulating agents can be used to treat the surfaces of the growth chamber. In some other embodiments, the conditioning step can include treating at least one surface of the at least one growth chamber with at least one cell adhesion blocking molecule. In some embodiments, the step of treating at least one surface of at least one growth chamber with at least one cell adhesion blocking molecule may be performed prior to exporting cells from the microfluidic device. In some embodiments, the conditioning step may comprise pre-incubating the cells with at least one cell adhesion blocking molecule. In some embodiments, at least one cell adhesion blocking molecule can be used to disrupt actin fiber formation. In some embodiments, the cell adhesion blocking molecule may be cytochalasin B. In other embodiments, the at least one cell adhesion blocking molecule blocks integrin receptors. In some embodiments, the cell adhesion blocking molecule can comprise a peptide that contains an RGD motif. In some other embodiments, the at least one cell adhesion blocking molecule can attenuate the binding of cells to DNA-fouling surfaces. Cell adhesion blocking molecules that reduce cell binding to DNA-fouled surfaces can include a DNase 1 protein. In other embodiments, the at least one cell adhesion blocking molecule may comprise a small molecule of fibronectin inhibitor. In other embodiments, the at least one cell adhesion blocking molecule can be an antibody, such as an anti-B1 integrin antibody. In some embodiments, the at least one cell adhesion blocking molecule may comprise a combination of more than one type of cell adhesion blocking molecule. In other embodiments, conditioning comprises: heating the surface of the growth chamber to a temperature of about 30°C. In some embodiments, the method comprises: heating the surface to at least about 25°C, about 26°C, about 27°C, about 28°C, about 29°C, about 30°C, about 31°C, about One of 32°C, about 33°C, about 34°C, about 35°C, about 36°C, about 37°C, about 38°C, about 39°C, or about 40°C. In some embodiments, the method comprises: heating the surface to a temperature greater than about 25°C. In other embodiments, the method comprises: heating the surface to one of a range from about 30°C to about 40°C, from about 35°C to about 40°C, or from about 36°C to about 38°C temperature. In some embodiments, the method comprises: heating the surface to a temperature of at least about 30°C. In some embodiments, heating the surface includes conditioning at least one surface by treating the surface with a polymer. selection colony bodyThe methods described herein also include methods in which only one biological cell is introduced into at least one growth chamber. Provided is a method for colonizing a biological cell in a system comprising a microfluidic device having: a flow region configured to contain a flow of a first fluid medium; and at least one a growth chamber comprising an isolation region and a connecting region, the isolation region being fluidly connected to the connecting region and the connecting region including a proximal opening to the flow region, the method comprising the steps of: introducing the biological cells into the in the at least one growth chamber, wherein the at least one growth chamber is configured to have at least one surface conditioned to support cell growth, viability, portability, or any combination thereof; and at least sufficient to expand the organism The biological cells are grown for a long period of time in a colony of cells to produce a colony of biological cells. In some embodiments, the system can be any system as described herein. The microfluidic device can be any microfluidic device as described herein. In some embodiments of the method for colonizing a biological cell, the at least one conditioning surface can comprise a linking group covalently bonded to the surface, and the linking group can be linked to a structural is shaped to support a portion of cell growth, viability or transplantability of the one or more biological cells within the microfluidic device. In some embodiments, the linking group may comprise a siloxy linking group. In other embodiments, the linking group may comprise a phosphonate linking group. In some embodiments, the linking group may be indirectly linked to the moiety that is configured to support cell growth, viability, transplantability, or any combination thereof. In other embodiments, the linking group can be directly linked to the moiety that is configured to support cell growth, viability, transplantability, or any combination thereof. The linking group can be indirectly linked to the moiety configured to support cell growth, viability or mobility through attachment to a linker. In some embodiments, the linking group may be indirectly linked to the moiety configured to support cell growth, viability or mobility via attachment to a first end of a linker. In some embodiments, the linker may further comprise a linear moiety, wherein a backbone of the linear moiety comprises a backbone selected from any combination of silicon atoms, carbon atoms, nitrogen atoms, oxygen atoms, sulfur atoms, and phosphorus atoms 1 to 200 non-hydrogen atoms. In some embodiments, the backbone of the linear moiety may comprise one or more arylidene moieties. In other embodiments, the linker may comprise a triazolyl moiety. In some embodiments, the triazolyl moiety can interrupt the linear portion of the linker or can be attached to a second end of the linear portion of the linker. In various embodiments, the moiety configured to support cell growth and/or viability and/or transplantability may comprise: an alkyl or fluoroalkyl (including perfluoroalkyl) moiety; a monosaccharide or polysaccharide (which may include, but are not limited to, polydextrose); alcohols (which include, but are not limited to, propargyl alcohol); polyols, which include, but are not limited to, polyvinyl alcohol; alkylene ethers, which include ( but not limited to) polyethylene glycol; polyelectrolyte (which includes but is not limited to polyacrylic acid or polyvinylphosphonic acid); amine group (which includes its derivatives such as (but not limited to) alkylated amine groups, hydroxyalkanes alkylated amine groups, guanidine salts, and heterocyclic groups containing an unaromatic nitrogen ring atom, such as, but not limited to, morpholinyl or piperazinyl); carboxylic acids, including but not limited to propyonic acid ( which can provide a carboxylate anion surface); phosphonic acids, including but not limited to, ethynylphosphonic acid (which can provide a phosphonate anion surface); sulfonic acid anions; carboxybetaines; sulfobetaines; sulfamic acids; or amino acids. In some embodiments, the at least one conditioning surface includes an alkyl or perfluoroalkyl moiety. In other embodiments, the at least one conditioning surface includes alkylene ether moieties or polydextrose moieties. In various embodiments, the method may further comprise the step of conditioning the at least one surface of the at least one growth chamber. In some embodiments, conditioning includes treating the at least one surface with one or more agents that support cell portability within the microfluidic device. In some embodiments, the conditioning can include treating the at least one surface of the at least one growth chamber with a conditioning agent comprising a polymer. In some embodiments, the polymer may contain alkylene ether moieties. In some embodiments, the polymer may contain carboxylic acid moieties. In some embodiments, the polymer may contain carbohydrate moieties. In other embodiments, the polymer may contain sulfonic acid moieties. In other embodiments, the polymer may contain amino acid moieties. In further embodiments, the polymer may comprise nucleic acid moieties. In some embodiments, the conditioning may comprise treating the at least one surface of the at least one growth chamber with one or more components of mammalian serum. In some embodiments, the mammalian serum can be fetal bovine serum (FBS) or calf serum (FCS). In various embodiments, conditioning can include treating at least one surface of the at least one growth chamber with at least one cell adhesion blocking molecule. In some embodiments, the at least one cell adhesion blocking molecule can comprise an RGD-containing peptide. In other embodiments, the at least one cell adhesion blocking molecule can be cytochalasin B, an integrin antibody, an inhibitor of fibronectin, or a DNase 1 protein. In various embodiments, conditioning can include treating at least one surface of the at least one growth chamber with a combination of more than one type of cell adhesion blocking molecules. In various embodiments, the conditioning can include heating the at least one surface of the at least one growth chamber to a temperature of about 30°C. In various embodiments, the method may further comprise the step of introducing a first fluidic medium into a microfluidic channel of the flow region of the microfluidic device. In some embodiments, introducing the first fluid medium may be performed prior to introducing the biological cells. In some embodiments, introducing the biological cell into the at least one growth chamber can include using a dielectrophoretic (DEP) force having a force sufficient to move the biological cell. In some embodiments, the DEP force can be activated optically. In some embodiments, the DEP force may be generated by optoelectronic tweezers (OET). In some other embodiments, introducing the biological cells into the at least one growth chamber can include using fluid flow and/or gravity. In some embodiments, introducing the biological cell into the at least one growth chamber may further comprise: introducing the biological cell into an isolation region of the at least one growth chamber. In some embodiments, the isolation region of the at least one growth chamber may have sufficient capacity to support cell expansion to no more than 1 x 10 ²size of a cell. In some embodiments, the isolation region may be at least substantially filled with a second fluid medium. In some embodiments, the flow region may be fluidly connected to a proximal opening of a connection region of the at least one growth chamber, and wherein the connection region may be further fluidly connected to the isolation region of the growth chamber. In various embodiments, the method can further comprise the step of perfusing the first fluid medium during the incubation step, wherein the first fluid medium can be introduced through at least one inlet of the microfluidic device and wherein the first fluid medium can be introduced through the microfluidic device At least one outlet of the fluidic device outputs the first fluidic medium optionally including components from the second fluidic medium. In some embodiments, perfusion may be discontinuous. In some other embodiments, perfusion may be periodic. In other embodiments, perfusion may be irregular. In some embodiments, components of the second fluid medium in the isolation region may be sufficient to allow diffusion of components of the second fluid medium into the first fluid medium in the flow region and/or components of the first fluid medium to diffuse into the A rate of the second fluid medium in the isolated area performs perfusion of the first fluid medium; and the first fluid medium may not flow substantially into the isolated area. In some embodiments, perfusion of the first fluid medium may be performed about every 10 minutes to about every 30 minutes for a duration of one of about 45 seconds to about 90 seconds. In some embodiments, perfusion of the first fluid medium may be performed for a duration of one of about 2 hours to about 4 hours. In some embodiments, the time period for culturing the at least one biological cell can be from about 1 day to about 10 days. In some embodiments, a composition of the first fluid medium may include a liquid component and a gaseous component. In various embodiments, the method may further comprise the step of saturating the first fluidic medium with dissolved gas molecules prior to introducing the first fluidic medium into the microfluidic device. In various embodiments, the method may further comprise the step of contacting the microfluidic device with a gaseous environment capable of saturating the first fluidic medium or the second fluidic medium with dissolved gas molecules. In various embodiments, the method may further comprise the step of moderating the pH of one of the first fluid culture medium after introducing dissolved gas molecules. In some embodiments, saturating the first fluid medium with the gaseous components may be performed in a reservoir prior to introduction of the first fluid medium through the inlet, with the flow between the reservoir and the inlet Saturating the first fluid medium with the gaseous components is performed in a gas-permeable connector or via a gas-permeable portion of a cover of the microfluidic device. In some embodiments, a composition of the first fluid medium can comprise at least one secreted component from a feeding cell culture medium. In various embodiments, the method may further comprise the step of detecting the pH of the first fluid medium while outputting the first fluid medium through the at least one outlet. In some embodiments, the detecting step may be performed at a location directly proximate to one of the at least one outlet. In various embodiments, the method may further comprise the step of detecting the pH of the first fluid medium upon introduction of the first fluid medium through the at least one inlet. In some embodiments, the sensor may be an optical sensor. In various embodiments, the method may further comprise the step of altering a composition of one of the first fluid culture media. In various embodiments, the method may further comprise the step of monitoring an image of the at least one growth chamber and any cells contained therein. In various embodiments, the biological cell can be a mammalian cell. In some embodiments, the biological cell can be an immune cell. In some embodiments, the biological cell can be a lymphocyte or a white blood cell. In some embodiments, the biological cell can be a B cell, a T cell, a NK cell, a macrophage or a dendritic cell. In some embodiments, the biological cell can be an adherent cell. In some embodiments, the biological cell can be a fusion cell. In some embodiments, the biological cell can be a plurality of biological cells and the at least one growth chamber is a plurality of growth chambers. In various embodiments, the method may further comprise the step of moving only one of the plurality of biological cells into each of the plurality of growth chambers. In some embodiments of the method of colonizing a biological cell, the modulating surface may further comprise a lysable moiety. The method may comprise the step of lysing the lysable moiety prior to exporting one or more biological cells of the colonized population from the growth chamber or the isolation region thereof. In various embodiments, the method may further comprise the step of exporting one or more biological cells of the colony population from the growth chamber or the isolation region thereof. In some embodiments, the output may comprise: using a strong dielectrophoretic (DEP) force sufficient to move the one or more biological cells. In some embodiments, the DEP force is actuated optically. In some embodiments, the DEP force may be generated by optoelectronic tweezers (OET). In some embodiments, the output may include the use of fluid flow and/or gravity. In some embodiments, outputting may include applying a compressive force on a deformable cover region over the growth chamber or the isolation region thereof. In various embodiments, the method may further comprise the step of exporting one or more biological cells of the colonized population out of the microfluidic device from the flow region. In some embodiments, the output may comprise: using a DEP force strong enough to move the one or more biological cells. In some embodiments, the DEP force is actuated optically. In some embodiments, the DEP force may be generated by optoelectronic tweezers (OET). In some embodiments, the output may include the use of fluid flow and/or gravity. setThe present invention can provide a kit for culturing a biological cell, wherein the kit comprises: a microfluidic device having a flow region and at least one growth chamber configured to contain a flow of a first fluid medium; and a surface conditioning reagent. In this embodiment, the at least one growth chamber is not pretreated to condition at least one surface of the at least one growth chamber, but the conditioned surface is created by treatment with the surface conditioning agent prior to introduction of the cell(s). The present invention also provides other kits for culturing a biological cell, wherein the kit comprises a microfluidic device having: a flow region configured to contain a flow of a first fluid medium; and at least one a growth chamber including an isolation region and a connection region, wherein the isolation region is fluidly connected to the connection region and the connection region includes a proximal opening to the flow region; and wherein the at least one growth chamber further includes a conditioned at least one surface to support cell growth, viability, transplantability, or any combination thereof. The present invention also provides other kits for culturing a biological cell comprising a microfluidic device comprising: a flow region configured to contain a flow of a first fluid medium; and at least one a growth chamber comprising an isolation region and a connecting region, wherein the isolation region is fluidly connected to the connecting region and the connecting region has a proximal opening to the flow region; wherein the at least one growth chamber has at least one surface, It has a surface modified ligand. Alternatively, the present invention can provide a kit for culturing a biological cell, wherein the kit comprises: a microfluidic device having a flow region configured to contain a flow of a first fluid medium, and at least one growth chamber having at least one conditioning surface that can support cell growth, viability, transplantability, or any combination thereof; and a surface conditioning agent. The microfluidic device of any of the kits can be any of the

microfluidic devices

100, 200, 240, 290, 400, 500A to 500E or 600 and having any of the features described above. The microfluidic device of any of the sets can further comprise a microfluidic channel comprising at least a portion of the flow region; and the device can further comprise a growth chamber having direct access to one of the microfluidic channels connection area. The growth chamber may further include an isolation region. The isolation region can be fluidly connected to the connecting region and can be configured to contain a second fluid medium, wherein when the flow region and the at least one growth chamber are substantially filled with a first fluid medium and a second fluid medium, respectively , components of the second fluid medium diffuse into the first fluid medium and/or components of the first fluid medium diffuse into the second fluid medium; and the first medium does not flow substantially to the isolation in the area. In various embodiments of any of these sets, the growth chambers may be like the

growth chambers

124, 126, 128, 130, 244, 246, 248 or 436 general configuration, wherein the isolation region of the growth chamber can have a configuration to support no more than about 1 x 10 ³pcs, about 5×10 ²pcs, about 4×10 ²pcs, about 3×10 ²pcs, about 2×10 ²pcs, about 1×10 ²One, about 50, about 25, about 15, or about 10 volumes of cultured cells. In other embodiments, the isolation region of the growth chamber can support up to about 10, about 50, or about 1×10 ²One volume of a cell. As discussed above, any configuration of the growth chambers can be used in the growth chambers of the microfluidic devices of the sets. In various embodiments of any of the sets, the size of the growth chambers can be configured to maintain no more than 1 x 10 ²biological cells, in which the volume of these growth chambers may not exceed 1×10 ⁷cubic microns. In other embodiments, no more than 1×10 can be maintained ²biological cells, the volume of these growth chambers may not exceed 5×10 ⁶cubic microns. In other embodiments, no more than 50 biological cells can be maintained, and the volume of the growth chambers can be no more than 1 x 10 ⁶Cubic microns or not more than 5×10 ⁵cubic microns. In the sets, the microfluidic devices can have any number of growth chambers as discussed above. The microfluidic device of any of the kits can further comprise at least one inlet configured to deliver a fluidic medium (eg, a first fluidic medium or a second fluidic medium) into the flow region and configured to The fluid medium (eg, spent first fluid medium) receives at least one outlet of the fluid medium as it exits the flow region. The microfluidic device of any of the sets may further comprise a substrate having a plurality of DEP electrodes, wherein a surface of the substrate forms a surface of the growth chamber and the flow region. The plurality of DEP electrodes can be configured to generate one or more cells sufficient to move one or more biological cells (eg, a colony) into a growth chamber or the isolated region thereof or a biological cell culture medium A strong dielectrophoresis (DEP) force is removed from a growth chamber or the isolated region thereof. The DEP electrodes and thus the DEP force can be optically actuated. These optically actuated DEP electrodes can be dummy electrodes (eg, regions of an amorphous silicon substrate that have enhanced conductivity due to incident light), phototransistors, or electrodes switched on or off by a corresponding phototransistor. Alternatively, the DEP electrode and thus the DEP force may be electrically actuated. In some other embodiments, the microfluidic device may further comprise a substrate having a plurality of transistors, wherein a surface of the substrate forms a surface of the growth chamber and the flow region. The plurality of transistors are capable of producing a strong dielectric sufficient to introduce the biological cell into the growth chamber or the isolated region thereof or to remove one or more cells of a biological cell culture medium from the growth chamber or the isolated region thereof Electrophoresis (DEP) force. Each of the plurality of transistors can be actuated optically, and the DEP force can be generated by optoelectronic tweezers. The microfluidic device of any of the sets may further comprise a deformable cover region over the at least one growth chamber or isolation region thereof, whereby pressing the deformable cover region applies a pressure on one or more biological cells (eg, a colony) a force output from the growth region to the flow region. The microfluidic device of any of these sets can be configured to have a substantially gas impermeable cover. Alternatively, the entirety of a portion of the cover may be configured to be breathable. The gas permeable portion of the cover is permeable to at least one of carbon dioxide, oxygen and nitrogen. In some embodiments, the cover (or a portion thereof) is permeable to a combination of one or more of carbon dioxide, oxygen, or nitrogen. Any of the kits may further comprise a reservoir configured to contain a fluid medium. The reservoir can be fluidly connected to any of the microfluidic devices described herein. The reservoir can be configured such that the fluid medium present in the reservoir can be contacted by a gaseous environment capable of saturating the fluid medium with dissolved gas molecules. The reservoir can be further configured to contain a feeder cell population in fluid contact with the fluid medium. Any of the kits can include at least one connecting conduit configured to connect to an inlet and/or an outlet of the microfluidic device. The connecting conduit can also be configured to connect to a reservoir or a flow controller, such as a pump assembly. The connecting conduit may be breathable. The gas permeable connecting conduit is permeable to at least one of carbon dioxide, oxygen and nitrogen. In some embodiments, the gas permeable conduit is permeable to a combination of one or more of carbon dioxide, oxygen, or nitrogen. Any of the kits can further comprise a sensor configured to detect a pH of a first fluid medium. The sensor can be connected to an inlet of the microfluidic device or one of its attachment connecting conduits. Alternatively, the sensor can be integrated into the microfluidic device. The sensor can be connected proximate to the point at which the fluid medium enters the microfluidic device. The kit can include a sensor configured to detect a pH of a fluid medium at the output of the microfluidic device. The sensor can be connected to an outlet of the microfluidic device or one of its attachment connecting conduits. Alternatively, the sensor can be integrated into the microfluidic device. The sensor can be connected proximate to the point at which the fluid medium exits the microfluidic device. Whether or not the sensor is attached to the inlet and/or the outlet of the microfluidic device, it can be an optical sensor. An optical sensor may include an LED and an integrated colorimetric sensor, which may optionally be a color-sensitive phototransistor. The kit may further include drive electronics to control the pH sensor and receive output from the pH sensor. The kit may further comprise a pH detection agent. The pH detection agent may be a pH sensitive dye detectable under visible light. Any of the kits may also include a culture medium having components capable of enhancing the viability of biological cells on the microfluidic device. These components can be any suitable medium components known in the art, including any of the components discussed above for fluid medium components. Any of the kits can further comprise at least one reagent for detecting a state of a biological cell or a population of cells. Reagents configured to detect the state of a cell are well known in the art and can be used, for example, to detect whether a cell is alive or dead, secreting a substance of interest such as an antibody, cytokine or growth factor), or whether it has the cell surface marker of interest. Such reagents can be used in, but are not limited to, the kits and methods described herein. For any of the kits provided herein, the components of the kits can be located in separate containers. For any of the components of the kit provided in solution, the components may be present at a concentration of about 1, 5, 10, 100 or 1000 times the concentration used in the methods of the present invention . For a kit in which at least one growth chamber of a microfluidic device is not pretreated to condition at least one surface of at least one growth chamber, a conditioned surface is produced by treating with a surface conditioning agent; or for a kit comprising a microfluidic device having a : a flow region configured to contain a flow of a first fluid medium; and at least one growth chamber having at least one conditioning surface that can support cell growth, viability, portability, or any combination thereof ) and a set of surface conditioning reagents that can be used to precondition the surface of the growth chamber. The surface conditioning agent may comprise a polymer, which may be any one or more of the polymers described above for use as a surface conditioning agent. In some embodiments, the surface modifying agent may comprise a polymer having an alkylene ether moiety, a carboxylic acid moiety, a sulfonic acid moiety, an amino acid moiety, a nucleic acid moiety, a carbohydrate moiety, or any combination thereof. The surface conditioning agent may comprise a PEO-PPO block copolymer, such as a Pluronic® polymer (eg L44, L64, P85 or F127). In some embodiments, the surface modulating agent may comprise one or more components of mammalian serum. The mammalian serum can be fetal bovine serum (FBS) or calf serum (FCS). Alternatively, the surface conditioning reagent used to condition the surface of the growth chamber may be included in the kit, separate from the microfluidic device. In other embodiments of the kit, a preconditioning microfluidic device and a surface conditioning reagent different from the surface conditioning reagent used to condition the surface of the growth chamber are included. The different surface-modulating reagents can be any of the surface-modifying reagents discussed above. In some embodiments, more than one surface modulating agent is included in the kit. In various embodiments of the kits with a microfluidic device in which the at least one growth chamber of the microfluidic device is not pretreated to condition the at least one surface, the kits may also include a device suitable for culturing the A culture medium of one or more biological cells. In some embodiments, the kit may also include a medium supplement including an agent capable of enhancing the conditioning of a surface of the growth chamber. The medium supplement may comprise a modulating agent as discussed above or another chemical species that enhances the ability of the at least one surface of the at least one growth chamber to support cell growth, viability, transplantability, or any combination thereof. This may include growth factors, endocrine hormones, antioxidants or vitamins and the like. The kit may also include a flow controller configured to perfuse at least the first fluid medium, which may be a separate component of the microfluidic device or may be incorporated as part of the microfluidic device. The controller can be configured to discontinuously perfuse the fluid medium. Thus, the controller can be configured to perfuse the fluid medium in a periodic pattern or in an irregular pattern. In another aspect, a kit for culturing a biological cell is provided, comprising a microfluidic device having: a flow region configured to contain a flow of a first fluid medium; and at least a growth chamber including an isolation region and a connection region, wherein the isolation region is fluidly connected to the connection region and the connection region includes a proximal opening to the flow region; and wherein the at least one growth chamber further includes a At least one surface modulated to support cell growth, viability, transplantability, or any combination thereof. The microfluidic device can be any microfluidic device as described herein, and can have any of the growth chambers as described herein. The microfluidic device can have a substrate having a DEP configuration of any of the kinds described herein. The DEP configuration can be actuated optically. The substrate of the microfluidic device may have a surface comprising a substrate composition of Formula 1 or Formula 2 (as described herein) and having all of the features as described above.

Equation 1 Equation 2 The at least one conditioning surface of the microfluidic device of the kit may comprise a carbohydrate moiety, an alkylene ether moiety, an amino acid moiety, an alkyl moiety, a fluoroalkyl moiety (which may include a perfluoroalkyl moiety), an anion moiety, cationic moiety and/or zwitterionic moiety. In some embodiments, the conditioning surface of the microfluidic device may comprise a carbohydrate moiety, an alkylene ether moiety, an alkyl moiety, a fluoroalkyl moiety, or an amino acid moiety. The alkyl or perfluoroalkyl moieties can have a backbone length of greater than 10 carbons. In some embodiments, the modulating surface to support cell growth, viability, transplantability, or any combination thereof, may comprise: an alkyl or fluoroalkyl (including perfluoroalkyl) moiety; a monosaccharide or Polysaccharides (which may include, but are not limited to, polydextrose); alcohols (which include, but are not limited to, propargyl alcohol); polyols, which include, but are not limited to, polyvinyl alcohol; alkylene ethers, which include (but not limited to) polyethylene glycol; polyelectrolytes (including but not limited to polyacrylic acid or polyvinylphosphonic acid); amine groups (including but not limited to derivatives thereof, such as (but not limited to) alkylated amine groups, hydroxyl groups Alkylated amine groups, guanidine salts, and heterocyclic groups containing an unaromatic nitrogen ring atom such as (but not limited to) morpholinyl or piperazinyl); carboxylic acids, including but not limited to propyonic acid (which can provide a carboxylate anion surface); phosphonic acids, including but not limited to, ethynylphosphonic acid (which can provide a phosphonate anion surface); sulfonic acid anions; carboxybetaines; sulfobetaines; sulfamic acids ; or amino acids. In some embodiments of the kit, the conditioning surface can include a linking group covalently bonded to a surface of the microfluidic device, and the linking group can be linked to a configuration to support the The portion of the cell growth, viability, transplantability, or any combination thereof, of the one or more biological cells within the microfluidic device. The linking group may be a siloxy linking group. Alternatively, the linking group may be a phosphonate linking group. In some embodiments of the kit, the linking group of the modulating surface can be directly linked to the moiety that is configured to support cell growth, viability, transplantability, or any combination thereof. In other embodiments, the linking group may be indirectly linked via a linker to the moiety configured to support cell growth, viability, transplantability, or any combination thereof, or the like. The linking group can be indirectly linked to the moiety configured to support cell growth, viability, transplantability, or any combination thereof, via attachment to a first end of a linker. The linker may further comprise a linear moiety, wherein a main chain of the linear moiety comprises 1 to 200 non-ferrous atoms selected from any combination of silicon atoms, carbon atoms, nitrogen atoms, oxygen atoms, sulfur atoms and phosphorus atoms A hydrogen atom. In some embodiments of the kit, the linker of the conditioning surface may further comprise a triazolyl moiety. The cleavable moiety is configured to allow destruction of the regulatory surface, thereby promoting the biomolecule's portability. The kit may further comprise an agent configured to cleave the cleavable portion of the modulating surface. In various embodiments of the kit, the kit can further comprise a surface conditioning agent. In some embodiments, the surface modifying agent may comprise a polymer comprising at least one of the following: an alkylene ether moiety, a carboxylic acid moiety, a sulfonic acid moiety, a phosphonic acid moiety, an amino acid moiety, a nucleic acid moiety, or sugar part. In some other embodiments, the surface modifying agent comprises a polymer comprising at least one of the following: an alkyl ether moiety, an amino acid moiety, or a carbohydrate moiety. In some other embodiments, the conditioning surface may comprise a cleavable moiety. In other embodiments of the kit, the surface modulating agent comprises at least one cell adhesion blocking molecule. In some embodiments, the at least one cell adhesion blocking molecule disrupts actin fibril formation, blocks integrin receptors, or reduces cell binding to DNA-fouling surfaces. In some embodiments, the at least one cell adhesion blocking molecule can be cytochalasin B, an RGD-containing peptide, a DNase 1 protein, a fibronectin inhibitor, or an integrin antibody. In some embodiments, the at least one cell adhesion blocking molecule may comprise a combination of more than one type of cell adhesion blocking molecule. In various embodiments of the kit, the surface modulating agent may comprise one or more components of mammalian serum. The mammalian serum can be fetal bovine serum (FBS) or calf serum (FCS). In various embodiments of the kit, the kit can further comprise a medium suitable for culturing the one or more biological cells. In some embodiments, the kit can include a medium supplement comprising an agent configured to enhance conditioning of the at least one surface of the growth chamber. The medium supplement may comprise a modulating agent as discussed above or another chemical species that enhances the ability of the at least one surface of the at least one growth chamber to support cell growth, viability, transplantability, or any combination thereof. This may include growth factors, endocrine hormones, antioxidants or vitamins and the like. In various embodiments of the kit, the kit can include at least one reagent for detecting a state of the one or more biological cells. In yet another aspect, a kit for culturing a biological cell is provided, comprising a microfluidic device for culturing one or more biological cells, the microfluidic device comprising: a flow region configured with a flow containing a first fluid medium; and at least one growth chamber comprising an isolation region and a connecting region, wherein the isolation region is fluidly connected to the connecting region and the connecting region has a proximity to a region of the flow region The end is open; and the at least one growth chamber has at least one surface with a surface modification ligand. The microfluidic device can be any microfluidic device as described herein. The surface may comprise a substrate having a dielectrophoretic (DEP) configuration. The DEP configuration can be any of the DEP configurations described herein. The DEP configuration can be actuated optically. The substrate is any substrate having a surface-modified ligand as described herein, and may have a structure of formula 3, and may include all of the features as described above:

Formula 3 In various embodiments of the kit having a microfluidic device having at least one surface comprising a surface-modifying ligand, the surface-modifying ligand can be covalently bonded to the surface of the substrate the oxide part. The surface-modifying ligand may comprise a reactive moiety. The reactive moiety of the surface modification ligand can be an azide, amine, bromo, thiol, active ester, succinimidyl or alkynyl moiety. The surface-modifying ligands can be covalently linked to the oxide moieties via a linking group. In some embodiments, the linking group can be a siloxy moiety. In other embodiments, the linking group may be a phosphate moiety. The linking group can be indirectly attached to the reactive moiety of the surface-modifying ligand via a linker. The linker may comprise a linear moiety, wherein a backbone of the linear moiety comprises 1 to 100 non-hydrogens selected from any combination of silicon atoms, carbon atoms, nitrogen atoms, oxygen atoms, sulfur atoms and phosphorus atoms atom. In some embodiments, the surface-modifying ligand may comprise one or more cleavable moieties. The one or more lysable moieties can be configured to allow destruction of a conditioned surface of a microfluidic device after forming the conditioned surface, thereby promoting the portability of the one or more biological cells after culture. In some embodiments of the kit having a microfluidic device having at least one surface comprising a surface-modifying ligand, the kit can further comprise a modulating modifier comprising a modulating modifier configured to A first moiety that supports cell growth, viability, transplantability, or any combination thereof, and a second moiety configured to react with the reactive moiety of the surface-modifying ligand, the modulating modifier may have Any of the structures of Formula 5 and having the features as described herein:

Formula 5 The second moiety can be configured to convert the surface-modifying ligand into a configuration to support the growth chamber after reacting with the reactive moiety of the surface-modifying ligand of the microfluidic device of the set One of the cell growth, viability, transplantability, or any combination thereof of one or more biological cells modulates the surface. The first moiety may comprise an alkylene oxide moiety, a carbohydrate moiety, an alkyl moiety, a perfluoroalkyl moiety, an amino acid moiety, an anionic moiety, a cationic moiety, or a zwitterionic moiety. In some embodiments, the first moiety may comprise: an alkyl or fluoroalkyl (which includes perfluoroalkyl) moieties; a monosaccharide or polysaccharide (which may include, but is not limited to, polydextrose); an alcohol (including ( but not limited to) propargyl alcohol); polyols including but not limited to polyvinyl alcohol; alkylene ethers including but not limited to polyethylene glycol; polyelectrolytes including but not limited to polyacrylic acid or polyvinylphosphonic acid); amine groups (which include derivatives thereof such as, but not limited to, alkylated amine groups, hydroxyalkylated amine moieties, guanidine salts, and heterocyclic compounds containing an unaromatic nitrogen ring atom) Cyclic groups such as, but not limited to, morpholinyl or piperazinyl); carboxylic acids, including but not limited to propynoic acid (which can provide a carboxylate anion surface); phosphonic acids, including but not limited to ) ethynylphosphonic acid (which can provide a phosphonic acid anion surface); sulfonic acid anion; carboxybetaine; sulfobetaine; amine sulfonic acid; or amino acid. The second moiety can be an amine, carboxylic acid, alkynyl, azide, aldehyde, bromo or thiol moiety. In some embodiments, the first moiety or a linkage L' of the modulating modifier (as described above for Formula 5) can comprise a cleavable moiety. The cleavable moiety can be configured to allow destruction of the regulatory surface, thereby promoting the transplantability of the biological cells. In some embodiments, the kit can further comprise an agent configured to cleave the cleavable portion of the modulating surface. In some embodiments of the kit having a microfluidic device having at least one surface comprising a surface-modifying ligand, the kit can further comprise a surface-modifying agent. In some embodiments of the kit having a microfluidic device having at least one surface comprising a surface-modifying ligand, the surface-modifying agent can comprise a polymer comprising at least one of the following: Alkylene ether moiety, carboxylic acid moiety, sulfonic acid moiety, phosphonic acid moiety, amino acid moiety, nucleic acid moiety or carbohydrate moiety. In some other embodiments, the surface modifying agent comprises a polymer comprising at least one of an alkylene ether moiety, an amino acid moiety, or a carbohydrate moiety. In some other embodiments, the conditioning surface may comprise a cleavable moiety. In some embodiments of the kit having a microfluidic device having at least one surface comprising a surface-modifying ligand, the surface modulating agent includes at least one cell adhesion blocking molecule. In some embodiments, the at least one cell adhesion blocking molecule disrupts actin fibril formation, blocks integrin receptors, or reduces cell binding to DNA-fouling surfaces. In some embodiments, the at least one cell adhesion blocking molecule can be cytochalasin B, an RGD-containing peptide, a DNase 1 protein, a fibronectin inhibitor, or an integrin antibody. In some embodiments, the at least one cell adhesion blocking molecule may comprise a combination of more than one type of cell adhesion blocking molecule. In some embodiments of the kit having a microfluidic device having at least one surface comprising a surface-modifying ligand, the surface modulating reagent may comprise one or more components of mammalian serum. The mammalian serum can be fetal bovine serum (FBS) or calf serum (FCS). In some embodiments of the kit having a microfluidic device having at least one surface comprising a surface-modifying ligand, the kit can further comprise a material suitable for culturing the one or more biological cells a medium. In some embodiments, the kit can further comprise a medium supplement comprising an agent configured to enhance conditioning of the at least one surface of the growth chamber. The medium supplement may comprise a linker as discussed above or another chemical species that enhances the ability of the at least one surface of the at least one growth chamber to support cell growth, viability, transplantability, or any combination thereof. This may include growth factors, endocrine hormones, antioxidants or vitamins and the like. In some embodiments of the kit having a microfluidic device having at least one surface comprising a surface-modified ligand, the kit can further comprise a device for detecting the one or more biological cells At least one reagent in a state. Example example 1. one K562 red and white blood sick Cell culture and growth Material:K562 cells (a human immortalized myeloid leukemia cell line) were obtained from the American Type Culture Collection (ATCC) (catalog ATCC® CCl-243 ^TM) and provided as a suspension cell line. By seeding 1×10 ³viable cells/mL and maintained in culture at 37°C using a 5% carbon dioxide gas environment. Divide cells into 1×10 ⁶cells/mL or divide cells every 2 to 3 days. Freeze cells in 5% dimethylsulfoxide (DMSO)/95% complete growth medium. Medium:Iscove's Modified Dulbecco's Medium (ATCC® Catalog No. 30-2005) + 10% Fetal Bovine Serum (Hyclone, Cat# SH30071.2) was combined to create a complete growth medium. When perfused during the incubation cycle, the complete growth medium was continuously conditioned using 5% carbon dioxide in air prior to its introduction into the microfluidic device. Prefill solution:Complete Growth Medium with 0.1% Pluronic® F127 (Life Technologies® Cat# P6866) Systems and Microfluidic Devices:Manufactured by Berkeley Lights. The system includes at least one flow controller, temperature controller, fluid medium conditioning and pumping components, light source for light-activated DEP configuration, microfluidic device, mounting station, and a camera. The growth chamber of the microfluidic device used in this experiment has a size of about 1.4×10 ⁵A volume of cubic micrometers. The cross-sectional area of the flow channel is about 4×10 ³square microns. The microfluidic device has 8 channels. Culture preparation: Load the microfluidic device onto the system and flush the microfluidic device with 100% carbon dioxide with 15 psi for 5 minutes. Following the carbon dioxide flush, the priming solution was instantly perfused through the microfluidic device at 5 μΐ/sec for 8 minutes. Next, complete growth medium was flowed through the microfluidic device at 5 microliters/sec for 5 minutes. Culture conditions: makeThe temperature of the microfluidic device was maintained at 37°C. The medium was perfused at a constant rate of 0.001 [mu]l/sec throughout the period of the culture experiment. Gravity was used to load a single K562 cell into one of the growth chambers of the microfluidic device. A photograph of one of the growth chambers at t=0 hours after loading cells is shown (see Figure 10A). Arrow 1002 points to the location of a single cell in the growth chamber. After the 16 hour culture was complete, the cells were expanded to a population of 2 cells, as shown in one of the photographs taken at this time point (see Figure 10B). Arrows 1004 point to the location of the two cells in the growth chamber. After the 34 hour culture was complete, the cell population was increased to a total of four cells, as shown in the photograph of Figure 1OC.

Arrows

1006 and 1008 point to each of the two groups of two cells positioned within the growth chamber. After the 54 hour culture was complete, the K562 cell population was increased to a total of eight cells, as shown in the photograph of Figure 10D.

Arrows

1010 and 1012 point to cells positioned at either side of the cell population within the growth chamber. After the 70 hour culture was complete, the K562 cell population was increased to a total of 16 cells, as shown in the photograph of Figure 1OE.

Arrows

1014, 1016 and 1018 point to the cells of the group. A colonized expanded population of K562 was provided in the growth chamber of the microfluidic device. Example 2.one OKT3 Culture and growth of fusion tumor cells Material:OKT3 cells, a murine myeloid fusionoma cell line, were obtained from ATCC (ATCC® Cat. # CRL-8001™). Cells were provided as a suspension cell line. By seeding about 1 x 10 ⁵viable cells/mL to about 2 x 10 ⁵Viable cells/mL and maintained in culture at 37°C using 5% carbon dioxide in air as a gaseous environment. Cells were allowed to divide every 2 to 3 days. Count the number and viability of OKT3 cells and adjust the cell density to 5×10 ⁵/ml to load into microfluidic devices. Medium: 500 mL Iscove's Modified Dulbecco's Medium (ATCC® Catalog No. 30-2005), 200 mL Fetal Bovine Serum (ATCC® Cat. #30-2020), and 1 mL Penicillin-Streptomycin (Life Technologies® Cat. # 15140-122) were combined to produce media. Complete medium was filtered through a 0.22 μm filter and stored at 4°C in the dark until use. When perfused during the incubation cycle, the medium was continuously conditioned using 5% carbon dioxide in air prior to introduction into the microfluidic device. Precharge Solution:Medium containing 0.1% Pluronic® F127 (Life Technologies® Cat# P6866). Systems and Microfluidic Devices:Manufactured by Berkeley Lights. The system includes at least one flow controller, temperature controller, fluid medium conditioning and pumping components, light source and projector for light-activated DEP configuration, microfluidic device, mounting table, and a camera. The growth chamber of the microfluidic device used in this experiment has a size of about 1.5×10 ⁶A volume of cubic micrometers. The cross-sectional area of a flow channel is 8×10 ³square microns, and a total of six channels exist on the microfluidic device. Cultivation preparation:Microfluidic devices were loaded onto the system and 100% carbon dioxide with 15 psi was used to flush the microfluidic devices for 5 minutes. Following the carbon dioxide flush, the priming solution was instantly perfused through the microfluidic device at 8 μl/sec until a total volume of 2.5 mL was perfused through the microfluidic device. Next, the medium was flowed through the microfluidic device at 8 μΐ/sec until a total of 1 mL of medium was perfused through the microfluidic device. Preparation of the microfluidic device prior to introduction of cells is shown in the photograph of Figure 11A. A column of four growth chambers extends along the bottom of the photo. Culture conditions:The temperature of the microfluidic device was maintained at 37°C. The medium was perfused throughout the cycle of the culture experiment using a variable perfusion method comprising an initial 4-hour perfusion cycle at 0.01 μl/sec followed by a short high-speed at 8 μl/sec Perfusion (about 3 seconds) followed by a short perfusion off period of about less than 1 minute. This cycle, including alternating perfusion rates and a stop, continued throughout the culture experiment. A single OKT3 cell was introduced into the growth chamber by gravity. A photograph of a growth chamber with one cell at time t=0 is shown in FIG. 11B, where arrow 1102 points to the second chamber from the left, and in particular to a single cell within the chamber, where the area of resident cells is further circled. 12A-12C show photographs of microfluidic devices at subsequent time points in culture experiments and demonstrate cell expansion to form a colony. The photograph of Figure 12A was taken at the end of one day of culture and arrow 1202 points to a population of about 4 cells in the second chamber from the left (the point of introduction of single OKT3 cells). Figure 12B is a photograph taken after the 2 day culture is complete and arrow 1204 points to one of the further proliferating cell populations in the second chamber on the left. Figure 12C is a photograph taken after the 3 day culture was completed, and arrow 1206 shows the numerous expanded OKT3 cells resulting from culturing a single OKT3 cell. Figures 13A-13C show photographs of the microfluidic device after the three-day culture is complete (ie, after the time point of Figure 12C) and demonstrate the use of a dielectrophoretic force generated by optoelectronic tweezers to output a selection of expanded OKT3 cells . The light pattern that induces the dielectrophoretic force (ie, a light trap pointed to by arrow 1302) is shown in Figure 13A as a white box surrounding the cell. Cells are moved from the bottom of the growth chamber towards the flow channel by optically actuated dielectrophoretic forces. The photograph of Figure 13B shows that the expanded OKT3 cells moved further into the flow area. The cells are still trapped in the optical trap and are forced to move with the optical trap (arrow 1304). The photograph of Figure 13C shows the release of the expanded cells (arrow 1306) after they have completely moved into the flow area. These cells are exported from the microfluidic device for further study or expansion by using optically actuated DEP force, gravity or fluid flow. This experiment demonstrates the selectivity, precision, and flexibility provided by the use of the devices and methods described herein. example 3. Using a serum-free medium to remove adherent cells to condition the surface of a microfluidic device Systems and Microfluidic Devices:As in Example 1, using a ⁵A growth chamber with a volume of one cubic micrometer. Pre-charge plan:Inject 250 microliters of 100% carbon dioxide at a rate of 12 microliters/sec. Thereafter, 250 microliters of PBS containing 0.1% Pluronic® F27 (Life Technologies® Cat# P6866) was flowed in at 12 microliters/sec. The final step of priming consisted of influxing 250 microliters of PBS at 12 microliters/sec. Next, the medium is introduced. Perfusion scheme:The perfusion method is either of the following two methods: 1. Perfuse at 0.01 μl/sec for 2 hours; perfuse at 2 μl/sec for 64 seconds; and repeat. 2. Perfuse at 0.02 μl/sec for 100 seconds; stop flow for 500 seconds; perfuse at 2 μl/sec for 64 seconds; and repeat. culture medium :Serum-free medium (ThermoFisher Scientific, Cat. No. 12045-096). Systems and Microfluidic Devices :The ability to remove adherent cells from the flow channel of a microfluidic device after incubation was demonstrated at 36°C with a conditioned medium supplement (B-27® supplement) (2% v/v) without a Adherent cells, which can be, for example, JIMT1 cells purchased from AddexBio, Cat. No. C000605, are pre-incubated in serum medium for 30 minutes. After pre-incubation, the adherent cells are introduced into the flow channel, the flow is stopped, and the adherent cells are incubated for a period of 2 hours to about 24 hours. After the assay was complete, a flow of serum-free medium was introduced at a rate of 5 microliters/second. A flow rate of about 750 microliters (which represents about 150 times the volume of the microfluidic device) was passed through the microfluidic device, and all attached JIMT1 cells were output from the flow channel and exited the microfluidic device. This experiment shows that serum-free medium, which may contain supplemental components such as commercially available B27, prevents sticking during the process of an assay incorporating adherent reporter cells, and allows the export of adherent cells from the microfluidic device. Example 4. Using a Conditioning Mixture to Remove Adherent Cells to Condition the Surface of a Microfluidic Device Attached cells:As in Example 3 above. Medium:One serum-free medium ( ThermoFisher Scientific, Cat. No. 12045-076). Adjust the mix thing :Cytochalasin B (Sigma Aldrich, Catalog No. C2743-200UL), DNaseI (New England Biosciences Cat No. M0303S) and RGD tripeptide (Santa Cruz Biotechnology Cat No. sc-201176). Adherent cell preparation:The medium was modified using the conditioning mixture to have one of the following final concentrations: 4 micromolar cytochalasin B, 0.1 units/micromolar DNaseI, and 1 mM RGD tripeptide. Adherent cells were incubated at 36°C for 30 minutes prior to infusion into the microfluidic device. Systems and Microfluidic Devices.As above, using about 7 x 10 ⁵A growth chamber with a volume of one cubic micrometer. The ability to remove adherent cells (eg, JIMT1 cells) from the flow channel of a microfluidic device after incubation is demonstrated by pre-incubating a population of adherent cells pre-incubated with a conditioning mixture. Notably, the use of conditioning mixtures allows serum-containing media, such as those used in this example, to be used within a microfluidic environment, while still allowing for the removal of adherent cells. The pre-incubated adherent cells are introduced into the flow channel of the microfluidic device, and the adherent cells are incubated for a period of 2 hours to about 24 hours. After the assay is complete, a flow of medium is introduced at a rate of 5 microliters/second. A flow rate of about 750 microliters (which represents about 150 times the volume of the microfluidic device) was passed through the microfluidic device, and all attached cells were output from the flow channel and exited the microfluidic device. This experiment shows that conditioning the mixture prevents adhesion and allows export of adherent cells. example 5. Fabrication of microfluidic devices with regulated surfaces For all preparations:Microfluidic device: As in Example 1 above, manufactured by Berkeley Lights and used as received. In either case, silicon substrates with patterned polysiloxane (PPS) and ITO/glass substrates were subjected to a Nordson Asymtek plasma prior to synthesis of conditioning surfaces cleanerOxygen plasma for medium cleaning (100 W power, 50 s). A. perfluoroalkylsiloxane base conditioning surface Material:Heptadecafluoro-1,1,2,2-tetrahydrododecyltrimethoxysilane was obtained from Gelest (Cat. No. SIH5841.5) and used directly. Use MgSO directly ₄·7H ₂O (Acros). Preparation method:Chemical modification by exposing the assembled microfluidic device to heptadecafluoro-1,1,2,2-tetrahydrododecyltrimethoxysilane and water vapor at elevated temperature under reduced pressure Assembled Microfluidic Devices. Add 300 μl heptadecafluoro-1,1,2,2-tetrahydrododecyltrimethoxysilane and 0.5 g MgSO ₄·7H ₂O (water source) to separate the aluminum boat in the bottom of a clean dry 6" glass vacuum desiccator. The microfluidic device is supported on a porous plate above the silane reagent and hydrated salt (water source). The desiccator was pumped to 750 mTorr at room temperature and the desiccator was sealed. Next, the desiccant was placed in a 110°C oven for 24 hours. Next, the microfluidic device with the perfluoroalkyl conditioned surface was removed from the desiccator and used. In some experiments, the microfluidic device was chemically modified prior to mounting the microfluidic device to a printed circuit board. b. polydextrose conditioning surface Material:11-Azidoundecyltrimethoxysilane was synthesized from 11-bromoundecyltrimethoxysilane (Gelest) by replacing the bromide moiety with sodium azide. In a typical reaction, 4.00 grams of 11-bromoundecyltrimethoxysilane (Gelest) was added to 60 microliters of anhydrous dimethylformamide ( DMF) (Acros) solution. The solution was stirred at room temperature under nitrogen for 24 hours. Next, the solution was filtered and the filtrate was extracted using dry pentane (Acros). The crude 11-azidoundecyltrimethoxysilane product was concentrated by rotary evaporation and purified by two consecutive vacuum distillations. Dibenzocyclooctyne (DBCO) modified polydextrose (MW: about 3000 Da) was purchased from Nanocs and used directly. Preparation method : A surface modification ligand is introduced.The surface of the assembled microfluidic device was chemically modified by exposing the surface of the assembled microfluidic device to 11-azidoundecyltrimethoxysilane and water vapor at elevated temperature under reduced pressure. Add 300 μl 11-azidoundecyltrimethoxysilane and 0.5 g MgSO ₄·7H ₂O (water source) to separate the aluminum boat in the bottom of a clean and dry 6'' glass vacuum dryer. The microfluidic device was supported on a porous plate above the silane and hydrated salt (water source). The desiccator was pumped to 750 mTorr at room temperature and the desiccator was sealed. Next, place the dryer in a 110°C oven for 24 hours. Next, the microfluidic wafers with the surface-modified ligands (11-azidoundecylsiloxy moieties) were removed from the desiccator. In some experiments, the microfluidic device was chemically modified prior to mounting the microfluidic device to a printed circuit board. Introduction of Polydextrose Conditioning Surface:Azide-terminated microfluidic device surface by flowing at least 250 microliters of an aqueous solution containing 166 micromoles of DBCO polydextrose through the microfluidic device with surface-modified azide ligands after vapor deposition Reacts with DBCO polydextrose. The reaction was allowed to proceed at room temperature for at least 1 hour. Next, rinse the wafer by flowing at least 250 microliters of DI water through the wafer. c. polyethylene glycol (PEG) conditioning surface Material :11-Azidoundecyltrimethoxysilane was synthesized as above. Alkynyl-modified PEG (MW: about 5000 Da) was purchased from JenKem and used directly. Sodium ascorbate and copper sulfate pentahydrate were purchased from Sigma-Aldrich and used as received (THPTA (tris(3-hydroxypropyltriazolylmethyl)amine) copper catalyzed click reagent (Glen Research)). method of preparation : A surface modification ligand is introduced.Microfluidic wafers with 11-azidoundecylsiloxy surface-modified ligands were prepared as above. PEG The introduction of conditioning surfaces:By flowing at least 250 microliters of an aqueous solution containing 333 micromoles of alkynyl-modified PEG, 500 micromoles of copper sulfate, 500 micromoles of THPTA ligand, and 5 mM of sodium ascorbate through a solution with 11-azide The undecylsiloxy surface modifies the ligand-based microfluidic device such that the azide-terminated surface of the microfluidic device reacts with the alkynyl-modified PEG. The reaction was allowed to proceed at room temperature for at least 1 hour. Next, the microfluidic device with a PEG conditioned surface was rinsed by flowing at least 250 microliters of deionized water through the device. D. Alkyl Modified Surfaces with Phosphonate Linking Groups to the Surface Material :Stearyl phosphonate was purchased from Sigma Aldrich and used as received. Acetone and ethanol were purchased from Sigma Aldrich. Preparation method:The surface of the microfluidic device was exposed to a 10 mM solution of stearyl phosphonate in absolute ethanol for 48 hours at 35°C. The resulting microfluidic devices with alkyl modulating surfaces attached via phosphonate linking groups were thoroughly rinsed with ethanol and DI water after deposition. Example 6. a modulated microfluidic surface T Culture and export of lymphocytes Material:CD3+ cells from AllCells were mixed with anti-CD3/anti-CD28 magnetic beads (Dynabeads®, Thermofisher Scientific, Cat. No. 11453D) at a rate of one bead/1 cell. at -5% CO at 37°C ₂The mixture was incubated for 5 hours in the incubator in the same medium as the culture experiment itself. After incubation, the T cell/bead mixture was resuspended for use. Medium:RPMI-1640 (GIBCO®, ThermoFisher Scientific, Cat. No. 11875-127), 10% FBS, 2% human AB serum (50 U/ml IL2; R&D Systems) Precharge program:As in Example 3 above. Perfusion scheme:As in Example 3 above. Systems and Microfluidic Devices:As in Example 3 above. The growth chamber has about 7 x 10 ⁵A volume of cubic micrometers. Conditioning Surface:The microfluidic device has a covalently linked polydextrose modulating surface prepared as described above. The (T cell + beads) suspension was introduced into the microfluidic device by flowing the resuspension through a fluidic inlet and into the microfluidic channel. The flow was stopped and the T cells/beads were randomly loaded into the growth chamber by tilting the wafer and allowing the mobile T cells/beads to be pulled into the growth chamber. After T cells/beads were loaded into the growth chamber, media was perfused through the microfluidic channels of the nanofluidic wafer for a time period of 4 days. Figure 14A shows that polydextrose modulates the growth of T cells on the surface of the growth chamber of the microfluidic device. The growth of T cells on a polydextrose regulated surface was improved relative to a non-regulated surface of a similar microfluidic device (data not shown). Next, the T cells are removed from the growth chamber by gravity (eg, tilting the microfluidic device). Figure 14B shows the extent of removal from the growth chamber at the end of a 20 minute period (which demonstrates the superior ability to export expanded T cells into the flow channel) compared to from a non-regulated surface of a similar microfluidic device Improved by the extent to which T cells are removed. Next, the T cells were removed from the microfluidic device (not shown). The examples shown herein are illustrative and in no way limit the scope of the methods and apparatuses described in the [Embodiments].

100:微流體器件 102:封閉體/蓋 104:支撐結構/基底 106:流動路徑 107:端口 108:微流體線路結構 109:內表面 110:蓋 114:框架 116:微流體線路材料 120:微流體線路 122:微流體通道 124:微流體生長室/入口 124':出口 126:微流體生長室 128:微流體生長室 130:微流體生長室 132:微小物體捕集器 134:側通路/流動通道/流動區域 150:系統 152:控制及監測設備 154:主控制器 156:控制模組 158:記憶體 160:培養基模組 162:動力模組 164:成像模組 166:傾斜模組 168:其他模組 170:顯示器件 172:輸入/輸出器件/控制模組 178:培養基源 180:流體培養基/控制/監測設備 190:傾斜器件 192:電源 194:成像器件 200:微流體器件 202:區域/室/流體培養基 202':廢流體培養基 202a':染色流體培養基 202a'':染色出流流體培養基 204:底部電極/流體培養基 206:電極啟動基板 208:內表面 210:頂部電極 212:電源/流體流 214:未經照射之介電泳(DEP)電極區域 214a:經照射之介電泳(DEP)電極區域 220:光源 222:光圖案 224:正方形圖案 226:控制器 240:微流體器件 242:端口 244:生長室 246:生長室 248:生長室 250:隔離結構 252:近端開口 254:連接區域 256:遠端開口 258:隔離區域 260:流 262:副流 270:微小物體 280:第二培養基 290:微流體器件 300:結構/巢套 302:插座 304:電信號產生子系統 306:熱控制子系統 308:控制器 310:介面 320:印刷電路板總成(PCBA) 330:流體路徑 332:入口 334:出口 340:罩殼 350:串列埠 360:微流體器件 400:微流體器件 402:第一培養基 404:第二培養基 412:微流體線路結構 414:框架 416:微流體線路材料 420:第一光源 422:光調變子系統 424:分光器 426:置物台 428:樣本平面 430:第二光源 432:微流體線路 434:流動通道 436:生長室 440:偵測器 442:連接區域 444:隔離區域 446:隔離結構 448:二向色濾光器 450:顯微鏡 452:二向色濾光器 454:物鏡 472:近端開口 474:遠端開口 482:流體培養基流 484:副流 500A:微流體器件 500B:微流體器件 500C:微流體器件 500D:微流體器件 500E:微流體器件 502:微流體器件/儲液器 502':儲液器 504:泵連接導管 504':可透氣管道 506:轉移連接導管 508:出口連接導管 510:封閉體 512:基底 514:泵/泵組件 516:室/封閉體 518:可透氣塊 518':可透氣區段/可透氣塊 520:閥 522:轉移連接導管 524:氣體環境源 526:連接供給/連接導管 600:微流體器件 606:流體培養基入口管道 608:流體培養基出口管道 610:感測器 610':感測器 612:整合比色感測器 612':整合比色感測器 614:發光二極體(LED) 614':發光二極體(LED) 700:方法/灌注程序 800:方法 900:微流體器件 902:封閉區域 904:第一介電泳(DEP)基板 906:第二介電泳(DEP)基板 910:內表面 912:內表面 914:矽烷氧基鍵聯基團 916:調節表面 1002:細胞 1004:細胞 1006:細胞 1008:細胞 1010:細胞 1012:細胞 1014:細胞 1016:細胞 1018:細胞 1102:細胞 1202:細胞群組 1204:進一步增殖細胞群體 1206:經擴增之OKT3細胞 1302:光阱 1304:光阱 1306:釋放擴增細胞 7002:步驟 7004:步驟 8002:步驟 8004:步驟 8006:步驟 8008:步驟 D _p:副流之滲透深度 L _c1:連接區域之長度 L _c2:連接區域之長度 W _ch:通道寬度 W _con:連接區域之寬度 W _con1:近端開口之寬度 W _con2:遠端開口之寬度 100: microfluidic device 102: enclosure/lid 104: support structure/substrate 106: flow path 107: port 108: microfluidic circuit structure 109: inner surface 110: lid 114: frame 116: microfluidic circuit material 120: microfluidics line 122: microfluidic channel 124: microfluidic growth chamber/inlet 124': outlet 126: microfluidic growth chamber 128: microfluidic growth chamber 130: microfluidic growth chamber 132: micro-object trap 134: side access/flow channel / Flow area 150: System 152: Control and monitoring equipment 154: Main controller 156: Control module 158: Memory 160: Culture medium module 162: Power module 164: Imaging module 166: Tilt module 168: Other modules Group 170: Display Device 172: Input/Output Device/Control Module 178: Media Source 180: Fluid Media/Control/Monitoring Device 190: Tilt Device 192: Power Supply 194: Imaging Device 200: Microfluidic Device 202: Area/Chamber/ Fluid Medium 202': Waste Fluid Medium 202a': Stained Fluid Medium 202a'': Stained Outflow Fluid Medium 204: Bottom Electrode/Fluid Medium 206: Electrode Startup Substrate 208: Internal Surface 210: Top Electrode 212: Power Supply/Fluid Flow 214 : unirradiated dielectrophoretic (DEP) electrode area 214a: irradiated dielectrophoretic (DEP) electrode area 220: light source 222: light pattern 224: square pattern 226: controller 240: microfluidic device 242: port 244: growth Chamber 246: Growth chamber 248: Growth chamber 250: Isolation structure 252: Proximal opening 254: Connection area 256: Distal opening 258: Isolation area 260: Stream 262: Secondary stream 270: Small objects 280: Second medium 290: Micro Fluidic Device 300: Structure/Nest 302: Socket 304: Electrical Signal Generation Subsystem 306: Thermal Control Subsystem 308: Controller 310: Interface 320: Printed Circuit Board Assembly (PCBA) 330: Fluid Path 332: Inlet 334: outlet 340: housing 350: serial port 360: microfluidic device 400: microfluidic device 402: first medium 404: second medium 412: microfluidic circuit structure 414: frame 416: microfluidic circuit material 420: first light source 422: Light Modulation Subsystem 424: Optical Splitter 426: Stage 428: Sample Plane 430: Second Light Source 432: Microfluidic Circuits 434: Flow Channel 436: Growth Chamber 440: Detector 442: Connection Area 444: Isolation Area 446: Isolation structure 448: Dichroic filter 450: Microscope 452: Dichroic filter 454: Objective lens 472: Proximal opening 474: Distal opening 482: Fluid medium flow 484: Secondary flow 500A: Microfluidic device 500B: Microfluidic Device 500C: Microfluidic Device 500D: Microfluidic Device 500E: Microfluidic Device 502: Microfluidic Device/Reservoir 502': Reservoir 504: Pump Connection Conduit 504': Gas Permeable Tubing 506: Transfer Connection Conduit 508: Outlet Connection Conduit 510: Closure 512: Substrate 514: pump/pump assembly 516: chamber/closure 518: gas permeable block 518': gas permeable section/permeable block 520: valve 522: transfer connection conduit 524: gas ambient source 526: connection supply/connection conduit 600: microfluidics Device 606: Fluid Media Inlet Conduit 608: Fluid Media Outlet Conduit 610: Sensor 610': Sensor 612: Integrated Colorimetric Sensor 612': Integrated Colorimetric Sensor 614: Light Emitting Diode (LED) 614': Light Emitting Diode (LED) 700: Method/Perfusion Procedure 800: Method 900: Microfluidic Device 902: Enclosed Area 904: First Dielectrophoresis (DEP) Substrate 906: Second Dielectrophoresis (DEP) Substrate 910: inner surface 912: inner surface 914: siloxyl linking group 916: regulatory surface 1002: cell 1004: cell 1006: cell 1008: cell 1010: cell 1012: cell 1014: cell 1016: cell 1018: cell 1102: cell 1202 : cell population 1204: further proliferating cell population 1206: expanded OKT3 cells 1302: optical trap 1304: optical trap 1306: release expanded cells 7002: step 7004: step 8002: step 8004: step 8006: step 8008: step D _p : the penetration depth of the secondary flow L _c1 : the length of the connecting region L _c2 : the length of the connecting region W _ch : the width of the channel W _con : the width of the connecting region W _con1 : the width of the proximal opening W _con2 : the width of the distal opening

圖1繪示根據本發明之一些實施例之用於與一微流體器件及相關聯控制設備一起使用之一系統之一實例。圖2A及圖2B繪示根據本發明之一些實施例之一微流體器件。圖2C及圖2D繪示根據本發明之一些實施例之生長室。圖2E繪示根據本發明之一些實施例之一詳細生長室。圖2F繪示根據本發明之一實施例之一微流體器件。圖3A繪示根據本發明之一些實施例之用於與一微流體器件及相關聯控制設備一起使用之一系統之一特定實例。圖3B繪示根據本發明之一些實施例之一成像器件。圖4A至圖4C展示一微流體器件之另一實施例，其包含用於其內之一生長室之另一實例。圖5A至圖5E各表示能夠將調節培養基提供至一微流體器件以支援細胞生長、存活性、可移植性或其等之任何組合之系統組件之一實施例。圖6係具有一或多個感測器之一微流體器件之一圖示，該一或多個感測器能夠偵測進入及/或離開該微流體器件之培養基之pH。圖7係用於灌注將一流體培養基於一微流體器件中之一程序之一實施例之一實例。圖8係用於將一流體培養基灌注於一微流體器件中之一程序之另一實施例之一實例。圖9係提供進一步支援細胞生長、存活性、可移植性或其等之任何組合之一調節表面之一示意圖。圖10A至圖10E係根據本文中所描述之方法之一培養實驗之一實施例之攝影圖。圖11A係根據本文中所描述之方法之一培養實驗之另一實施例之一攝影圖，其展示將一細胞放置於一微流體器件之生長室中之前之該器件。圖11B係將一細胞放置於微流體器件之一生長室中之後之圖11A之培養實驗之一實施例之一攝影圖。圖12A至圖12C係一稍後時間點處之圖11A及圖11B之培養實驗之一實施例之攝影圖，其展示圖11B之細胞之培育期間之細胞擴增。圖13A至圖13C係一稍後時間點處之圖11A至圖11B及圖12A至圖12C之培養實驗之一實施例之攝影圖，其展示培育期結束之後之擴增細胞之輸出。圖14A及圖14B係具有至少一調節表面之一微流體器件中之另一培養實驗之一實施例之攝影圖。 Figure 1 illustrates an example of a system for use with a microfluidic device and associated control apparatus in accordance with some embodiments of the present invention. 2A and 2B illustrate a microfluidic device according to some embodiments of the present invention. 2C and 2D illustrate a growth chamber according to some embodiments of the present invention. Figure 2E illustrates a detailed growth chamber in accordance with some embodiments of the present invention. Figure 2F illustrates a microfluidic device according to one embodiment of the present invention. 3A illustrates a specific example of a system for use with a microfluidic device and associated control apparatus, according to some embodiments of the present invention. 3B illustrates an imaging device according to some embodiments of the present invention. 4A-4C show another embodiment of a microfluidic device including another example for a growth chamber therein. 5A-5E each represent one embodiment of a system component capable of providing conditioned medium to a microfluidic device to support cell growth, viability, transplantability, or any combination thereof. 6 is an illustration of a microfluidic device having one or more sensors capable of detecting the pH of the medium entering and/or leaving the microfluidic device. Figure 7 is an example of one embodiment of a procedure for perfusion based on a fluidic culture in a microfluidic device. Figure 8 is an example of another embodiment of a procedure for perfusing a fluid medium in a microfluidic device. Figure 9 is a schematic representation of a regulatory surface providing further support for cell growth, viability, transplantability, or any combination thereof. 10A-10E are photographic images of one embodiment of a culture experiment according to the methods described herein. Figure 11A is a photographic image of another embodiment of a culture experiment according to the methods described herein showing a microfluidic device prior to placing the device in the growth chamber. Figure 11B is a photograph of one embodiment of the culture experiment of Figure 11A after a cell has been placed in a growth chamber of a microfluidic device. Figures 12A-12C are photographs at a later time point of one example of the culture experiment of Figures 11A and 11B showing cell expansion during incubation of the cells of Figure 11B. Figures 13A-13C are photographs at a later time point of one example of the culture experiments of Figures 11A-11B and 12A-12C showing the output of expanded cells after the incubation period has ended. 14A and 14B are photographic images of one embodiment of another culture experiment in a microfluidic device having at least one conditioning surface.

900:微流體器件 900: Microfluidic Devices

902:封閉區域 902: Enclosed Area

904:第一介電泳(DEP)基板 904: First Dielectrophoresis (DEP) Substrate

906:第二介電泳(DEP)基板 906: Second Dielectrophoresis (DEP) Substrate

910:內表面 910: Inner Surface

912:內表面 912: Inner Surface

914:矽烷氧基鍵聯基團 914: Siloxy linking group

916:調節表面 916: Conditioning Surface

Claims

A kit for making a microfluidic device, comprising: a microfluidic device comprising one or more discrete microfluidic circuits, each configured to hold a fluid, the microfluidic circuit defined by an enclosure Defined, the enclosure includes: a base comprising a first material and having an inner surface; a microfluidic circuit structure comprising a second material and having an inner surface; and a lid comprising a third material and having an inner surface, wherein the inner surface of the substrate, the inner surface of the microfluidic circuit structure, and the inner surface of the cover each face one of the at least one growth chamber of the one or more discrete microfluidic circuits an interior; wherein the interior surfaces of the interior facing the at least one growth chamber have a monolayer covalently attached thereto, wherein the monolayer includes a bonding group and is configured to be covalently bonded to a modulating a reactive moiety _Rx of the modifier in support of cell growth, viability, portability, or any combination thereof in the at least one growth chamber, and further wherein the interior of the at least one growth chamber faces the interior of the at least one growth chamber Each linking group of the monolayer of the inner surface is the same linking group, and the conditioning modifier includes a reactive partner moiety _Rpx that is configured to react with the reactive moiety Rx of the _monolayer and part thereof for supporting cell growth, viability, transplantability or any combination thereof.

The kit of claim 1, wherein the first, second and third materials comprise different materials.

The kit of claim 1, wherein the second material comprises a polymer, and further wherein the first material is different from the polymer.

The kit of claim 1, wherein the substrate comprises one or more semiconductor substrates.

The kit of claim 1, wherein the microfluidic device further comprises a substrate having a dielectrophoresis (DEP) configuration, the substrate configured to use DEP forces to introduce at least one mammalian cell into the at least one growth chamber middle.

The kit of claim 1, wherein the cover includes one or more indium tin oxide (ITO) electrodes.

The kit of claim 1, wherein the at least one growth chamber comprises: an isolation region having a single opening; and A connecting region including a proximal opening to a flow region and a distal opening to the isolation region.

The kit of claim 1, wherein the at least one growth chamber comprises: an isolation region having a single opening; and a connecting region including a proximal opening to a flow region and one to the isolation region A distal opening, wherein a length L _con of the connecting region from the proximal opening to the distal opening is at least 1.0 times a width W _con of the connecting region at the proximal opening, and further wherein when a first When a fluid medium flows within the flow region, the connecting region is configured to allow diffusion of the first fluid medium into the isolation region while substantially not allowing flow into the isolation region.

The kit of claim 8, wherein the width W _con of the connection region at the proximal opening is about 20 microns to about 100 microns.

The kit of claim 1, wherein the one or more discrete microfluidic circuits further comprise a flow region connecting a fluid inlet and the at least one growth chamber, wherein the inner surface of the substrate, the microfluidic circuit structure The inner surface, and the inner surface of the lid each face an interior of the flow region, and wherein the first, second, and third materials each include a monolayer covalently attached thereto, wherein the monolayer includes a linking group and _is configured to covalently bond to a reactive moiety Rx that modulates the modifier for supporting cell growth, viability, transplantability or the like within the flow region in any combination , and further wherein each linking group of the monolayer of the first, second, and third materials is the same linking group.

The kit of claim 10, wherein the linking groups of the monolayer on the surface of the flow region are the same as the linking groups on the surface of the at least one growth chamber.

The kit of claim 1, wherein the linking group is attached to the substrate inner surface, the microfluidic circuit inner surface, and the lid inner surface through an oxide moiety.

The kit of claim 1, wherein the linking group is a siloxy linking group.

The kit of claim 1, wherein the monolayer facing the interior of the at least one growth chamber further comprises a linker having a backbone comprising 10 to 20 carbon atoms.

The kit of claim 1, wherein the monolayer facing the interior of the at least one growth chamber further comprises a linker of no more than 11 carbon atoms.

The kit of claim 1, wherein the modulatory modifier further comprises a linkage to the reactive partner moiety _Rpx and a linkage to the moiety for supporting cell growth, viability, transplantability, or any combination thereof, or the like , wherein the linker has 1 to 100 non-hydrogen atoms.

The kit of claim 1, wherein the portion configured to support cell growth, viability, transplantability, or any combination thereof of the modulating modifier comprises a polymer having an alkylene oxide moieties, carboxylic acid moieties, sulfonic acid moieties, phosphonic acid moieties, carbohydrate moieties, nucleotide moieties, and/or amino acid moieties.

The kit of claim 1, wherein the regulatory modifier comprises a mixture of more than one regulatory modifier, each of the regulatory modifiers comprising the same reaction pairing moiety _Rpx and is used to support cell growth, viability , portability, or any combination of more than one different part.

The kit of claim 1, wherein the reactive moiety Rx comprises an azide or an alkynyl, respectively, and the reactive partner _Rpx comprises an alkynyl or an _azide , for the The reactive partner moiety _Rpx _is covalently bonded to the reactive moiety Rx.

A method for making a microfluidic device, comprising: introducing a modulating modifier comprising a moiety for supporting cell growth, viability, transplantability, or any combination thereof, and a reaction partner moiety _Rpx into a A microfluidic circuit of a microfluidic device comprising one or more discrete microfluidic circuits, each configured to hold a fluid, the microfluidic circuit being bounded by an enclosure comprising : a substrate including a first material and having an inner surface; a microfluidic circuit structure including a second material and having an inner surface; and a cover including a third material and having an inner surface, wherein the substrate, the microfluidic circuit structure, and the inner surfaces of the cover each face inside one of at least one microfluidic circuit element of the one or more discrete microfluidic circuits, wherein the substrate, the microfluidic circuit structure , and the inner surfaces of the cover have a monolayer covalently attached thereto, wherein the monolayer includes a bonding group and is configured to covalently bond to the at least one microfluidic circuit element A reactive moiety Rx of the conditioning _modifier , and further wherein each linking group of the monolayer of the substrate, the microfluidic circuit structure, and the inner surfaces of the cover is the same linking group; and covalently bonding the reaction pairing moiety _Rpx and the reaction moiety _Rx to form a conditioning surface on the substrate of the at least one microfluidic circuit element, the microfluidic circuit structure, and the inner surfaces of the cover A monolayer, wherein the conditioning surface monolayer is configured to support cell growth, viability, transplantability, or any combination thereof, within the microfluidic device.

The method of claim 20, wherein the at least one microfluidic circuit element includes at least one growth chamber.

The method of claim 20, wherein the at least one microfluidic circuit element includes a flow region.

The method of claim 20, wherein the conditioning modifier has a structure of formula 5:

Formula 5 wherein the reactive pairing moiety _Rpx is a reactive group configured to react with the reactive moiety Rx; a moiety _is a moiety configured to support cell growth, viability, transplantability, or any combination thereof; L' is a bonder having 1 to 100 non-hydrogen atoms having any combination selected from the group consisting of silicon atoms, carbon atoms, nitrogen atoms, oxygen atoms, sulfur atoms and phosphorus atoms; m is 0 or 1.

The method of claim 23, wherein the moiety configured to support cell growth, viability, transplantability, or any combination thereof of the modulating modifier comprises a polymer having an alkylene oxide moiety , carboxylic acid moieties, sulfonic acid moieties, phosphonic acid moieties, carbohydrate moieties, nucleotide moieties, and/or amino acid moieties.

The method of claim 20, wherein the regulatory modifier comprises a mixture of more than one regulatory modifier, each of the regulatory modifiers comprising the same reaction pair moiety _Rpx and is used to support cell growth, viability, More than one different part of portability or any combination thereof.

The method of claim 20, wherein the reactive moiety Rx comprises an azide or an alkynyl group, respectively, and the reaction partner _Rpx comprises an alkynyl or an _azide group for the reaction The mating moiety _Rpx _is covalently bonded to the reactive moiety Rx.

The method of claim 20, wherein the microfluidic device comprises the substrate having the monolayer, the microfluidic circuit structure, and the inner surfaces of the lid, the monolayer comprising a bonding group and a reactive moiety R _x , wherein the method includes reacting the oxides of the surfaces with a modifying agent, thereby converting the surfaces into the modified surface having the monolayer including a linking group and a reactive moiety R _x .

The method of claim 27, wherein the modifier has a structure of formula 4: LG-(L") _j - R _x formula 4 wherein LG is a linking group; , a bonder of 1 to 100 non-hydrogen atoms of any combination of nitrogen atoms, oxygen atoms, sulfur atoms, and phosphorus atoms; Rx _is a reactive moiety; and j is 0 or 1.

The method of claim 28, wherein the linking group LG is a siloxane or phosphonic acid group.

The method of claim 28, wherein the reactive moiety Rx _is an azide, amine, bromo, thiol, active ester, succinimidyl or alkynyl moiety.

29. The method of claim 29, wherein reacting the oxides of the inner surfaces with the modifier is performed by exposing the inner surfaces to a vapor containing the modifier at a pressure below atmospheric pressure.

A method of culturing at least one biological cell in a microfluidic device, the method comprising: introducing at least one biological cell into a growth chamber of a microfluidic device, the growth chamber having a surface conditioned to support cell growth, viability, portability, or any combination thereof, wherein the microfluidic device includes a discrete microfluidic circuit configured to contain a flow of a first fluid medium, the discrete microfluidic circuit including a fluid inlet, the growth chamber, and connecting the fluid inlet and the a flow area of the growth chamber; incubating the at least one biological cell in the microfluidic device for a length of time sufficient to expand the at least one biological cell to produce a population of biological cells; and During the incubation step, the first fluid medium is perfused discontinuously through the flow area.

The method of claim 32, wherein the surface modulated to support cell growth, viability, transplantability, or any combination thereof, is covalently attached to the surface.