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CN113275053A - 试剂存储系统 - Google Patents

试剂存储系统 Download PDF

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CN113275053A
CN113275053A CN202110143518.0A CN202110143518A CN113275053A CN 113275053 A CN113275053 A CN 113275053A CN 202110143518 A CN202110143518 A CN 202110143518A CN 113275053 A CN113275053 A CN 113275053A
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D·A·阿莫雷塞
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Abstract

本申请涉及试剂存储系统。可将少量的含水样品、然后气隙和油滴抽取到移液管吸头中，以此作为长时间存储和保护所述样品免受环境污染的手段。

Description

试剂存储系统
技术领域
本公开涉及试剂和反应存储。
背景技术
许多分析技术都需要使用精确的试剂，有时需要少量的试剂。例如，在分子生物学中，通常在微流体环境中进行反应，并且必须以精确的量和比率来组合几种试剂以确保可接受的结果。此外，许多反应对污染敏感，因此必须在受控的环境中存储和操纵试剂。各种使试剂递送自动化的尝试可以消除一些人为错误，但也有其自身的缺点。
现有的自动化的试剂递送方法包含机器人流体处理器和微流体装置。计算机控制系统中自动化反应处理的出现允许进行全天候的实验。不幸的是，由于自动化反应会在深夜和周末完成，因此，每次自动化反应都可能会有数小时或数天无人看管，直到研究人员返回实验室为止。当样品等待回收时，反应体积可能会因蒸发而损失，或者反应体积会经历其它可能影响结果并破坏实验的环境质量下降。
控制这些样品的蒸发和环境质量下降至关重要。存在用于将密封件施加到含有最终样品的容器的密封系统，但是这些密封系统增加了液体处理器的成本和复杂性，并且会干扰被处理样品的获取。即使是在密封的盒中，具有大表面积的小体积仍可能会随时间蒸发。当这些样品放在敞开的容器中时，取决于操作环境(相对湿度和气流)，会更快地发生蒸发。
发明内容
本发明的系统和方法提供在常用液体反应处理部件(例如，移液管吸头)中的样品存储，这避免了其中反应体积的蒸发和环境质量下降。通过在移液管吸头中含有反应体积，可以简化处理并节省昂贵的消耗品，从而节省时间和金钱。为了存储液体样品，可以将少量其它液体吸入移液管的末端，以将样品与外部环境分离。由于移液管的功能，吸头的一端已经相对于移液管本身密封，同时液体通过相反的开口端吸入。通过将密封液吸入开口端，随后可以将样品体积完全密封在移液管吸头内，从而避免蒸发和其它形式的环境质量下降。密封液可以不与样品混溶，以防止混合。例如，在样品是含水的情况下，密封液可以是油，诸如矿物油。密封液可以是惰性的或不与样品反应，以避免对所存储的样品的反应产生任何影响。移液管吸头可以是用于手动移液管或用于自动化系统的由聚合物(例如，聚丙烯、具有石墨的聚丙烯)制成的一次性吸头。移液管吸头也可以是由涂覆有含氟聚合物的不锈钢制成的套管，也可以不是用于自动化处理系统的套管。
在某些实施例中，可以在吸入密封液之前将少量空气吸入移液管吸头，以在密封液和样品体积之间提供分离。样品与密封液之间形成的气隙可以防止密封液迁移到样品体积上方或与样品体积交换位置，而所述迁移将有效地破坏密封。在一些实施例中，密封液可以具有比样品更大的密度，并且移液管可以以直立的方式存储，使得吸头开口在最低点，并且更稠密的密封液保留在样品和开口之间。每当可能存储试剂或反应体积时，诸如在反应完成时或在反应停止点时，都可以使用本发明的系统和方法。在某些实施例中，根据本发明的方法，可以在密封的移液管吸头中执行实验中的温育或其它较长时间的步骤。
本发明的方法可以手动执行或在自动化流体处理机中执行。在某些实施例中，自动化流体处理装置可以由包含非暂时性存储器和处理器的计算机控制。计算机可操作以执行所述方法的步骤，从而在反应结束时存储样品。计算机可以在接收到用于暂停反应的用户命令时启动本文所述的方法的密封步骤。可以经由用户界面接收此类命令。在某些实施例中，计算机可以被编程为在反应完成时或在等待额外的用户输入时自动密封移液管吸头，直到用户准备重新开始反应或取回样品为止。自动化机器可以包括密封液的储器，以执行密封步骤。
本发明的各方面包含样品存储方法。所述方法可包含：在附接到移液管的移液管吸头中提供样品；以及使用移液管通过移液管吸头的开口抽取空气，以在样品和开口之间形成气隙。然后，方法可包含使用移液管将流体吸入开口，以密封开口并防止样品蒸发。
流体可不与样品混溶。在各种实施例中，样品可是含水的，并且流体可是油。流体可包括矿物油。在某些实施例中，流体可以是蜡；并且方法可以包含将移液管吸头在低于蜡的熔点的温度下存储，并且将蜡加热到高于其熔点以从吸头的存储中移出样品。在某些实施例中，气隙可以将流体与样品分离。方法可以包含：将样品在没有蒸发的情况下存储在移液管吸头中至少一小时、至少6小时、至少12小时、至少24小时、至少2天或至少3天。
在一些实施例中，移液管可以是自动化处理系统的一部分。自动化处理系统可以包含与处理器通信的非暂时性存储器，所述处理器可操作以使用移液管执行反应步骤。自动化处理系统可操作以在反应完成时执行抽取步骤。自动化处理系统可操作以在接收到暂停反应的用户输入时执行抽取。
在某些方面，本发明的系统可以包含自动化处理系统，所述自动化处理系统包含计算机和移液管并且可操作以执行本文所述的方法。
附图说明
图1示出了根据某些实施例的移液管吸头存储。
图2示出了根据某些实施例的具有缓冲物的移液管吸头存储。
图3图示了本发明的方法的步骤。
图4图示了用于具有缓冲物的样品存储的方法的步骤。
具体实施方式
本发明的系统和方法提供了试剂和样品在反应期间或之后的流线型存储。通过将所述体积存储在原本存在且用于反应处理的移液管吸头中，可以避免使用复杂的密封系统和过多的消耗品。由于样品被密封在移液管吸头内，因此可以避免由样品蒸发或环境质量下降引起的问题。因此，在周末或其它非工作时间到达停止点的自动化过程中，系统可以将样品安全地存储在移液管吸头中数小时或数天，直到研究人员或其它用户可以看管其为止。
移液管吸头避免了由于较小的横截面面积通过移液管吸头的开口暴露于环境导致的环境暴露所引起的大量蒸发和质量下降问题。即便如此，本发明的系统和方法包含：将密封流体吸入移液管吸头开口，以在样品或反应体积与外部环境之间形成缓冲物。
图1示出了根据某些实施例，样品体积105存储在移液管吸头101中。移液管吸头101包括开口103，响应于移液管(未示出)中的压力或体积的变化，流体可以通过所述开口被吸入移液管吸头101。移液管吸头101可以是包含被密封到移液管(未示出)的较大开口的一次性吸头，诸如可从帝肯贸易有限公司(Tecan Trading,Ltd.)(瑞士)商购的那些。在某些实施例中，移液管吸头101可以被集成到移液管中，所述移液管包括例如用于将流体吸入吸头101并从所述吸头排出流体的可变形球状物。移液管和移液管吸头101可以是自动化流体处置系统的一部分，所述系统诸如可以从帝肯贸易有限公司(瑞士)获得的Fluent andFreedom EVO平台。
当需要存储样品体积105时，在将密封液109吸入移液管吸头101之前，可以使用移液管将空气吸入移液管吸头101以形成气隙107。密封液109可与样品体积105不混溶，以防止其接触时发生混合。气隙107可以有助于防止任何此类接触，并且有助于维持样品体积105和密封液109相对于开口103的位置，从而维持样品体积的密封环境以防止蒸发或环境质量下降。
在优选的实施例中，样品体积105是含水的，诸如已经准备好用于测序的DNA文库。在此类实施例中，密封液109可以是油，诸如矿物油。实例性矿物油包含可从赛默飞世尔(ThermoFisher)(马萨诸塞州沃尔瑟姆)获得的J6292、BP26291和O121-1以及可从西格玛奥德里奇(Sigma-Aldrich)(密苏里州圣路易斯)获得的M5904生物试剂和M8662PCR试剂。在优选的实施例中，密封液109可以是低挥发性流体，所述流体在室温下不蒸发并且因此是无味的。
在一些实施例中，密封液109可以是蜡，所述蜡以液态被吸入移液管吸头101。然后可以将移液管吸头101存放在低于蜡熔点的温度下，并且然后可以将密封蜡加热到高于其熔点，以去除样品体积105。密封液109优选地对样品体积105是惰性的或不反应的，使得两者之间的任何偶然接触都将不会影响反应结果。为了促进密封液109在样品体积105和开口103之间的期望定位，可以操纵两种流体的相对密度和移液管吸头101的存储位置。例如，移液管吸头101可以在开口103朝下的情况下进行存储，并且可以选择密封液109使其密度大于样品体积105。
图2示出了根据某些实施例，样品体积105和缓冲体积108存储在移液管吸头101中。移液管吸头101包括开口103，响应于移液管(未示出)中的压力或体积的变化，流体可以以与关于图1所述的方式类似的方式通过所述开口被吸入移液管吸头101。然而，额外的缓冲体积108被吸入移液管吸头101，并通过两个气隙(106和107)与样品体积105和密封液109隔开。在某些实施例中，缓冲体积108可以是溶剂，并且可以具有与样品体积105相似的组成(例如，相似的pH和离子强度)。
图3图示了本发明的实例性方法201的步骤。为了启动样品体积的移液管吸头存储，首先使用移液管通过吸头的开口将样品吸入移液管吸头203。如所指出的，移液管可以手动操作，或者可以是自动化系统的一部分。随后，将空气吸入移液管吸头205，以在样品和移液管吸头的开口之间形成气隙。然后将密封液吸入移液管吸头207中。在手动操作的移液管中，应选择足够的体积以容纳要存储的样品体积、气隙和密封液。
如图4所示，在将样品吸入移液管吸头203和将空气吸入移液管吸头205之间，可以在206抽取缓冲物。如图2所示，可以在204在样品和缓冲物之间将额外的空气体积吸入以获得样品、空气间隙、缓冲物、气隙、密封液布置。
自动化系统(诸如可从帝肯贸易有限公司获得的那些)可以使用移液管在储器和反应容器之间自动分配精确体积的样品和试剂。因此，此类系统已经包含移液管吸头并且可操作以操纵它们用于流体处理的目的。此类系统用于执行各种反应，包含基因组应用和基于细胞的测定。可以根据其中包含的反应的特定要求将移液管吸头内存储编程进入任何测定。例如，在较长的温育期间，如本文所述的移液管吸头存储可用于避免在温育期间蒸发或样品质量下降。可以将样品存储步骤编程进入自动化系统，作为完成测定或其它反应后的默认最后一步。因此，如果自动化反应在周末或下班后完成，则将安全地存储所述反应，直到用户可以取回样品进行分析、长期存储或其它后续处理为止。
在测定性质允许的情况下，自动化系统可以提供经由用户界面暂停测定的选项，使得用户可以表示希望中止反应，并且在达到测定的下一个机会时(例如，在反应对时间不敏感的情况下)，可以应用本文所述的移液管吸头内储存方法。在某些实施例中，可以经由自动化系统中的加热器或冰箱来操纵存储在移液管吸头中样品的温度，以帮助温育或促进所存储的样品的稳定性。此类系统可以包括可以在其中执行移液操作以及潜在地执行样品存储的盒。在某些实施例中，系统可包含给盒内部消毒以避免样品污染的装置。例如，系统可操作以通过各种已知方法将消毒气体释放到盒中和/或产生消毒气体，诸如臭氧气体。可以通过任何方法来完成消毒，包含通过应用消毒能量，诸如光(例如紫外线)或热量来完成。
可以修改本发明的系统和方法，使得替代将密封液吸入移液管吸头中，移液管吸头的开口可以浸没在流体储器中以有效地密封移液管吸头并防止其内容物蒸发。储器可以含有以上讨论的任何流体作为潜在的密封液，包含各种矿物油或者甚至蜡。通过浸没来密封移液管吸头也与如上讨论的气隙和缓冲物的使用兼容，并且可以设想此类实施例。
在各种实施例中，当需要取回样品体积时，可以释放密封液和样品。在存储期间，自动化系统可以精确管理和记录移液管吸头中吸入的空气和密封液的体积，然后可以使用此信息将用过的准确量的密封液丢弃到废物容器中，然后再分配样品以进行进一步处理或分析。气隙可以提供在排空密封液时有用的缓冲物，而不会意外丢弃任何样品体积。
在某些实施例中，缓冲物和密封液可以在单个分配动作中被丢弃。当缓冲物从吸头分配时，缓冲物可用来洗去痕量的密封液，从而避免密封液潜在污染样品。
使用本文所述方法存储在移液管吸头中的样品可以在无可测量蒸发的情况下保存超过1小时、2小时、6小时、12小时、24小时、2天、3天、4天或5天。
如本领域技术人员将认识到的必要或最适合本发明的系统和方法，本发明的系统和方法可以包含一或多个服务器和/或计算装置，其可以包含一或多个经由总线相互通信的处理器(例如，中央处理器(CPU)、图形处理器(GPU)等)、计算机可读存储装置(例如，主存储器、静态存储器等)或其组合。计算机系统可以集成到本发明的自动化处理系统中或与所述自动化处理系统通信，并且可操作以控制其中的一或多个移液管并执行上述方法。
处理器可以包含本领域中已知的任何合适的处理器，诸如由英特尔(Intel)(加利福尼亚州圣克拉拉)以商标XEON E7出售的处理器或由超威半导体公司(AMD)(加利福尼亚州森尼维耳)以商标OPTERON 6200出售的处理器。
存储器优选地包含至少一个有形的非暂时性媒体，所述媒体能够存储：一或多个的指令集合，所述指令可执行以使系统执行本文所述的功能(例如，体现本文中出现的任何方法或功能的软件)；数据；或两者。虽然在实例性实施例中计算机可读存储装置可以是单个媒体，但术语“计算机可读存储装置”应被认为包含存储指令或数据的单个媒体或多个媒体(例如，集中式或分布式数据库，和/或相关联的高速缓冲存储器和服务器)。因此，术语“计算机可读存储装置”应被认为包含但不限于固态存储器(例如，用户识别模块(SIM)卡、安全数字卡(SD卡)、微型SD卡或固态驱动器(SSD))、光学和磁性媒体、硬盘驱动器、磁盘驱动器以及任何其它有形存储媒体。
任何适合的服务都可以用于存储，诸如例如亚马逊网络服务(Amazon WebServices)、服务器的内存、云存储、另一台服务器或其它计算机可读存储装置。云存储可以指一种数据存储方案，其中数据存储在逻辑池中，并且物理存储可以跨越多个服务器和多个位置。存储装置可以由托管公司拥有和管理。优选地，存储装置用于根据需要存储指令以执行和支持本文所述的操作。
根据本发明的输入/输出装置可以包含以下中的一或多个：视频显示单元(例如，液晶显示器(LCD)或阴极射线管(CRT)监视器)、字母数字输入装置(例如，键盘)、光标控制装置(例如，鼠标或跟踪垫)、磁盘驱动单元、信号生成装置(例如，扬声器)、触摸屏、按钮、加速计、麦克风、蜂窝式射频天线、网络接口装置(其例如可以是网络接口卡(NIC)、Wi-Fi卡或蜂窝式调制解调器)或其任何组合。输入/输出装置(诸如用户界面)可用于提供有关反应状态的信息，并接收包含反应保持的指令，计算机可将所述指令解释为执行本文所述移液管上存储方法的指令。
本领域技术人员将认识到，可以采用任何合适的开发环境或编程语言来允许本文所述的用于本发明的各种系统和方法的可操作性。例如，可以使用Perl、Python、C++、C#、Java、JavaScript、Visual Basic、Ruby on Rails、Groovy和Grails或任何其它合适的工具来实现本文的系统和方法。
通过引用并入
在整个本公开中，已经引用了其它文件，诸如专利、专利申请、专利出版物、期刊、书籍、论文、网页内容。出于所有目的，所有此类文件于此通过引用以其整体并入本文。
等效物
根据本文的全部内容，包含对本文引用的科学文献和专利文献的引用，除了本文显示和描述的那些之外，本发明的各种修饰及其许多其它实施例对于本领域技术人员而言也将变得显而易见。本文主题含有适用于将本发明实践于其各种实施例及其等效物中的重要信息、例证和指导。

Claims (30)

1.一种样品存储方法，包括：
在附接到移液管的移液管吸头中提供样品；
使用所述移液管通过所述移液管吸头的开口抽取空气，以在所述样品和所述开口之间形成气隙；
使用所述移液管将流体吸入所述开口，以密封所述开口并防止所述样品蒸发。
2.根据权利要求1所述的方法，其中所述流体与所述样品不混溶。
3.根据权利要求1所述的方法，其中所述流体在室温下不蒸发。
4.根据权利要求1所述的方法，其中所述流体在室温下是固体。
5.根据权利要求2所述的方法，其中所述样品是含水的，并且所述流体是油。
6.根据权利要求5所述的方法，其中所述流体包括矿物油。
7.根据权利要求1所述的方法，其中所述气隙将所述流体与所述样品分离。
8.根据权利要求1所述的方法，进一步包括将所述样品在没有蒸发的情况下存储在所述移液管吸头中至少一小时。
9.根据权利要求8所述的方法，进一步包括将所述样品在没有蒸发的情况下存储在所述移液管吸头中至少6小时。
10.根据权利要求9所述的方法，进一步包括将所述样品在没有蒸发的情况下存储在所述移液管吸头中至少12小时。
11.根据权利要求10所述的方法，进一步包括将所述样品在没有蒸发的情况下存储在所述移液管吸头中至少24小时。
12.根据权利要求11所述的方法，进一步包括将所述样品在没有蒸发的情况下存储在所述移液管吸头中至少2天。
13.根据权利要求12所述的方法，进一步包括将所述样品在没有蒸发的情况下存储在所述移液管吸头中至少3天。
14.根据权利要求1所述的方法，其中所述移液管是自动化处理系统的一部分。
15.根据权利要求14所述的方法，其中所述自动化处理系统包括与处理器通信的非暂时性存储器，所述处理器可操作以使用所述移液管执行反应步骤。
16.根据权利要求15所述的方法，其中所述自动化处理系统可操作以在所述反应完成时执行抽取步骤。
17.根据权利要求15所述的方法，其中所述自动化处理系统可操作以在接收到暂停所述反应的用户输入时执行所述抽取步骤。
18.根据权利要求15所述的方法，其中所述自动化处理系统可操作以在接收到用于分配所存储的样品的用户输入时分配所述样品。
19.根据权利要求1所述的方法，进一步包括：
将所述带有密封开口的移液管吸头存放在盒中；以及
给所述盒消毒。
20.根据权利要求19所述的方法，其中使用消毒气体进行消毒。
21.根据权利要求20所述的方法，其中所述消毒气体是臭氧气体。
22.根据权利要求19所述的方法，其中使用消毒能量源进行消毒。
23.根据权利要求22所述的方法，其中所述能量是紫外线。
24.根据权利要求23所述的方法，其中所述移液管吸头阻挡紫外线。
25.根据权利要求22所述的方法，其中所述能量是热能。
26.一种样品存储方法，包括：
在附接到移液管的移液管吸头中提供样品；
使用所述移液管通过所述移液管吸头的开口抽取空气，以在所述样品和所述开口之间形成气隙；
将所述开口浸没在含有流体的储器中，以密封所述开口并防止所述样品蒸发。
27.根据权利要求26所述的样品存储方法，其中所述流体与所述样品不混溶。
28.根据权利要求27所述的样品存储方法，其中所述样品是含水的，并且所述流体是油。
29.根据权利要求28所述的样品存储方法，其中所述流体包括矿物油。
30.根据权利要求26所述的样品存储方法，其中所述气隙将所述流体与所述样品分离。
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