JP3923856B2 - Manufacturing method of microarray chip - Google Patents

Description

【００８８】
ただし、ｎは２以上の整数であり、Ｒ^１，Ｒ^２はそれぞれ炭素数１〜１０の置換もしくは非置換のアルキル、アルケニル、アリールあるいはシアノアルキル基であり、モル比で全体の４０％以下がビニル、フェニル、ハロゲン化フェニルである。また、Ｒ^１、Ｒ^２がメチル基のものが表面エネルギーが最も小さくなるので好ましく、モル比でメチル基が６０％以上であることが好ましい。また、鎖末端もしくは側鎖には、分子鎖中に少なくとも１個以上の水酸基等の反応性基を有する。
【００８９】
また、上記のオルガノポリシロキサンとともに、ジメチルポリシロキサンのような架橋反応をしない安定なオルガノシリコーン化合物を混合してもよい。
【００９０】
本態様における濡れ性変化層には、さらに界面活性剤を含有させることができる。具体的には、日光ケミカルズ（株）製ＮＩＫＫＯＬ ＢＬ、ＢＣ、ＢＯ、ＢＢの各シリーズ等の炭化水素系、デュポン社製ＺＯＮＹＬ ＦＳＮ、ＦＳＯ、旭硝子（株）製サーフロンＳ−１４１、１４５、大日本インキ化学工業（株）製メガファックＦ−１４１、１４４、ネオス（株）製フタージェントＦ−２００、Ｆ２５１、ダイキン工業（株）製ユニダインＤＳ−４０１、４０２、スリーエム（株）製フロラードＦＣ−１７０、１７６等のフッ素系あるいはシリコーン系の非イオン界面活性剤を挙げることかでき、また、カチオン系界面活性剤、アニオン系界面活性剤、両性界面活性剤を用いることもできる。
【００９１】
また、濡れ性変化層には上記の界面活性剤の他にも、ポリビニルアルコール、不飽和ポリエステル、アクリル樹脂、ポリエチレン、ジアリルフタレート、エチレンプロピレンジエンモノマー、エポキシ樹脂、フェノール樹脂、ポリウレタン、メラミン樹脂、ポリカーボネート、ポリ塩化ビニル、ポリアミド、ポリイミド、スチレンブタジエンゴム、クロロプレンゴム、ポリプロピレン、ポリブチレン、ポリスチレン、ポリ酢酸ビニル、ポリエステル、ポリブタジエン、ポリベンズイミダゾール、ポリアクリルニトリル、エピクロルヒドリン、ポリサルファイド、ポリイソプレン等のオリゴマー、ポリマー等を含有させることができる。
【００９２】
このような濡れ性変化層は、上述した成分を必要に応じて他の添加剤とともに溶剤中に分散して塗布液を調製し、この塗布液を基材上に塗布することにより形成することができる。使用する溶剤としては、エタノール、イソプロパノール等のアルコール系の有機溶剤が好ましい。塗布はスピンコート、スプレーコート、ディッブコート、ロールコート、ビードコート等の公知の塗布方法により行うことができる。また、紫外線硬化型の成分を含有している場合、紫外線を照射して硬化処理を行うことにより濡れ性変化層を形成することかできる。
【００９３】
本態様において、この濡れ性変化層の厚みは、光触媒による濡れ性の変化速度等の関係より、０．００１μｍから１μｍであることが好ましく、特に好ましくは０．０１〜０．１μｍの範囲内である。
【００９４】
本態様において上述した成分の濡れ性変化層を用いることにより、接触する光触媒処理層中の光触媒の作用により、上記成分の一部である有機基や添加剤の酸化、分解等の作用を用いて、エネルギー照射部の濡れ性を変化させて親水性とし、未照射部との濡れ性に大きな差を生じさせることができる。よって、固定化層形成用塗工液等との受容性（親水性）および反撥性（撥水性）を高めることによって、品質の良好でかつコスト的にも有利なマイクロアレイチップを得ることができる。
【００９５】
なお、本発明に用いられる濡れ性変化層は、上述したように光触媒の作用により濡れ性の変化する層であれば特に限定されるものではないが、特に、光触媒を含まない層であることが好ましい。このように濡れ性変化層内に光触媒が含まれなければ、マイクロアレイチップとして用いた場合に、光触媒の半永久的な作用がマイクロアレイチップに及ぶことが無く、長期間に渡り問題なく使用することが可能だからである。
【００９６】
また、本態様におけるマイクロアレイチップ用基材は、通常基材上に濡れ性変化層が形成されてなるものであるが、本態様においては、この濡れ性変化層が自己支持性を有する材料で形成されている場合には、基材を含まないものであってもよい。
【００９７】
このような自己支持性を有する濡れ性変化層の材料として、具体的には、光触媒処理層をその表面に接触させてエネルギーを照射することにより、その後塗布する固定化層形成用塗工液が有する表面張力と同等の表面張力の液体に対する接触角が、少なくとも１°以上、好ましくは５°以上、特に１０°以上変化する材料を挙げることができる。
【００９８】
また、この濡れ性変化層は、照射されるエネルギーを透過することができる材料で形成されていることが必要である。
【００９９】
このような材料としては、例えば、ポリエチレン、ポリカーボネート、ポリプロピレン、ポリスチレン、ポリエステル、ポリビニルフロライド、アセタール樹脂、ナイロン、ABS、PTFE、メタクリル樹脂、フェノール樹脂、ポリ弗化ビニリデン、ポリオキシメチレン、ポリビニルアルコール、ポリ塩化ビニル、ポリエチレンテレフタレート、シリコーン等を挙げることができる。
【０１００】
（濡れ性変化層上の親水性領域のパターン）
上記濡れ性変化層上における親水性領域のパターンの配置は、標識に由来する信号を検出する際に、親水性領域の位置（すなわち、その上に形成される固定化層および特異性生体高分子の位置）が確認可能である配置である限り、特に限定されるものではなく、例えば、分析目的に応じて、あるいは、標識に由来する信号を検出するのに用いる分析装置に応じて、適宜決定することができる。
【０１０１】
このような親水性領域のパターンは、検出が容易である点に鑑みれば整列して配置されていることが好ましい。また、親水性領域の複数部分を、所定の間隔をあけて線状に一列に配置し、さらに、そのような列を相互に平行に複数列配置したパターンであることがより好ましい。このように親水性領域を配置したパターンであれば、特異性生体高分子と結合する高分子が有する標識に由来する信号の検出工程を自動化することが容易である。
【０１０２】
上記濡れ性変化層上の親水性領域の間隔は、特異性生体高分子と結合する高分子が有する標識標識に由来する信号を検出する際に、隣接する親水性領域の影響を受けない程度に分離している限り、特に限定されるものではなく、例えば、隣接する親水性領域の縁部間の最短距離が、０．０１μｍ〜１ｃｍ、好ましくは０．０５μｍ〜５ｍｍの範囲内であることが好ましい。また、濡れ性変化層上の親水性領域の数も、特に限定されるものではなく、基材の大きさにも依存するが、例えば、数個〜数万個（一般には数十〜１乃至２万個）としてもよい。
【０１０３】
このような濡れ性変化層のパターンの形成方法は、本発明においては、光触媒を含有する光触媒処理層を、濡れ性変化層を向かい合うように配置した状態で、フォトマスクを用いたエネルギーのパターン照射により行うことが可能である。また、後述するように光触媒処理層側基板に光触媒処理層側遮光部が形成されているような場合は、エネルギーの全面照射を行なうようにしてもよい。
【０１０４】
（２）基材
次いで、基材について説明する。本態様においては、上述した濡れ性変化層が自己支持性に乏しい場合には、基材上に濡れ性変化層が形成されているマイクロアレイチップ用基材であってもよい。
【０１０５】
このような基材に用いられる材料としては、核酸ハイブリダイゼーションアッセイ又は免疫学的アッセイにおいて一般的に用いられている水又は有機溶媒に実質的に不溶性である有機材料又は無機材料、例えば、シリコン、ガラス、磁性金属若しくは非磁性金属、又はプラスチック等を挙げることができる。特に、シリコン、各種プラスチック類（例えば、ポリカーボネート、ポリスチレン、又はポリプロピレン等）、石英、又はガラス等を用いることが好ましい。このような基材の形状としては、少なくとも１つの平坦表面を有するものである限り、その形状は特に限定されるものではないが、プレート状又はフィルム状であることが好ましいといえる。
【０１０６】
Ｂ．パターン形成工程
次に、本態様におけるパターン形成工程について説明する。本工程は、基板上に少なくとも光触媒を含有する光触媒処理層が形成されてなる光触媒処理層側基板を用い、前記マイクロアレイチップ用基材および前記光触媒処理層側基板を、前記光触媒処理層および前記濡れ性変化層が向かい合うように、かつ、２００μｍ以下の間隙をおいて配置した後、所定の方向からエネルギーを照射することにより、前記濡れ性変化層表面に濡れ性の違いによるパターンを形成する工程である。以下、このような本工程で用いる光触媒処理層側基板およびエネルギー照射について説明する。
【０１０７】
なお、本工程は、本態様の他に、後述する第２実施態様においても行われる工程である。そこで、以下の本工程の説明については、本発明の両方の態様に対応させるため、上述した濡れ性変化層および後述する分解除去層を総称して特性変化層と記載する場合がある。
【０１０８】
（１）光触媒処理層側基板
本工程においては、後述するエネルギー照射において、光触媒処理層側基板を用いることを特徴とする。この光触媒処理層側基板は、少なくとも光触媒処理層と基板とを有するものであり、通常は基板上に所定の方法で形成された薄膜状の光触媒処理層が形成されてなるものである。また、この光触媒処理層側基板には、パターン状に形成された光触媒処理層側遮光部が形成されたものも用いることができる。
【０１０９】
ａ．光触媒処理層
本発明に用いられる光触媒処理層は、光触媒処理層中の光触媒が、特性変化層の特性を変化させるような構成であれば、特に限定されるものではなく、光触媒とバインダとから構成されているものであってもよいし、光触媒単体で成膜されたものであってもよい。また、その表面の濡れ性は特に親水性であっても撥水性であってもよい。
【０１１０】
本発明において用いられる光触媒処理層は、例えば上記図１（ａ）等に示すように、基板１上に全面に形成されたものであってもよいが、例えば図８に示すように、基板１上に光触媒処理層２がパターン上に形成されたものであってもよい。
【０１１１】
このように光触媒処理層をパターン状に形成することにより、後述するエネルギー照射において説明するように、光触媒処理層を特性変化層と所定の間隔をおいて配置させてエネルギーを照射する際に、フォトマスク等を用いるパターン照射をする必要がなく、全面に照射することにより、特性変化層上に特性の変化したパターンを形成することができる。
【０１１２】
この光触媒処理層のパターニング方法は、特に限定されるものではないが、例えばフォトリソグラフィー法等により行うことが可能である。
【０１１３】
また、実際に光触媒処理層に面する特性変化層上の部分のみの特性が変化するものであるので、エネルギーの照射方向は上記光触媒処理層と特性変化層とが面する部分にエネルギーが照射されるものであれば、いかなる方向から照射されてもよく、さらには、照射されるエネルギーも特に平行光等の平行なものに限定されないという利点を有するものとなる。
【０１１４】
このように光触媒処理層における、後述するような二酸化チタンに代表される光触媒の作用機構は、必ずしも明確なものではないが、光の照射によって生成したキャリアが、近傍の化合物との直接反応、あるいは、酸素、水の存在下で生じた活性酸素種によって、有機物の化学構造に変化を及ぼすものと考えられている。本発明においては、このキャリアが光触媒処理層近傍に配置される特性変化層中の化合物に作用を及ぼすものであると思われる。
【０１１５】
本発明で使用する光触媒としては、光半導体として知られる例えば二酸化チタン（ＴｉＯ_２）、酸化亜鉛（ＺｎＯ）、酸化スズ（ＳｎＯ_２）、チタン酸ストロンチウム（ＳｒＴｉＯ_３）、酸化タングステン（ＷＯ_３）、酸化ビスマス（Ｂｉ_２Ｏ_３）、および酸化鉄（Ｆｅ_２Ｏ_３）を挙げることができ、これらから選択して１種または２種以上を混合して用いることができる。
【０１１６】
本発明においては、特に二酸化チタンが、バンドギャップエネルギーが高く、化学的に安定で毒性もなく、入手も容易であることから好適に使用される。二酸化チタンには、アナターゼ型とルチル型があり本発明ではいずれも使用することができるが、アナターゼ型の二酸化チタンが好ましい。アナターゼ型二酸化チタンは励起波長が３８０ｎｍ以下にある。
【０１１７】
このようなアナターゼ型二酸化チタンとしては、例えば、塩酸解膠型のアナターゼ型チタニアゾル（石原産業（株）製ＳＴＳ−０２（平均粒径７ｎｍ）、石原産業（株）製ＳＴ−Ｋ０１）、硝酸解膠型のアナターゼ型チタニアゾル（日産化学（株）製ＴＡ−１５（平均粒径１２ｎｍ））等を挙げることができる。
【０１１８】
光触媒の粒径は小さいほど光触媒反応が効果的に起こるので好ましく、平均粒径か５０ｎｍ以下が好ましく、２０ｎｍ以下の光触媒を使用するのが特に好ましい。
【０１１９】
本発明における光触媒処理層は、上述したように光触媒単独で形成されたものであってもよく、またバインダと混合して形成されたものであってもよい。
【０１２０】
光触媒のみからなる光触媒処理層の場合は、特性変化層上の特性の変化に対する効率が向上し、処理時間の短縮化等のコスト面で有利である。一方、光触媒とバインダとからなる光触媒処理層の場合は、光触媒処理層の形成が容易であるという利点を有する。
【０１２１】
光触媒のみからなる光触媒処理層の形成方法としては、例えば、スパッタリング法、ＣＶＤ法、真空蒸着法等の真空製膜法を用いる方法を挙げることができる。真空製膜法により光触媒処理層を形成することにより、均一な膜でかつ光触媒のみを含有する光触媒処理層とすることが可能であり、これにより特性変化層上の特性を均一に変化させることが可能であり、かつ光触媒のみからなることから、バインダを用いる場合と比較して効率的に特性変化層上の特性を変化させることが可能となる。
【０１２２】
また、光触媒のみからなる光触媒処理層の形成方法としては、例えば光触媒が二酸化チタンの場合は、基板上に無定形チタニアを形成し、次いで焼成により結晶性チタニアに相変化させる方法等が挙げられる。ここで用いられる無定形チタニアとしては、例えば四塩化チタン、硫酸チタン等のチタンの無機塩の加水分解、脱水縮合、テトラエトキシチタン、テトライソプロポキシチタン、テトラ−ｎ−プロポキシチタン、テトラブトキシチタン、テトラメトキシチタン等の有機チタン化合物を酸存在下において加水分解、脱水縮合によって得ることができる。次いで、４００℃〜５００℃における焼成によってアナターゼ型チタニアに変性し、６００℃〜７００℃の焼成によってルチル型チタニアに変性することができる。
【０１２３】
また、バインダを用いる場合は、バインダの主骨格が上記の光触媒の光励起により分解されないような高い結合エネルギーを有するものが好ましく、例えばオルガノポリシロキサン等を挙げることができる。
【０１２４】
このようにオルガノポリシロキサンをバインダとして用いた場合は、上記光触媒処理層は、光触媒とバインダであるオルガノポリシロキサンとを必要に応じて他の添加剤とともに溶剤中に分散して塗布液を調製し、この塗布液を基板上に塗布することにより形成することができる。使用する溶剤としては、エタノール、イソプロパノール等のアルコール系の有機溶剤が好ましい。塗布はスピンコート、スプレーコート、ディッブコート、ロールコート、ビードコート等の公知の塗布方法により行うことができる。バインダとして紫外線硬化型の成分を含有している場合、紫外線を照射して硬化処理を行うことにより光触媒処理層を形成することかできる。
【０１２５】
また、バインダとして無定形シリカ前駆体を用いることができる。この無定形シリカ前駆体は、一般式ＳｉＸ_４で表され、Ｘはハロゲン、メトキシ基、エトキシ基、またはアセチル基等であるケイ素化合物、それらの加水分解物であるシラノール、または平均分子量３０００以下のポリシロキサンが好ましい。
【０１２６】
具体的には、テトラエトキシシラン、テトライソプロポキシシラン、テトラ−ｎ−プロポキシシラン、テトラブトキシシラン、テトラメトキシシラン等が挙げられる。また、この場合には、無定形シリカの前駆体と光触媒の粒子とを非水性溶媒中に均一に分散させ、基板上に空気中の水分により加水分解させてシラノールを形成させた後、常温で脱水縮重合することにより光触媒処理層を形成できる。シラノールの脱水縮重合を１００℃以上で行えば、シラノールの重合度が増し、膜表面の強度を向上できる。また、これらの結着剤は、単独あるいは２種以上を混合して用いることができる。
【０１２７】
バインダを用いた場合の光触媒処理層中の光触媒の含有量は、５〜６０重量％、好ましくは２０〜４０重量％の範囲で設定することができる。また、光触媒処理層の厚みは、０．０５〜１０μｍの範囲内が好ましい。
【０１２８】
また、光触媒処理層には上記の光触媒、バインダの他に、界面活性剤および添加剤を含有させることができる。これら界面活性剤および添加剤については、上述した「（１）濡れ性変化層」と同様であるので、ここでの説明は省略する。
【０１２９】
ｂ．基板
本発明においては、図１に示すように、光触媒処理層側基板３は、少なくとも基板１とこの基板１上に形成された光触媒処理層２とを有するものである。
【０１３０】
この際、用いられる基板を構成する材料は、後述するエネルギー照射の方向等により適宜選択される。
【０１３１】
すなわち、例えばマイクロアレイチップ用基材が不透明なものを基材として用いる場合においては、エネルギー照射方向は必然的に光触媒処理層側基板側からとなり、図１（ｂ）に示すように、フォトマスク７を光触媒処理層側基板３側に配置して、エネルギー照射をする必要がある。また、後述するように光触媒処理層側基板に光触媒処理層側遮光部を予め所定のパターンで形成しておき、この光触媒処理層側遮光部を用いてパターンを形成する場合においても、光触媒処理層側基板側からエネルギーを照射する必要がある。このような場合、基板は透明性を有するものであることが必要となる。
【０１３２】
一方、マイクロアレイチップ用基材が透明である場合であれば、このマイクロアレイチップ用基材側にフォトマスクを配置してエネルギーを照射することも可能である。このような場合、基板には特に透明性は要求されない。
【０１３３】
また本発明に用いられる基板は、可撓性を有するもの、例えば樹脂製フィルム等であってもよいし、可撓性を有さないもの、例えばガラス基板等であってもよい。これは、後述するエネルギーの照射方法により適宜選択されるものである。
【０１３４】
このように、本発明における光触媒処理層側基板に用いられる基板は特にその材料を限定されるものではないが、本発明においては、この光触媒処理層側基板は、繰り返し用いられるものであることから、所定の強度を有し、かつその表面が光触媒処理層との密着性が良好である材料が好適に用いられる。
【０１３５】
具体的には、ガラス、セラミック、金属、プラスチック等を挙げることができる。
【０１３６】
なお、基板表面と光触媒処理層との密着性を向上させるために、基板上にアンカー層を形成するようにしてもよい。このようなアンカー層としては、例えば、シラン系、チタン系のカップリング剤等を挙げることができる。
【０１３７】
ｃ．光触媒処理層側遮光部
本発明に用いられる光触媒処理層側基板には、パターン状に形成された光触媒処理層側遮光部が形成されたものを用いても良い。このように光触媒処理層側遮光部を有する光触媒処理層側基板を用いることにより、エネルギー照射に際して、フォトマスクを用いたり、レーザ光による描画照射を行う必要がない。従って、光触媒処理層側基板とフォトマスクとの位置合わせが不要であることから、簡便な工程とすることが可能であり、また描画照射に必要な高価な装置も不必要であることから、コスト的に有利となるという利点を有する。
【０１３８】
このような光触媒処理層側遮光部を有する光触媒処理層側基板は、光触媒処理層側遮光部の形成位置により、下記の二つの実施態様とすることができる。
【０１３９】
一つが、例えば図９に示すように、基板１上に光触媒処理層側遮光部３１を形成し、この光触媒処理層側遮光部３１上に光触媒処理層２を形成して、光触媒処理層側基板３とする実施態様である。もう一つは、例えば図１０に示すように、基板１上に光触媒処理層２を形成し、その上に光触媒処理層側遮光部３１を形成して光触媒処理層側基板３とする実施態様である。
【０１４０】
いずれの実施態様においても、フォトマスクを用いる場合と比較すると、光触媒処理層側遮光部が、上記光触媒処理層と特性変化層とが間隙をもって位置する部分の近傍に配置されることになるので、基板内等におけるエネルギーの散乱の影響を少なくすることができることから、エネルギーのパターン照射を極めて正確に行うことが可能となる。
【０１４１】
さらに、上記光触媒処理層上に光触媒処理層側遮光部を形成する実施態様においては、光触媒処理層と特性変化層とを所定の間隙をおいて配置する際に、この光触媒処理層側遮光部の膜厚をこの間隙の幅と一致させておくことにより、上記光触媒処理層側遮光部を上記間隙を一定のものとするためのスペーサとしても用いることができるという利点を有する。
【０１４２】
すなわち、所定の間隙をおいて上記光触媒処理層と特性変化層とを接触させた状態で配置する際に、上記光触媒処理層側遮光部と特性変化層とを密着させた状態で配置することにより、上記所定の間隙を正確とすることが可能となり、そしてこの状態で光触媒処理層側基板からエネルギーを照射することにより、特性変化層上にパターンを精度良く形成することが可能となるのである。
【０１４３】
このような光触媒処理層側遮光部の形成方法は、特に限定されるものではなく、光触媒処理層側遮光部の形成面の特性や、必要とするエネルギーに対する遮蔽性等に応じて適宜選択されて用いられる。
【０１４４】
例えば、スパッタリング法、真空蒸着法等により厚み１０００〜２０００Å程度のクロム等の金属薄膜を形成し、この薄膜をパターニングすることにより形成されてもよい。このパターニングの方法としては、スパッタ等の通常のパターニング方法を用いることができる。
【０１４５】
また、樹脂バインダ中にカーボン微粒子、金属酸化物、無機顔料、有機顔料等の遮光性粒子を含有させた層をパターン状に形成する方法であってもよい。用いられる樹脂バインダとしては、ポリイミド樹脂、アクリル樹脂、エポキシ樹脂、ポリアクリルアミド、ポリビニルアルコール、ゼラチン、カゼイン、セルロース等の樹脂を１種または２種以上混合したものや、感光性樹脂、さらにはＯ／Ｗエマルジョン型の樹脂組成物、例えば、反応性シリコーンをエマルジョン化したもの等を用いることができる。このような樹脂製光触媒処理層側遮光部の厚みとしては、０．５〜１０μｍの範囲内で設定することができる。このよう樹脂製光触媒処理層側遮光部のパターニングの方法は、フォトリソ法、印刷法等一般的に用いられている方法を用いることができる。
【０１４６】
なお、上記説明においては、光触媒処理層側遮光部の形成位置として、基板と光触媒処理層との間、および光触媒処理層表面の二つの場合について説明したが、その他、基板の光触媒処理層が形成されていない側の表面に光触媒処理層側遮光部を形成する態様も採ることが可能である。この態様においては、例えばフォトマスクをこの表面に着脱可能な程度に密着させる場合等が考えられる。
【０１４７】
ｄ．プライマー層
本発明において、上述したように基板上に光触媒処理層側遮光部をパターン状に形成して、その上に光触媒処理層を形成して光触媒処理層側基板とする場合においては、上記光触媒処理層側遮光部と光触媒処理層との間にプライマー層を形成することが好ましい。
【０１４８】
このプライマー層の作用・機能は必ずしも明確なものではないが、光触媒処理層側遮光部と光触媒処理層との間にプライマー層を形成することにより、プライマー層は光触媒の作用による特性変化層の特性変化を阻害する要因となる光触媒処理層側遮光部および光触媒処理層側遮光部間に存在する開口部からの不純物、特に、光触媒処理層側遮光部をパターニングする際に生じる残渣や、金属、金属イオン等の不純物の拡散を防止する機能を示すものと考えられる。従って、プライマー層を形成することにより、高感度で特性変化の処理が進行し、その結果、高解像度のパターンを得ることが可能となるのである。
【０１４９】
なお、本発明においてプライマー層は、光触媒処理層側遮光部のみならず光触媒処理層側遮光部間に形成された開口部に存在する不純物が光触媒の作用に影響することを防止するものであるので、プライマー層は開口部を含めた光触媒処理層側遮光部全面にわたって形成されていることが好ましい。
【０１５０】
図１１はこのようなプライマー層を形成した光触媒処理層側基板の一例を示すものである。光触媒処理層側遮光部３１が形成された基板１の光触媒処理層側遮光部３１が形成されている側の表面にプライマー層３２が形成されており、このプライマー層３２の表面に光触媒処理層２が形成されている。
【０１５１】
上記基板上に光触媒処理層側遮光部がパターン状に形成された構成は、一般的なフォトマスクの構成である。従って、このプライマー層は、光触媒処理層がプライマー層を介してフォトマスク上に形成されたものであるといえる。
【０１５２】
本発明におけるプライマー層は、光触媒処理層とフォトマスクとが物理的に接触しないように配置された構造であれば特に限定されるものではない。すなわち、フォトマスクの光触媒処理層側遮光部と光触媒処理層とが接触しないようにプライマー層が形成されていればよいのである。
【０１５３】
このプライマー層を構成する材料としては、特に限定されるものではないが、光触媒の作用により分解されにくい無機材料が好ましい。具体的には無定形シリカを挙げることができる。このような無定形シリカを用いる場合には、この無定形シリカの前駆体は、一般式ＳｉＸ_４で示され、Ｘはハロゲン、メトキシ基、エトキシ基、またはアセチル基等であるケイ素化合物であり、それらの加水分解物であるシラノール、または平均分子量３０００以下のポリシロキサンが好ましい。
【０１５４】
また、プライマー層の膜厚は、０．００１μｍから１μｍの範囲内であることが好ましく、特に０．００１μｍから０．１μｍの範囲内であることが好ましい。
【０１５５】
（２）エネルギー照射
本発明においては、光触媒処理層側基板とマイクロアレイチップ用基材とを、光触媒処理層および特性変化層が向かい合うように、かつ２００μｍ以下の間隙をおいて配置した後、所定の方向からエネルギーを照射するパターン形成工程が行われることを特徴とする。なお、２００μｍ以下の間隙とは、光触媒処理層および特性変化層が接触している状態も含むものとする。
【０１５６】
このように光触媒処理層と特性変化層表面を所定の間隔で離して配置することにより、酸素と水および光触媒作用により生じた活性酸素種が脱着しやすくなる。すなわち、上記範囲より間隔を離して配置した場合は、生じた活性酸素種が特性変化層に届き難くなり、特性の変化速度を遅くしてしまう可能性があることから好ましくないのである。逆に、光触媒処理層および特性変化層との間隔を狭くしすぎると、酸素と水および光触媒作用により生じた活性酸素種の脱着がしにくくなり、結果的に特性の変化速度を遅くしてしまう可能性があることから好ましくない。
【０１５７】
本発明において上記間隙は、パターン精度が極めて良好であり、光触媒の感度も高く、従って特性変化の効率が良好である点を考慮すると特に０．２μｍ〜１０μｍの範囲内、好ましくは１μｍ〜５μｍの範囲内とすることが好ましい。
【０１５８】
一方、例えば３００ｍｍ×３００ｍｍといった大面積のマイクロアレイチップ用基材に対して処理を行う場合は、接触することなく、かつ上述したような微細な間隙を光触媒処理層側基板とマイクロアレイチップ用基材との間に設けることは極めて困難である。したがって、マイクロアレイチップ用基材が比較的大面積である場合は、上記間隙は、１０〜１００μｍの範囲内、特に５０〜７５μｍの範囲内とすることが好ましい。間隙をこのような範囲内とすることにより、パターンがぼやける等のパターン精度の低下の問題や、光触媒の感度が悪化して特性変化の効率が悪化する等の問題が生じることなく、さらに特性変化層上の特性変化のムラが発生しないといった効果を有するからである。
【０１５９】
このように比較的大面積のマイクロアレイチップ用基材にエネルギー照射する際には、エネルギー照射装置内の光触媒処理層側基板とマイクロアレイチップ用基材との位置決め装置における間隙の設定を、１０μｍ〜２００μｍの範囲内、特に２５μｍ〜７５μｍの範囲内に設定することが好ましい。設定値をこのような範囲内とすることにより、パターン精度の大幅な低下や光触媒の感度の大幅な悪化を招くことなく、かつ光触媒処理層側基板とマイクロアレイチップ用基材とが接触することなく配置することが可能となるからである。
【０１６０】
本発明においては、このような間隙をおいた配置状態は、少なくともエネルギー照射の間だけ維持されればよい。
【０１６１】
このような極めて狭い間隙を均一に形成して光触媒処理層と特性変化層とを配置する方法としては、例えばスペーサを用いる方法を挙げることができる。そして、このようにスペーサを用いることにより、均一な間隙を形成することができると共に、このスペーサが接触する部分は、光触媒の作用が特性変化層表面に及ばないことから、このスペーサを上述したパターンと同様のパターンを有するものとすることにより、特性変化層上に所定のパターンを形成することが可能となる。
【０１６２】
本発明においては、このようなスペーサを一つの部材として形成してもよいが、工程の簡略化等のため、上記光触媒処理層側基板の欄で説明したように、光触媒処理層側基板の光触媒処理層表面に形成することが好ましい。なお、上記光触媒処理層側基板における説明においては、光触媒処理層側遮光部として説明したが、本発明においては、このようなスペーサは特性変化層表面に光触媒の作用が及ばないように表面を保護する作用を有すればよいものであることから、特に照射されるエネルギーを遮蔽する機能を有さない材料で形成されたものであってもよい。
【０１６３】
次に、上述したような接触状態を維持した状態で、接触する部分へのエネルギー照射が行われる。通常このようなエネルギー照射に用いる光の波長は、４００ｎｍ以下の範囲、好ましくは３８０ｎｍ以下の範囲から設定される。これは、上述したように光触媒処理層に用いられる好ましい光触媒が二酸化チタンであり、この二酸化チタンにより光触媒作用を活性化させるエネルギーとして、上述した波長の光が好ましいからである。
【０１６４】
このようなエネルギー照射に用いることができる光源としては、水銀ランプ、メタルハライドランプ、キセノンランプ、エキシマランプ、その他種々の光源を挙げることができる。
【０１６５】
上述したような光源を用い、フォトマスクを介する等のパターン照射により行う方法の他、エキシマ、ＹＡＧ等のレーザを用いてパターン状に描画照射する方法を用いることも可能である。
【０１６６】
また、エネルギー照射に際してのエネルギーの照射量は、特性変化層表面が光触媒処理層中の光触媒の作用により特性変化層表面の特性の変化が行われるのに必要な照射量とする。
【０１６７】
この際、光触媒処理層を加熱しながらエネルギー照射することにより、感度を上昇させことが可能となり、効率的な特性の変化を行うことができる点で好ましい。具体的には３０℃〜８０℃の範囲内で加熱することが好ましい。
【０１６８】
本発明におけるエネルギーの照射方向は、光触媒処理層側基板に光触媒処理層側遮光部が形成されているか否か、光触媒処理層側基板もしくはマイクロアレイチップ用基材が透明であるか否かにより決定される。
【０１６９】
すなわち、光触媒処理層側基板に光触媒処理層側遮光部が形成されている場合は、光触媒処理層側基板側からエネルギーを照射する必要があり、かつこの場合は光触媒処理層側基板が照射されるエネルギーに対して透明である必要がある。なお、この場合、光触媒処理層上に光触媒処理層側遮光部が形成され、かつこの光触媒処理層側遮光部を上述したようなスペーサとしての機能を有するように用いた場合においては、照射方向は光触媒処理層側基板側からでもマイクロアレイチップ用基材側からであってもよい。
【０１７０】
また、光触媒処理層がパターン状に形成されている場合における照射方向は、上述したように、光触媒処理層と特性変化層とが接触する部分にエネルギーが照射されるのであればいかなる方向から照射されてもよい。
【０１７１】
同様に、上述したスペーサを用いる場合も、接触する部分にエネルギーが照射されるのであればいかなる方向から照射されてもよい。
【０１７２】
フォトマスクを用いる場合は、フォトマスクが配置された側からエネルギーが照射される。この場合は、フォトマスクが配置された側の基板または基材、すなわち光触媒処理層側基板もしくはマイクロアレイチップ用基材のいずれかが透明である必要がある。
【０１７３】
上述したようなエネルギー照射が終了すると、光触媒処理層側基板が特性変化層との接触位置から離され、これにより例えば、図１（ｃ）に示すように濡れ性が変化した親水性領域８および撥水性領域９からなるパターンが濡れ性変化層５上に形成される。一方、後述する第２実施態様における分解除去層の場合は、図２（ｃ）に示すように、パターン状に形成された分解除去層２１を得る。
【０１７４】
Ｃ．取り外し工程
本態様においては、エネルギー照射が行われた後、マイクロアレイチップ用基材から光触媒処理層基板が取り外す。これにより表面に親水性領域および撥水性領域からなるパターンが形成された濡れ性変化層を得る。
【０１７５】
Ｄ．固定化層形成工程
次に、本態様における固定化層形成工程について説明する。本工程は、親水性領域および撥水性領域からなる濡れ性変化層のパターンに従い、親水性領域上に固定化層を形成する工程である。
【０１７６】
以下、本工程によりパターン状に形成される固定化層について説明する。
【０１７７】
（固定化層）
本態様においては、上述した濡れ性変化層上にパターン状に形成された親水性領域上に、後述する特異性生体高分子を固定化するための固定化層が形成される。
【０１７８】
この固定化層に用いられる固定化剤としては、特異性生体高分子の固定化手段に応じて種々のものを用いることができる。
【０１７９】
このような固定化手段としては、当業者において周知の結合方法を使用することができ、例えば、静電結合による固定化方法、共有結合による固定化方法等を使用することができる。
【０１８０】
上記静電結合による固定化手段の場合は、特異性生体高分子が有する電荷と反対の電荷を有する物質を固定化剤として溶解した溶液を固定化層形成用塗工液として調製し、これを上記親水性領域がパターン状に形成された濡れ性変化層上に塗布することにより固定化層を形成する。
【０１８１】
例えば、特異性生体高分子がオリゴヌクレオチドの場合は、オリゴヌクレオチドが陰イオン（アニオン）性を有するものであるので、陽イオン（カチオン）性を有する物質を固定化剤とした塗工液が用いられ、これを塗布して乾燥することにより固定化層が形成される。
【０１８２】
この際、用いられる陽イオン性を有する固定化剤としては、ポリＬ‐リシン、ポリ（アリルアミンヒドロクロリド）、ポリ（エチレンイミン）、ポリ（ジアリルジメチルアンモニウムクロリド）等を挙げることができ、中でもポリＬ‐リシンが好ましい。
【０１８３】
一方、共有結合（化学結合）による固定化法としては、官能基（例えば、カルボキシル基、アミノ基、又は水酸基）を有する化合物を固定化剤として固定化剤形成用塗料に用いて固定化層を形成し、これに特異性生体高分子を結合させる方法を挙げることができる。このような固定化剤としては、官能基としてカルボキシル基又は第一級アミノ基をもつ化合物を挙げることができる。
【０１８４】
具体的には、アミノ基を含むシランカップリング剤を、濡れ性変化層上にパターン状に形成された親水性領域に付着させる。これは、上記シランカップリング剤を含む溶液を用い、ディップコーティング法、転写法、スピンコーティング法等により全面に塗布し、親水性領域のみにシランカップリング剤を付着させる方法、インクジェット、ディスペンサ等を用いて親水性領域のみを狙って塗布する方法を挙げることができる。コーティングされたアミノ基を含むシランカップリング剤は、親水性領域に濡れ広がり、撥水性領域に残留することはない。コーティング後、必要に応じて加熱することにより、親水性領域と固定化剤とを結合させることも可能である。そして、このようにして形成した固定化層上に特異性生体高分子を結合させることにより、マイクロアレイチップを形成することができる。
【０１８５】
上記固定化剤であるシランカップリング剤のアミノ基と、末端をアミノ基で修飾した生体高分子、具体的にはオリゴヌクレオチド等の末端アミノ基とを共有結合させることにより強固に膜に固定化することができる。
【０１８６】
この際、用いられるアミノ基を有するシランカップリング剤としては、Ｎ−β（アミノエチル）γ−アミノプロピルトリメトキシシラン、Ｎ−β（アミノエチル）γ−アミノプロピルメチルジメトキシシラン、γ−アミノプロピルトリエトキシシラン、Ｎ−フェニル−γ−アミノプロピルトリメトキシシラン等を挙げることができる。
【０１８７】
また、固定化層と特異性生体高分子との共有結合による別の固定化法としては、例えば、生体高分子がオリゴヌクレオチドの場合、その５'末端を上記官能基に共有結合で結合させる方法が知られている。具体的には、まず、オリゴヌクレオチドの５'末端とマレイミド化合物とを反応させ、オリゴヌクレオチドの５'末端にマレイミド基を導入する。好適なマレイミド化合物としては、スルホスクシンイミジル−４−（Ｎ−マレイミドメチル）シクロヘキサン−１−カルボキシレート等を挙げることができる。上記反応とは別に、アミノ基を官能基として有する固定化剤とスクシンイミジル−Ｓ−アセチルチオアセテートとを反応させた後に、ヒドロキシラミンを用いて脱アセチル化を行なうことにより、上記固定化剤にＳＨ基を付与する。こうして付与した固定化剤上のＳＨ基と、５'末端にマレイミド基を導入して調製した上記オリゴヌクレオチドの５'末端に導入したマレイミド基とを反応させることにより、オリゴヌクレオチドと固定化剤とを結合させることができ、特異性生体高分子を固定化層に固定することができるのである。
【０１８８】
その他、上述した方法の他にも、多くの固定化方法、例えば、ビオチン−アビジン系を用いる方法も使用することができる。
【０１８９】
本態様に用いられる固定化層は、後述するように、その上に特異性生体高分子を含む液体を付着させた際に、この液体が固定化層より広がらないことが好ましい。従って、固定化層の表面の濡れ性が、周囲の濡れ性変化層の領域（撥水性領域）より、水との接触角で１度以上、好ましくは５度以上小さくなるような固定化剤が選択されて用いられることが好ましい。
【０１９０】
Ｅ．特異性生体高分子固定工程
最後に、本態様における特異性生体高分子固定工程について説明する。本工程は、上記固定化層形成工程によりパターン状に形成された固定化層上に、特異性生体高分子を付着させる工程である。
【０１９１】
以下、本工程において形成される特異性生体高分子について説明する。
【０１９２】
（特異性生体高分子）
本態様のマイクロアレイチップは、上記固定化層上に特異性生体高分子が固定化されて配置される。このような特異性生体高分子としては、所定の生体高分子と特異的に結合することができる生体高分子であれば特に限定されるものではない。
【０１９３】
このような特異的に結合する場合としては、例えば、核酸（例えば、オリゴヌクレオチド又はポリヌクレオチド）とそれに相補的な核酸との組み合わせ、抗原と抗体（又は抗体フラグメント）との組み合わせ、受容体とそのリガンド（例えば、ホルモン、サイトカイン、神経伝達物質、又はレクチン）との組み合わせ、酵素とそのリガンド（例えば、酵素の基質アナログ、補酵素、調節因子、又は阻害剤）との組み合わせ、酵素アナログとその酵素アナログの元となる酵素の基質との組み合わせ、又はレクチンと糖との組み合わせ等を挙げることができる。なお、「酵素アナログ」とは、元の酵素に対する基質との特異的な親和性は高いものの、触媒活性は示さないものを意味する。また、上記の各組み合わせにおける各化合物は、それぞれ、いずれか一方が「特異性生体高分子」となり、他方が、検出される物質となることができる。例えば、「抗原と抗体との組み合わせ」では、抗原が「検出される物質」となる場合には、抗体が「特異性生体高分子」となることができ、逆に、抗体が「検出される物質」となる場合には、抗原が「特異性生体高分子」となることができる。
【０１９４】
例えば、本態様のマイクロアレイチップを、核酸ハイブリダイゼーションアッセイに適用する場合には、上記特異性生体高分子として、検出される物質である核酸（例えば、オリゴヌクレオチド又はポリヌクレオチド）と相補的に結合することのできるオリゴヌクレオチド又はポリヌクレオチドを用いることができる。本明細書においては、「オリゴヌクレオチド」又は「ポリヌクレオチド」には、２'−デオキシリボ核酸（ＤＮＡ）、リボ核酸（ＲＮＡ）、及びペプチド核酸（ＰＮＡ）が含まれる。なお、ＰＮＡとは、ＤＮＡのホスホジエステル結合をペプチド結合に変換した人工核酸である。上記不溶性粒子に結合されるオリゴヌクレオチド又はポリヌクレオチドの鎖長は、分析目的に応じて適宜選択することができ、例えば、捕捉しようとするＤＮＡ、ＲＮＡ、又はＰＮＡの相補的配列の鎖長に基づいて決定することができる。
【０１９５】
上記特異性生体高分子として用いるオリゴヌクレオチドの合成は、自動合成装置を用いて一般的に行なうことができる。また、必要に応じて、自動合成装置を用いて、末端アミノ基又は他の基が修飾されたオリゴヌクレオチドを製造することもできる。あるいは、自動合成装置を用いて未修飾オリゴヌクレオチドを予め合成しておき、必要に応じて、上記の未修飾オリゴヌクレオチドを修飾することもできる。オリゴヌクレオチドを修飾する方法は周知であり、例えば、アミン類やフルオレセイン等を付加することができる。このように修飾されたオリゴヌクレオチドは、未修飾のオリゴヌクレオチドと比べて、固定化層に対する親和性が大きい。また、特異性生体高分子として用いるポリヌクレオチド（例えば、ｃＤＮＡ）の合成は、通常の遺伝子工学的手法、例えば、ＰＣＲ（ポリメラーゼ連鎖反応）法を用いて行なうことができる。
【０１９６】
また、本態様のマイクロアレイチップを免疫学的アッセイに適用する場合には、上記特異性生体高分子として、検出される物質と特異的に結合する抗原（ハプテンを含む）又は抗体を用いることができる。この場合に、上記検出される物質としては、被検試料中に一般的に含まれている成分で、しかも、免疫学的に検出することのできる物質あれば、特に制限されない。一例を挙げれば、各種タンパク質、多糖類、脂質、菌体、又は各種環境物質等を挙げることができる。より詳細には、免疫グロブリン（例えば、ＩｇＧ、ＩｇＭ、又はＩｇＡ）、感染症関連マーカー（例えば、ＨＢｓ抗原、ＨＢｓ抗体、ＨＩＶ−１抗体、ＨＩＶ−２抗体、ＨＴＬＶ−１抗体、又はトレポネーマ抗体）、腫瘍関連抗原（例えば、ＡＦＰ、ＣＲＰ、又はＣＥＡ）、凝固線溶マーカー（例えば、プラスミノーゲン、アンチトロンビン−III、Ｄ−ダイマー、ＴＡＴ、又はＰＰＩ）、抗てんかん薬（例えば、ホルモン）、各種薬剤（例えば、ジゴキシン）、菌体（例えば、Ｏ−１５７又はサルモネラ）若しくはそれらの菌体内毒素若しくは菌体外毒素、微生物類、酵素類、残留農薬、又は環境ホルモン等を挙げることができる。上記特異性生体高分子として用いる抗体としては、周知の方法で得られるポリクローナル抗体又はモノクローナル抗体のいずれをも使用することができる。さらに、上記抗体は、タンパク質［例えば、酵素（例えば、ペプシン又はパパイン）］処理したもの［例えば、抗体フラグメント（例えば、Ｆａｂ、Ｆａｂ'、Ｆ（ａｂ'）₂、又はＦｖ）］を用いることもできる。
【０１９７】
２．第２実施態様
次に本発明の第２実施態様におけるマイクロアレイチップの製造方法について説明する。
【０１９８】
第２実施態様は、エネルギー照射に伴う光触媒の作用により分解除去する分解除去層を用いて、固定化層のパターニングを行う態様である。さらに、本態様においては、分解除去層と基材との濡れ性の差を利用して固定化層を形成する態様と、分解除去層自体を固定化層として用いる態様とに分けることができる。以下、各々の態様に分けて説明する。
【０１９９】
Ａ．分解除去層と基材との濡れ性の差を利用して固定化層を形成する場合
このようなマイクロアレイチップの製造方法は、基材上に、エネルギー照射に伴う光触媒の作用により分解除去され、かつ前記基材とは液体との接触角が異なる分解除去層を形成し、マイクロアレイチップ用基材を調整するマイクロアレイチップ用基材調整工程と、基板上に少なくとも光触媒を含有する光触媒処理層が形成されている光触媒処理層側基板を用い、前記マイクロアレイチップ用基材および前記光触媒処理層側基板を、前記光触媒処理層および前記分解除去層が向かい合うように、かつ、２００μｍ以下の間隙をおいて配置した後、所定の方向からエネルギーを照射することにより、前記分解除去層をパターン状に形成するパターン形成工程と、前記マイクロアレイチップ用基材から前記光触媒処理層側基板を取り外す取り外し工程と、前記分解除去層のパターンに従い、前記分解除去層上または前記基材上に固定化層を形成する固定化層形成工程と、前記固定化層上に特異性生体高分子を固定させる特異性生体高分子固定工程とを少なくとも有することを特徴とするものである。
【０２００】
このような本態様におけるマイクロアレイチップの製造方法について図面を用いて説明する。
【０２０１】
図２は、本態様のマイクロアレイチップの製造方法の一例を示している。まず、少なくとも光触媒からなる光触媒処理層２が基板１上に成膜されている光触媒処理層側基板３を準備する。さらに、当該光触媒処理層側基板３とは別に、基材４を準備し、当該基材４上にエネルギー照射に伴う光触媒の作用により分解除去され、かつ基材４とは異なる濡れ性を有する分解除去層２１を成膜し、マイクロアレイチップ用基材６を準備する（図２（ａ）参照）。
【０２０２】
さらに、光触媒処理層側基板３およびマイクロアレイチップ用基材６を、光触媒処理層２および分解除去層２１が向かい合うように、かつ所定の間隙を有して配置した後、フォトマスク７を介してエネルギーをパターン状に照射する（図２（ｂ）参照）。
【０２０３】
次いで、マイクロアレイチップ用基材６から光触媒処理層側基板３を取り外すと、基材４上にパターン状に形成された分解除去層２１を有するマイクロアレイチップ用基材６を得る（図２（ｃ）参照）。
【０２０４】
また、分解除去層２１と基材４とは、濡れ性が異なることから、パターン状に分解除去層２１を形成することにより、基材４上には濡れ性の違いによるパターンが形成されたことになる。この濡れ性の違いを利用し、分解除去層２１が分解除去され露出している基材４上に固定化層１０を形成する（図２（ｄ）参照）。なお、図２では、分解除去層２１が分解除去され露出している基材４上に固定化層１０を形成する場合を示しているが、このような場合に限らず、例えば、分解除去層よりも基材のほうが液体に対する接触角が大きい場合には、分解除去層上に固定化層が形成される。
【０２０５】
最後に、固定化層１０上に、特異性生体高分子１１を付着させることにより、マイクロアレイチップを製造することができる（図２（ｅ）参照）。
【０２０６】
このような本態様によれば、特にエネルギー照射後の後処理が必要無く、さらに、エネルギー照射後、マイクロアレイチップから光触媒処理層側基板を取り外すので、マイクロアレイチップ自体には光触媒処理層が含まれることがない。従って、マイクロアレイチップに光触媒の半永久的な作用が及ぶことを皆無とするいことができるのである。
【０２０７】
以下、このような場合の本態様のマイクロアレイチップの製造方法について各工程ごとに詳細に説明する。
【０２０８】
（１）マイクロアレイチップ用基材調整工程
本態様におけるマイクロアレイチップ用基材調整工程とは、基材上に、エネルギー照射に伴う光触媒の作用により分解除去され、かつ前記基材とは液体との接触角が異なる分解除去層を形成し、マイクロアレイチップ用基材を調整する工程である。
【０２０９】
このような本工程において形成されるマイクロアレイチップ用基材について説明する。
【０２１０】
（マイクロアレイチップ用基材）
ａ．分解除去層
まず、マイクロアレイチップ用基材を構成する分解除去層について説明する。本態様における分解除去層は、エネルギー照射の際に光触媒処理層中の光触媒の作用により、エネルギー照射された部分の分解除去層が分解除去される層であり、かつ、分解除去層が分解除去された際に露出する基材と異なる濡れ性を示す層である。
【０２１１】
このような分解除去層は、エネルギー照射した部分が光触媒の作用により分解除去されることから、現像工程や洗浄工程を行うことなく分解除去層のある部分と無い部分からなるパターン、すなわち凹凸を有するパターンを形成することができる。
【０２１２】
なお、この分解除去層は、エネルギー照射による光触媒の作用により酸化分解され、気化等されることから、現像・洗浄工程等の特別な後処理なしに除去されるものであるが、分解除去層の材質によっては、洗浄工程等を行ってもよい。
【０２１３】
また本態様は、パターン状に形成された分解除去層を用い、かつ分解除去層と基材との濡れ性の差を利用して、固定化層をパターニングする態様であることから、分解除去層と基材との液体に対する接触角が異なるように構成されている。
【０２１４】
このような分解除去層が有する濡れ性としては、固定化層を分解除去層が分解除去され露出している基材上に形成する場合には、基材の液体に対する接触角よりも、分解除去層の液体に対する接触角を大きくする。
【０２１５】
このような場合、塗布される固定化層形成用塗工液が有する表面張力と同等の表面張力の液体に対する分解除去層の接触角が、基材のそれよりも、１°以上、特に５°以上、中でも１０°以上大きいことが好ましい。
【０２１６】
このような場合、分解除去層表面に要求される撥水性としては、塗布される固定化層形成用塗工液が有する表面張力と同等の表面張力の液体に対する接触角が、３０°以上、特に４０°以上、中でも５０°以上となるものであることが好ましい。
【０２１７】
このように、分解除去層が上記範囲内の撥水性を有し、基材と分解除去層が上記範囲内の濡れ性の差を有していれば、固定化層形成用塗工液を塗布した際に、分解除去層上に付着することがなく、基材上に固定化層を精度良くパターン状に形成することができるからである。
【０２１８】
一方、固定化層を分解除去層上に形成する場合には、基材の液体に対する接触角よりも、分解除去層の液体に対する接触角を小さくする。これにより、固定化層を形成する固定化層形成用塗工液が、分解除去層が分解除去され露出している基材上に付着することが少なく、分解除去層上に良好に付着させることができるからである。
【０２１９】
なお、ここでいう液体との接触角は、種々の表面張力を有する液体との接触角を接触角測定器（協和界面科学（株）製ＣＡ−Ｚ型）を用いて測定（マイクロシリンジから液滴を滴下して３０秒後）し、その結果から、もしくはその結果をグラフにして得たものである。また、この測定に際して、種々の表面張力を有する液体としては、純正化学株式会社製のぬれ指数標準液を用いた。
【０２２０】
また、本態様における分解除去層の膜厚としては、後述するパターン形成工程において、照射されるエネルギーに伴う光触媒の作用により分解除去することが可能な膜厚であれば特に限定されない。具体的には、０．００１μｍ〜１μｍであることが好ましく、特に好ましくは０．０１〜０．１μｍの範囲内である。
【０２２１】
このような分解除去層に用いることができる材料としては、上述した分解除去層の特性、すなわちエネルギー照射に伴う光触媒の作用により分解除去される材料であり、かつ好ましくは、上記範囲内の液体との接触角を有する材料である。
【０２２２】
上記分解除去層の成膜方法としては、特に限定されるものではないが、上述したような材料を選択し、適切な溶剤に希釈させ塗工液としたものをスピンコーティング等の一般的な塗布法を用いて成膜する方法を挙げることができる。このような薄膜の形成方法は、容易でかつコスト的に有利な方法であるといえる。一方、機能性薄膜、すなわち、自己組織化単分子膜、ラングミュア−ブロケット膜、および交互吸着膜等の成膜方法により形成することも可能である。これらの方法は、緻密で欠陥の少ない薄膜の形成を可能とするため好ましい方法であるといえる。
【０２２３】
ここで、本態様に用いられる自己組織化単分子膜、ラングミュア−ブロケット膜、および交互吸着膜について具体的に説明する。
【０２２４】
▲１▼ 自己組織化単分子膜
自己組織化単分子膜(Self-Assembled Monolayer)の公式な定義の存在を発明者らは知らないが、一般的に自己組織化膜として認識されているものの解説文としては、例えばAbraham Ulmanによる総説"Formation and Structure of Self-Assembled Monolayers", Chemical Review, 96, 1533-1554 (1996)が優れている。本総説を参考にすれば、自己組織化単分子膜とは、適当な分子が適当な基材表面に吸着・結合（自己組織化）した結果生じた単分子層のことと言える。自己組織化膜形成能のある材料としては、例えば、脂肪酸などの界面活性剤分子、アルキルトリクロロシラン類やアルキルアルコキシド類などの有機ケイ素分子、アルカンチオール類などの有機イオウ分子、アルキルフォスフェート類などの有機リン酸分子などが挙げられる。分子構造の一般的な共通性は、比較的長いアルキル鎖を有し、片方の分子末端に基材表面と相互作用する官能基が存在することである。アルキル鎖の部分は分子同士が２次元的にパッキングする際の分子間力の源である。もっとも、ここに示した例は最も単純な構造であり、分子のもう一方の末端にアミノ基やカルボキシル基などの官能基を有するもの、アルキレン鎖の部分がオキシエチレン鎖のもの、フルオロカーボン鎖のもの、これらが複合したタイプの鎖のものなど様々な分子から成る自己組織化単分子膜が報告されている。また、複数の分子種から成る複合タイプの自己組織化単分子膜もある。また、最近では、デンドリマーに代表されるような粒子状で複数の官能基（官能基が一つの場合もある）を有する高分子や直鎖状（分岐構造のある場合もある）の高分子が一層基材表面に形成されたもの（後者はポリマーブラシと総称される）も自己組織化単分子膜と考えられる場合もあるようである。本発明は、これらも自己組織化単分子膜に含める。
【０２２５】
▲２▼ ラングミュア−ブロジェット膜
本態様に用いられるおけるラングミュア−ブロジェット膜(Langmuir-Blodgett Film)は、基材上に形成されてしまえば形態上は上述した自己組織化単分子膜との大きな相違はない。ラングミュア−ブロジェット膜の特徴はその形成方法とそれに起因する高度な２次元分子パッキング性（高配向性、高秩序性）にあると言える。すなわち、一般にラングミュア−ブロジェット膜形成分子は気液界面上に先ず展開され、その展開膜がトラフによって凝縮されて高度にパッキングした凝縮膜に変化する。実際は、これを適当な基材に移しとって用いる。ここに概略を示した手法により単分子膜から任意の分子層の多層膜まで形成することが可能である。また、低分子のみならず、高分子、コロイド粒子なども膜材料とすることができる。様々な材料を適用した最近の事例に関しては宮下徳治らの総説“ソフト系ナノデバイス創製のナノテクノロジーへの展望” 高分子 50巻 9月号 644-647 (2001)に詳しく述べられている。
【０２２６】
▲３▼ 交互吸着膜
交互吸着膜(Layer-by-Layer Self-Assembled Film)は、一般的には、最低２個の正または負の電荷を有する官能基を有する材料を逐次的に基材上に吸着・結合させて積層することにより形成される膜である。多数の官能基を有する材料の方が膜の強度や耐久性が増すなど利点が多いので、最近ではイオン性高分子（高分子電解質）を材料として用いることが多い。また、タンパク質や金属や酸化物などの表面電荷を有する粒子、いわゆる“コロイド粒子”も膜形成物質として多用される。さらに最近では、水素結合、配位結合、疎水性相互作用などのイオン結合よりも弱い相互作用を積極的に利用した膜も報告されている。比較的最近の交互吸着膜の事例については、静電的相互作用を駆動力にした材料系に少々偏っているがPaula T. Hammondによる総説"Recent Explorations in Electrostatic Multilayer Thin Film Assembly" Current Opinion in Colloid & Interface Science, 4, 430-442 (2000)に詳しい。交互吸着膜は、最も単純なプロセスを例として説明すれば、正（負）電荷を有する材料の吸着−洗浄−負（正）電荷を有する材料の吸着−洗浄のサイクルを所定の回数繰り返すことにより形成される膜である。ラングミュア−ブロジェット膜のように展開−凝縮−移し取りの操作は全く必要ない。また、これら製法の違いより明らかなように、交互吸着膜はラングミュア−ブロジェット膜のような２次元的な高配向性・高秩序性は一般に有さない。しかし、交互吸着膜及びその作製法は、欠陥のない緻密な膜を容易に形成できること、微細な凹凸面やチューブ内面や球面などにも均一に成膜できることなど、従来の成膜法にない利点を数多く有している。
【０２２７】
ｂ．基材
次に、本態様の基材について説明する。本態様においては、上述した分解除去層と基材との濡れ性の差を利用して、固定化層をパターン状に形成することから、基材上に固定化層を形成する場合には、上述した分解除去層の液体に対する接触角よりも、小さい接触角を呈する基材を用いる。一方、上述した分解除去層上に固定化層を形成する場合には、上述した分解除去層の呈する液体に対する接触角よりも、大きい接触角を呈する基材を用いる。
【０２２８】
（２）パターン形成工程
本態様におけるパターン形成工程とは、基板上に少なくとも光触媒を含有する光触媒処理層が形成されている光触媒処理層側基板を用い、前記マイクロアレイチップ用基材および前記光触媒処理層側基板を、前記光触媒処理層および前記分解除去層が向かい合うように、かつ、２００μｍ以下の間隙をおいて配置した後、所定の方向からエネルギーを照射することにより、前記分解除去層をパターン状に形成する工程である。
【０２２９】
このような本工程については、上述した第１実施態様と同様であるので、ここでの説明は省略する。
【０２３０】
（３）取り外し工程
本態様においては、エネルギー照射が行われた後、マイクロアレイチップ用基材から光触媒処理層基板を取り外す。これにより、基材上にパターン状に形成された分解除去層を有する基材を得る。
【０２３１】
（４）固定化層形成工程
次に固定化層形成工程を行う。本態様における固定化層形成工程とは、パターン状に形成された分解除去層を有する基材上に、固定化層形成用塗工液を塗布し、分解除去層と基材との濡れ性の差を利用して、分解除去層上、または、分解除去層が分解除去され露出している基材上に固定化層をパターン状に形成する工程である。
【０２３２】
本態様においては、上述したように分解除去層の液体に対する接触角が基材のそれよりも小さい場合には、分解除去層上に固定化層が形成される。逆に、分解除去層の液体に対する接触角が基材のそれよりも大きい場合には、上述したパターン形成工程において分解除去層が分解除去され露出している基材上に固定化層が形成される。
【０２３３】
その他、本工程で形成される固定化層に関することは、上述した第１実施態様における「Ｄ．固定化層形成工程」の中に記載したことと同様なので、ここでの説明は省略する。
【０２３４】
（５）特異性生体高分子固定工程
本態様における特異性生体高分子固定工程とは、上述した固定化層形成工程により形成された固定化層上に、特異性生体高分子を固定させる工程である。
【０２３５】
このような本工程においても、上述した第１実施態様における「Ｅ．特異性生体高分子固定工程」と同様なのでここでの説明は省略する。
【０２３６】
Ｂ．分解除去層を固定化層として用いる場合
次に、固定化層として機能する分解除去層を用いた場合の本態様のマイクロアレイチップの製造方法について説明する。
【０２３７】
このような場合の本態様におけるマイクロアレイチップの製造方法は、基材上に、エネルギー照射に伴う光触媒の作用により分解除去され、かつ固定化層として機能する分解除去層を形成し、マイクロアレイチップ用基材を調整するマイクロアレイチップ用基材調整工程と、基板上に少なくとも光触媒を含有する光触媒処理層が形成されている光触媒処理層側基板を用い、前記マイクロアレイチップ用基材および前記光触媒処理層側基板を、前記光触媒処理層および前記分解除去層が向かい合うように、かつ、２００μｍ以下の間隙をおいて配置した後、所定の方向からエネルギーを照射することにより、固定化層としての機能を有する前記分解除去層をパターン状に形成するパターン形成工程と、前記マイクロアレイチップ用基材から前記光触媒処理層側基板を取り外す取り外し工程とを少なくとも有することを特徴とするものである。
【０２３８】
このような場合の本態様について図面を用いて説明する。
【０２３９】
図３は、固定化層としての機能を有する分解除去層を用いてマイクロアレイチップを製造する製造方法の一例を示している。まず、まず、少なくとも光触媒からなる光触媒処理層２が基板１上に成膜されている光触媒処理層側基板３を準備する。さらに、当該光触媒処理層側基板３とは別に、基材４を準備し、当該基材４上にエネルギー照射に伴う光触媒の作用により分解除去され、かつ固定化層として機能する分解除去層２１を成膜し、マイクロアレイチップ用基材６を準備する（図３（ａ）参照）。
【０２４０】
さらに、光触媒処理層側基板３およびマイクロアレイチップ用基材６を、光触媒処理層２および分解除去層２１を向かい合わせ、かつ所定の間隙を有するように配置した後、フォトマスク７を介してエネルギーをパターン状に照射する（図３（ｂ）参照）。
【０２４１】
次いで、マイクロアレイチップ用基材６から光触媒処理層側基板３を取り外すと、パターン状に形成された分解除去層２１を有する基材４を得る（図３（ｃ）参照）。本態様においては、固定化層としての機能を有する分解除去層を用いているため、このようにパターン状に形成された分解除去層を固定化層として用いることができる。
【０２４２】
そして、この固定化層としての機能を有する分解除去層２１上に、特異性生体高分子１１を付着させることにより（図３（ｄ）参照）、マイクロアレイチップを製造することができる。
【０２４３】
この場合の本態様によれば、固定化層としての機能を有する分解除去層を用いているので、分解除去層をパターン状に形成することにより固定化層をもパターン状に形成したこととなる。従って、固定化層のパターニングに要する手間が大幅に簡略化されることから、製造効率を向上させることができる。また、本態様においても、分解除去層へエネルギーを照射した後は、光触媒処理層側基板をマイクロアレイチップ用基材から取り外すため、マイクロアレイチップには光触媒処理層が含まれない。従って、本態様のマイクロアレイチップは、光触媒の作用による影響を皆無とすることができるのである。
【０２４４】
このような場合における本態様に用いる分解除去層について説明する。なお、マイクロアレイチップ用基材調整工程、パターン形成工程、取り外し工程、特異性生体高分子形成工程に関しては、上述した第２実施態様における「Ａ．分解除去層と基材との濡れ性の差を利用して固定化層を形成する場合」の中に記載したことと同様なのでここでの説明は省略する。
【０２４５】
（分解除去層）
この場合における分解除去層を形成する材料としては、上述した分解除去層の特性、すなわちエネルギー照射の際に、分解除去層と接触または所定の間隙を有して配置される光触媒処理層に含有される光触媒の作用により分解除去される材料であり、特異性生体高分子を固定させることが可能な材料であれば特に限定されない。具体的には、上述した第１実施態様における「Ｄ．固定化層形成工程」に記載した固定化層と同様の材料を用いることができる。
【０２４６】
上記分解除去層の成膜方法としては、上述したように、このような材料を、希釈し、スピンコーティング等の方法により塗布することにより薄膜を形成するといったような一般的な形成方法の他、上述した機能性薄膜、すなわち、自己組織化単分子膜、ラングミュア−ブロケット膜、および交互吸着膜等の成膜方法を用いて形成することにより、緻密で欠陥の少ない薄膜とすることができる。なお、自己組織化単分子膜、ラングミュア−ブロケット膜、および交互吸着膜については、上述したことと同様なので、ここでの説明は省略する。
【０２４７】
本態様においては、このような分解除去層を、光触媒処理層と向かい合わせ、所定の間隙を有するように配置した後、エネルギー照射することにより、パターン状に形成する。これによりパターン状の固定化層を得る。そして、この固定化層としての機能を有する分解除去層上に特異性生体高分子を付着させ、固定させることにより、マイクロアレイチップを製造することができる。
【０２４８】
ＩＩ．マイクロアレイチップ
次に、本発明のマイクロアレイチップについて説明する。
【０２４９】
本発明のマイクロアレイチップは、エネルギー照射に伴う光触媒の作用により分解除去する分解除去層を用いたことを特徴とする。さらに、本発明のマイクロアレイチップは、このような分解除去層と基材との濡れ性の差を利用して固定化層を形成した場合と、分解除去層自体を固定化層として用いた場合とに分けることができる。以下、両方の場合を第３実施態様および第４実施態様とし、各態様について説明する。
【０２５０】
１．第３実施態様
第３実施態様のマイクロアレイチップは、基材と異なる濡れ性を示し、さらに、エネルギー照射に伴う光触媒の作用により分解除去される分解除去層を利用して製造されたものである。
【０２５１】
このような第３実施態様のマイクロアレイチップは、基材と、前記基材上にパターン状に形成され、エネルギー照射に伴う光触媒の作用により分解除去する分解除去層と、前記分解除去層上または前記分解除去層が分解除去され露出している基材上に形成され、生体高分子を固定化するための固定化層と、前記固定化層上に固定化され、ターゲットとなる生体高分子と特異的に結合する特異性生体高分子とを有し、前記分解除去層および前記基は、液体に対する接触角が異なることを特徴とするものである。
【０２５２】
このような本態様のマイクロアレイチップについて図面を用いて説明する。
【０２５３】
図４は、このような本態様におけるマイクロアレイチップの一例を示している。まず、基材４上には分解除去層２１がパターン状に形成されている。ここで、本態様においては、分解除去層と基材との濡れ性が異なるため、基材上に分解除去層がパターン状に形成されていることにより、濡れ性の違いによるパターンが形成されていることとなる。この濡れ性の違いによるパターンを利用して固定化層１０が分解除去層が分解除去され露出している基材４上に形成されている。なお、図４には、基材上に固定化層が形成されている場合を示したが、基材と分解除去層との濡れ性の関係から、分解除去層上に形成されている場合であってもよい。さらに、固定化層１０上には特異性生体高分子１１が固定されている。
【０２５４】
このような本実施態様におけるマイクロアレイチップにおいては、固定化層がパターン状に形成され、その上に特異性生体高分子が配置されている。さらに、固定化層は、分解除去層と基材との濡れ性の差を利用してパターン状に形成されていることから、固定化層が形成されていない領域は、撥水性の領域となっている。従って、例えば、インクジェット方式等で、固定化層上に特異性生態高分子が配置された場合に、特性生体高分子が固定化層の領域を越えて濡れ広がることが無く、特異性生体高分子の形状を揃えることが可能であり、最終的な検出感度を向上させることができる利点を有している。
【０２５５】
なお、本態様における、基材、分解除去層、固定化層、特異性生体高分子等は上述した「Ｉ．マイクロアレイチップの製造方法」で説明したものと同様であるので、ここでの説明は省略する。
【０２５６】
２．第４実施態様
次に、第４実施態様について説明する。第４実施態様は、エネルギー照射に伴う光触媒の作用により分解除去され、かつ固定化層として機能する分解除去層を用いて形成されたマイクロアレイチップである。
【０２５７】
このような本態様におけるマイクロアレイチップは、基材と、前記基材上にパターン状に形成され、エネルギー照射に伴う光触媒の作用により分解除去し、固定化層として機能する分解除去層と、前記分解除去層上に固定化され、ターゲットとなる生体高分子と特異的に結合する特異性生体高分子とを有することを特徴とするものである。
【０２５８】
さらに、本態様のマイクロアレイチップについて図面を用いて説明する。
【０２５９】
図５は、本態様のマイクロアレイチップの一例を示している。まず、基材４と、この基材４上には、パターン状に分解除去層２１が形成されている。本態様においては、この分解除去層５が固定化層としての機能を有していることから、この分解除去層５上に、特異性生体高分子１１を付着させ、固定させることにより、マイクロアレイチップとすることができる。
【０２６０】
本態様によれば、エネルギー照射のみで容易にパターニングすることが可能な分解除去層を用い、かつ分解除去層を固定化層として用いることにより、固定化層のパターニングに要する手間を大幅に省略することができ、製造工程を簡略化することができるのである。また、材料面においても無駄を少なくすることができるため、コスト的にも有利である。
【０２６１】
なお、本態様における、基材、分解除去層、特異性生体高分子等は上述した「Ｉ．マイクロアレイチップの製造方法」で説明したものと同様であるので、ここでの説明は省略する。
【０２６２】
ＩＩＩ．再生方法
本発明においては、上述したマイクロアレイチップを用いて検出等に使用した後、上述した光触媒処理層側基板を介して、上記固定化層および特異性生体高分子が付着した部分のみパターン状にエネルギー照射を行なうことにより再生することができる。
【０２６３】
本発明のマイクロアレイチップは、このように再生可能であることから、複数回にわたって使用することが可能であり、最終的なコストを低減させることができるという利点を有するものである。
【０２６４】
図６は、本発明のマイクロアレイチップの再生方法の一例を説明するためのものである。図６（ａ）は、生体高分子の検出等の使用が終わった後のマイクロアレイチップを示すものである。本発明のマイクロアレイチップの再生方法においては、図６（ｂ）に示すように、このような使用済みのマイクロアレイチップに対して、その固定化層１０および特異性生体高分子１１が配置されている部分のみ、エネルギーが照射されるように、紫外光等のエネルギーをフォトマスク７を介し、かつ、光触媒処理層側基板３を、光触媒処理層２と特異性生体高分子１１とが向かい合うように配置し、エネルギーを照射する。これにより、固定化層１０および特異性生体高分子１１は、光触媒処理層２に含有されている光触媒の作用により分解される。一方、エネルギーが照射されない部分はそのまま撥水性領域として残存する。この場合の、光触媒処理層側基板とマイクロアレイチップとの間隙およびエネルギー照射方法等は、上述した第1実施態様における「Ｂ．パターン形成工程」の中に記載したものと同様である。また、必要であれば洗浄等を行うことにより、図６（ｃ）に示すように、濡れ性変化層５上にパターン状に親水性領域８が形成された基材４を得る。
【０２６５】
このような基材に対して、上述した図１（ｄ）および（ｅ）に示す工程を行なうことにより、マイクロチップアレイとして再度利用することが可能となる。
【０２６６】
図７は、本発明のマイクロチップアレイの再生方法の他の例を示すものである。このマイクロチップアレイには、基材４と濡れ性変化層５との間にマイクロアレイチップ側遮光部３３が形成されており、このマイクロアレイチップ側遮光部３３は固定化層が形成される部分を区切る位置に形成されている（図７（ａ）参照）。このようなマイクロアレイチップを再生する場合は、図７（ｂ）に示すように、光触媒処理層側基板３を、光触媒処理層２と特異性生体高分子１１とが向かい合うように配置し、基材４の濡れ性変化層５が形成されていない面側からエネルギーを全面照射することにより、マイクロアレイチップ側遮光部３３が形成されていない部分、すなわち固定化層１０および特異性生体高分子１１が付着している部分のみエネルギーが照射されることになり、これらは光触媒処理層２に含有されている光触媒の作用により除去され、図７（ｃ）に示すように濡れ性変化層５にパターン状に親水性領域８が形成された基材４を得る。この際の、光触媒処理層側基板の配置の仕方、エネルギー照射に関することは、上述した第１実施態様における「Ｂ．パターン形成方法」の中に記載したことと同様であるので、ここでの記載は省略する。
【０２６７】
また、本態様におけるマイクロアレイチップ側遮光部とは、エネルギー照射の際にエネルギーの透過を妨げる役割を担う部材であり、上述した光触媒処理層側遮光層と同様のものを用いることが可能である。従って、マイクロアレイチップ側遮光部に関することは、上述した光触媒処理層側遮光部と同様であるので、ここでの説明は省略する。
【０２６８】
この場合も、上述した例と同様に、この基材に対して、上述した図１（ｄ）および（ｅ）に示す工程を行なうことにより、マイクロチップアレイとして再度利用することが可能となる。
【０２６９】
このような再生方法として、図６および図７には、濡れ性変化層を用いて形成されたマイクロアレイチップの場合について示したが、分解除去層を用いて形成した場合のマイクロアレイチップの場合にも同様の再生方法により、再生させることができる。
【０２７０】
なお、上記マイクロアレイチップの再生方法における、基材、濡れ性変化層、光触媒処理層側基板、固定化層、特異性生体高分子等は上述した「Ｉ．マイクロアレイチップの製造方法」で説明したものと同様であるので、ここでの説明は省略する。
【０２７１】
なお、本発明は、上記実施形態に限定されるものではない。上記実施形態は例示であり、本発明の特許請求の範囲に記載された技術的思想と実質的に同一な構成を有し、同様な作用効果を奏するものは、いかなるものであっても本発明の技術的範囲に包含される。
【０２７２】
【実施例】
以下に実施例を示し、本発明をさらに説明する。
［実施例１］
１．光触媒処理層側基板の形成
トリメトキシメチルシラン（ＧＥ東芝シリコーン（株）製、ＴＳＬ８１１３）５ｇと０．５規定塩酸を２．５ｇを混合し、８時間攪拌した。これをイソプロピルアルコールにより１０倍に希釈しプライマー層用組成物とした。
【０２７３】
上記プライマー層用組成物をフォトマスク基板上にスピンコーターにより塗布し、１５０℃で１０分間の乾燥処理を行うことにより、透明なプライマー層（厚み０．２μｍ）を形成した。
【０２７４】
次に、イソプロピルアルコール３０ｇとトリメトキシメチルシラン（ＧＥ東芝シリコーン（株）製、ＴＳＬ８１１３）３ｇと光触媒無機コーティング剤であるＳＴ−Ｋ０３（石原産業（株）製）２０ｇとを混合し、１００℃で２０分間撹拌した。これをイソプロピルアルコールにより３倍に希釈し光触媒処理層用組成物とした。
【０２７５】
上記光触媒処理層用組成物をプライマー層が形成されたフォトマスク基板上にスピンコーターにより塗布し、１５０℃で１０分間の乾燥処理を行うことにより、透明な光触媒処理層（厚み０．１５μｍ）を形成した。
【０２７６】
２．マイクロアレイチップ用基材の調整
イソプロピルアルコール３０ｇとフルオロアルキルシラン（ＧＥ東芝シリコーン（株）製）０．４ｇとトリメトキシメチルシラン（ＧＥ東芝シリコーン（株）製、ＴＳＬ８１１３）３ｇと０．５規定塩酸２．５ｇを混合し８時間攪拌した。これをイソプロピルアルコールにより１００倍に希釈し濡れ性変化層用組成物とした。
【０２７７】
上記濡れ性変化層用組成物をガラス基板上にスピンコーターにより塗布し、１５０℃で１０分間の乾燥処理を行うことにより、透明な濡れ性変化層（厚み０．１μｍ）を形成した。
【０２７８】
３．露光による親水性領域の形成の確認
光触媒処理層側基板と濡れ性変化層とを１００μｍのギャップを設けて対向させて、フォトマスク側から超高圧水銀灯（波長３６５ｎｍ）により４０ｍＷ／ｃｍ²の照度で６０秒間露光し、濡れ性変化層上の固定化層が形成される領域を親水性領域とした。
【０２７９】
このとき、未露光部からなる撥液性領域及び親水性領域と表面張力４０ｍＮ／ｍの濡れ指数標準液（純正化学株式会社製）との接触角を接触角測定器（協和界面科学（株）製ＣＡ−Ｚ型）を用いて測定（マイクロシリンジから液滴を滴下して３０秒後）した結果、それぞれ、７０度と７度であった。
【０２８０】
４．固定化層の形成
上記基板をポリL−リシンの３％水溶液に浸漬し引き上げることにより、親水性領域のみに固定化層を形成した。
【０２８１】
５．マイクロアレイチップの作製
上記固定化層上に各種DNA溶液をインクジェット装置にて吐出し、固定させることによりマイクロアレイチップを作製した。
【０２８２】
６．再生方法
１の光触媒処理層側基板とマイクロアレイチップとをアライメントをとり１００μｍのギャップを設けて対向させて、光触媒処理層側基板から超高圧水銀灯（波長３６５ｎｍ）により４０ｍＷ／ｃｍ²の照度で３００秒間露光し、再生した。
[実施例２]
１．光触媒処理層側基板の形成
トリメトキシメチルシラン（ＧＥ東芝シリコーン（株）製、ＴＳＬ８１１３）５ｇと０．５規定塩酸を２．５ｇを混合し、８時間攪拌した。これをイソプロピルアルコールにより１０倍に希釈しプライマー層用組成物とした。
【０２８３】
上記プライマー層用組成物をフォトマスク基板上にスピンコーターにより塗布し、１５０℃で１０分間の乾燥処理を行うことにより、透明なプライマー層（厚み０．２μｍ）を形成した。
【０２８４】
次に、イソプロピルアルコール３０ｇとトリメトキシメチルシラン（ＧＥ東芝シリコーン（株）製、ＴＳＬ８１１３）３ｇと光触媒無機コーティング剤であるＳＴ−Ｋ０３（石原産業（株）製）２０ｇとを混合し、１００℃で２０分間撹拌した。これをイソプロピルアルコールにより３倍に希釈し光触媒処理層用組成物とした。
【０２８５】
上記光触媒処理層用組成物をプライマー層が形成されたフォトマスク基板上にスピンコーターにより塗布し、１５０℃で１０分間の乾燥処理を行うことにより、透明な光触媒処理層（厚み０．１５μｍ）を形成した。
【０２８６】
２．固定化層の形成
ガラス基板をポリL−リシンの３％水溶液に浸漬し引き上げることにより、固定化層を形成した。
【０２８７】
３．露光による固定化層のパターニング
光触媒処理層側基板と固定化層とを１００μｍのギャップを設けて対向させて、フォトマスク側から超高圧水銀灯（波長３６５ｎｍ）により４０ｍＷ／ｃｍ²の照度で１８０秒間露光し、固定化層をパターニングした。
【０２８８】
４．マイクロアレイチップの作製
上記固定化層上に各種DNA溶液をインクジェット装置にて吐出し、固定させることによりマイクロアレイチップを作製した。
【０２８９】
６．再生方法
光触媒処理層側基板とマイクロアレイチップとを１００μｍのギャップを設けて対向させて、光触媒処理層側基板から超高圧水銀灯（波長３６５ｎｍ）により４０ｍＷ／ｃｍ²の照度で３００秒間露光し、再生した。
【０２９０】
【発明の効果】
本発明によれば、エネルギー照射後、マイクロアレイチップ用基材から光触媒処理層側基板を取り外すので、マイクロアレイチップ自体には光触媒処理層が含まれることがなく、従って、マイクロアレイチップに光触媒の半永久的な作用が及ぶ心配がない。さらに、光触媒処理層と特性変化層との間隔が、上述した範囲内であるので、効率よく、精度の良好な特性の違いによるパターンを形成することができる。さらに、固定化層はエネルギーのパターン照射により形成された特性変化層の変化のパターンを利用して形成されるので、高精細なパターンとすることができる。従って、固定化層上に特異性生体高分子を固定させることにより高精細なマイクロアレイチップとすることができる。
【図面の簡単な説明】
【図１】本発明のマイクロアレイチップの製造方法の一例を示す工程図である。
【図２】本発明のマイクロアレイチップの製造方法の他の例を示す工程図である。
【図３】本発明のマイクロアレイチップの製造方法の他の例を示す工程図である。
【図４】本発明のマイクロアレイチップの一例を示す概略断面図である。
【図５】本発明のマイクロアレイチップの他の例を示す概略断面図である。
【図６】本発明のマイクロアレイチップの再生方法の一例を説明するための工程図である。
【図７】本発明のマイクロアレイチップの再生方法の他の例を説明するための工程図である。
【図８】本発明の光触媒処理層側基板の一例を示す概略断面図である。
【図９】本発明の光触媒処理層側基板の他の例を示す概略断面図である。
【図１０】本発明の光触媒処理層側基板の他の例を示す概略断面図である。
【図１１】本発明の光触媒処理層側基板の他の例を示す概略断面図である。
【符号の説明】
１ … 基板
２ … 光触媒処理層
３ … 光触媒処理層側基板
４ … 基材
５ … 濡れ性変化層
６ … マイクロアレイチップ用基材
７ … フォトマスク
８ … 親水性領域
９ … 撥水性領域
１０ … 固定化層
１１ … 特異性生体高分子[0001]
BACKGROUND OF THE INVENTION
The present invention relates to a method for producing a microarray chip in which a large number of specific biopolymers that specifically bind to specific DNA or protein are arranged.
[0002]
[Prior art]
In the field of molecular biology, microarray chips in which biopolymers such as DNA and proteins are immobilized at a high density have attracted rapid attention in recent years. In an experiment using a microarray chip, a large amount of a support (chip) on which a large number of samples (samples) are immobilized is replicated in large quantities, and the sample on each chip is reacted with a sample (target) for investigation control. . It is possible to detect a difference in reaction between samples on one chip by one experiment and a difference in reaction between the same sample set among a plurality of chips by repeating an experiment with different conditions.
[0003]
As a method for producing such a microarray chip, first, a method developed by Affimetrix can be mentioned. This method synthesizes oligonucleotides at high density for every several tens of μm rectangles on a substrate using protective groups that are selectively removed by light irradiation and photolithography technology used in semiconductor manufacturing. . The microarray chip obtained by this method is currently the only standardized standard in the world, but the price is very high and there is a problem that only some researchers can use it sufficiently. there were.
[0004]
On the other hand, as another method for manufacturing a microarray chip, a method developed by P. Brown et al. This method is very simple compared to the Affimetrix method described above, in which pre-prepared DNA is dispensed into 96 or 384-well microplates that have been chemically treated on the surface, resulting in high density spots. This is a method of spotting with high density on a slide glass with a possible robot. According to this method, the manufacturing cost is low, and custom chips for many researchers can be created. Therefore, this method is used in many research institutions.
[0005]
Such a Stanford method is usually produced by applying a fixing agent for immobilizing DNA on a glass substrate and spotting the target DNA at a high density thereon. However, in the case where the surface of the immobilizing agent is hydrophilic, there is a possibility that the spotted DNA is diffused and the adjacent spots come into contact with each other.
[0006]
In order to solve such problems, the present inventors have studied a method for manufacturing a microarray chip using a pattern formed using a substance whose wettability is changed by the action of a photocatalyst. . This makes it possible to suppress the occurrence of the above-described problems. However, the conventional microarray chip manufacturing method based on the action of the photocatalyst has a configuration in which the photocatalyst is included in the manufactured microarray chip itself, so that the action of the photocatalyst contained therein is semipermanently microarray chip. In some cases, it may have a problem that it may affect.
[0007]
[Problems to be solved by the invention]
The present invention has been made in view of the above problems, and since it does not contain a photocatalyst inside, there is no fear of being semipermanently affected by the photocatalyst, and DNA can be concentrated at a high density regardless of the type of immobilizing agent. The main object of the present invention is to provide a method for manufacturing a microarray chip that does not have a problem of spotted DNA spreading even when spotted.
[0008]
[Means for Solving the Problems]
In order to achieve the above object, the present invention provides a microarray chip substrate for adjusting a microarray chip substrate having a characteristic change layer whose characteristics are changed by the action of a photocatalyst associated with energy irradiation. Using the material adjustment step and a photocatalyst treatment layer side substrate on which a photocatalyst treatment layer containing at least a photocatalyst is formed on the substrate, the microarray chip substrate and the photocatalyst treatment layer side substrate are combined with the photocatalyst treatment layer and the photocatalyst treatment layer. A pattern forming step of forming a pattern due to a difference in characteristics on the surface of the characteristic change layer by irradiating energy from a predetermined direction after being arranged with the characteristic change layers facing each other and with a gap of 200 μm or less; Removing the photocatalyst treatment layer side substrate from the microarray chip substrate; and It has at least an immobilization layer forming step for forming an immobilization layer and a specific biopolymer immobilization step for immobilizing a specific biopolymer on the immobilization layer according to a pattern depending on a difference in characteristics of the change layer. A method of manufacturing a microarray chip is provided.
[0009]
According to the present invention, after the energy irradiation, the photocatalyst processing layer side substrate is removed from the microarray chip substrate, so that the microarray chip itself does not include the photocatalyst processing layer. There is no worry about the effect. Furthermore, since the distance between the photocatalyst treatment layer and the characteristic change layer is within the above-described range, a pattern can be formed efficiently and with a good difference in characteristics. Furthermore, since the immobilization layer is formed using the change pattern of the characteristic change layer formed by energy pattern irradiation, a high-definition pattern can be obtained. Therefore, a high-definition microarray chip can be obtained by immobilizing a specific biopolymer on the immobilization layer.
[0010]
In the present invention, as described in claim 2, a microarray chip substrate adjusting step of adjusting a microarray chip substrate having a wettability changing layer in which wettability is changed by the action of a photocatalyst accompanying energy irradiation; A substrate having a photocatalyst treatment layer on which a photocatalyst treatment layer containing at least a photocatalyst is formed, the substrate for microarray chip and the photocatalyst treatment layer side substrate, the photocatalyst treatment layer and the wettability changing layer; Forming a pattern due to a difference in wettability on the surface of the wettability changing layer by irradiating energy from a predetermined direction after being arranged with a gap of 200 μm or less so as to face each other, Removal step of removing the photocatalyst processing layer side substrate from the microarray chip substrate, and the wettability An immobilized layer forming step of forming an immobilized layer on the wettability changing layer according to the pattern of the immobilized layer, and a specific biopolymer immobilizing step of immobilizing a specific biopolymer on the immobilized layer A method for manufacturing a microarray chip is provided.
[0011]
According to the present invention, after the energy irradiation, the photocatalyst processing layer side substrate is removed from the microarray chip substrate, so that the microarray chip itself does not include the photocatalyst processing layer. There is no worry about the effect. Furthermore, since the distance between the photocatalyst treatment layer and the wettability changing layer is within the above-described range, a pattern due to a difference in wettability with good accuracy can be formed efficiently. Further, since the immobilization layer is formed using a pattern due to such a difference in wettability, the region other than the region where the immobilization layer is formed exhibits water repellency. Therefore, since the specific biopolymer spotted on the immobilization layer does not spread over the immobilization layer region, there is no problem that adjacent spots come into contact with each other.
[0012]
In the invention described in claim 2, as described in claim 3, the wettability changing layer is preferably a layer containing organopolysiloxane, and in particular, as described in claim 4. In addition, the organopolysiloxane is preferably a polysiloxane containing a fluoroalkyl group. This is because such a wettability changing layer can obtain a significant change in wettability by energy irradiation in a state where the photocatalyst processing layer is in contact.
[0013]
In the invention described in claim 3 or 4, as described in claim 5, the organopolysiloxane is Y_nSiX_(4-n)Wherein Y represents an alkyl group, a fluoroalkyl group, a vinyl group, an amino group, a phenyl group or an epoxy group, X represents an alkoxyl group or a halogen, and n is an integer from 0 to 3. It is preferable that it is the organopolysiloxane which is a 1 type, or 2 or more types of hydrolyzed condensate or cohydrolyzed condensate of the silicon compound. This is because by forming a wettability changing layer using such an organopolysiloxane as a material, a microarray chip having a wettability pattern having a large difference in wettability can be obtained.
[0014]
In the invention described in any one of claims 2 to 5, as described in claim 6, the substrate for microarray chip is formed on the substrate and the substrate. The wettability changing layer is preferably at least. This is because, for example, when the wettability changing layer does not have self-supporting property, the strength can be maintained by forming the wettability changing layer on the substrate.
[0015]
In the invention described in claim 2, as described in claim 7, the base material for microarray chip may comprise a wettability changing layer having self-supporting property. If the wettability changing layer has a self-supporting property, it is not necessary to use a substrate or the like, and a microarray chip can be manufactured at low cost.
[0016]
In the present invention, as described in claim 8, a decomposition removal layer is formed on the substrate by being decomposed and removed by the action of a photocatalyst accompanying energy irradiation and having a contact angle with a liquid different from that of the substrate. A microarray chip base material adjusting step for adjusting the microarray chip base material, and a photocatalyst processing layer side substrate on which a photocatalyst processing layer containing at least a photocatalyst is formed on the substrate, and the microarray chip base material and After disposing the photocatalyst treatment layer side substrate so that the photocatalyst treatment layer and the decomposition / removal layer face each other with a gap of 200 μm or less, the decomposition / removal layer is irradiated with energy from a predetermined direction. A pattern forming step of forming a pattern, and removing the photocatalyst processing layer side substrate from the microarray chip substrate. In accordance with the pattern of the decomposition / removal layer, an immobilization layer forming step of forming an immobilization layer on the decomposition / removal layer or the substrate, and immobilizing the specific biopolymer on the immobilization layer. There is provided a method for producing a microarray chip, comprising at least a specific biopolymer immobilization step.
[0017]
According to the present invention, since the photocatalyst processing layer side substrate is removed from the microarray chip substrate after energy irradiation, the photocatalyst processing layer is not included in the microarray chip itself, and therefore the semi-permanent action of the photocatalyst is microarray. There is no worry about the tip. Furthermore, since the wettability of the decomposition removal layer and the substrate is different, a pattern due to the difference in wettability can be formed on the substrate by forming the decomposition removal layer in a pattern. Since the immobilization layer is formed using a pattern, water repellency is exhibited except in the region where the immobilization layer is formed. Therefore, since the specific biopolymer spotted on the immobilization layer does not spread over the immobilization layer region, there is no problem that adjacent spots come into contact with each other.
[0018]
Furthermore, in the present invention, as described in claim 9, a decomposition removal layer that is decomposed and removed by the action of a photocatalyst accompanying energy irradiation and functions as an immobilization layer is formed on a substrate, and is used for a microarray chip. Using the substrate adjustment step for microarray chip for adjusting the substrate and the photocatalyst treatment layer side substrate on which the photocatalyst treatment layer containing at least the photocatalyst is formed on the substrate, the substrate for microarray chip and the photocatalyst treatment layer side After disposing the substrate so that the photocatalyst treatment layer and the decomposition / removal layer face each other and with a gap of 200 μm or less, the decomposition / removal layer is formed in a pattern by irradiating energy from a predetermined direction. Pattern forming step and removing step of removing the photocatalyst processing layer side substrate from the microarray chip substrate , To provide a method of manufacturing a microarray chip, characterized in that it comprises at least a specific biopolymer fixing step of fixing the specificity biopolymers to the decomposition removal layer.
[0019]
According to the present invention, since the photocatalyst processing layer side substrate is removed from the microarray chip substrate after energy irradiation, the photocatalyst processing layer is not included in the microarray chip itself, and therefore the semi-permanent action of the photocatalyst is microarray. There is no worry about the tip. Furthermore, since the decomposition removal layer in the present invention has a function as an immobilization layer, the immobilization layer is also formed in a pattern by forming the decomposition removal layer in a pattern, thereby simplifying the manufacturing process. And a microarray chip can be manufactured efficiently.
[0020]
In the invention described in claim 8 or claim 9, as described in claim 10, the decomposition / removal layer is a self-assembled monolayer, a Langmuir-Blodgett film, or an alternating adsorption film. It is preferable that This is because these materials are decomposed and removed by the action of the photocatalyst in the photocatalyst treatment layer and exhibit various functions.
[0021]
In the invention described in any one of claims 1 to 10, as described in claim 11, the photocatalyst processing layer side substrate includes a substrate and a pattern on the substrate. It is preferable to consist of the formed photocatalyst processing layer. Thus, by forming the photocatalyst treatment layer in a pattern, it is possible to form patterns having different characteristics in the wettability changing layer or the decomposition removal layer without using a photomask. In addition, since wettability changes or decomposes and removes only the surface corresponding to the photocatalyst treatment layer, the energy to be irradiated is not particularly limited to parallel energy, and the energy irradiation direction is also particularly limited. This is because it has the advantage that the type of energy source and the degree of freedom in arrangement are greatly increased.
[0022]
In the invention described in any one of claims 1 to 10, as described in claim 12, the photocatalyst processing layer side substrate is formed on the substrate and the substrate. It is preferable that the photocatalyst treatment layer and the photocatalyst treatment layer side light-shielding portion formed in a pattern are formed, and the energy irradiation in the pattern formation step is preferably performed from the photocatalyst treatment layer side substrate. By having the photocatalyst treatment layer side light-shielding portion on the photocatalyst treatment layer side substrate in this way, it is not necessary to use a photomask or the like when irradiating energy, so that alignment with the photomask is unnecessary, and the process is simplified. This is because it becomes possible.
[0023]
In the invention described in claim 12, as described in claim 13, the photocatalyst processing layer side substrate has a photocatalyst processing layer formed on the substrate, and the photocatalyst processing layer is formed on the photocatalyst processing layer. The side light-shielding portion may be formed in a pattern shape, or the photocatalyst treatment layer side substrate may be formed in a pattern shape on the substrate. The photocatalyst treatment layer may be formed on the photocatalyst treatment layer.
[0024]
Since the photocatalyst treatment layer side light-shielding portion is disposed in the vicinity of the portion where the photocatalyst treatment layer and the characteristic change layer are located with a gap, the influence of energy scattering in the substrate or the like can be reduced. Therefore, the pattern irradiation of energy can be performed very accurately. Further, when the photocatalyst treatment layer side light shielding portion is formed on the photocatalyst treatment layer, it has an advantage that it can be used as a spacer when arranging the photocatalyst treatment layer and the characteristic change layer in the pattern forming step. .
[0025]
In the invention described in claim 14, as described in claim 15, the photocatalyst processing layer side substrate is formed on a transparent substrate in a pattern shape on the photocatalyst processing layer side light shielding portion. It is preferable that the photocatalyst processing layer is formed via This is because patterning of the wettability changing layer and the decomposition removal layer proceeds with high sensitivity, and as a result, a high-resolution pattern can be obtained.
[0026]
In the invention described in any one of claims 1 to 10, as described in claim 16, in the photocatalyst processing layer side substrate, a thickness of 0. Spacers in the range of 2 μm to 10 μm are formed in a pattern, and it is preferable to irradiate the spacer with the spacer and the property change layer, the wettability change layer, or the decomposition / removal layer for energy irradiation. In this way, spacers are provided in a pattern on the photocatalyst treatment layer, and energy irradiation is performed so that the spacer is brought into contact with the property change layer or the like, so that the distance between the photocatalyst treatment layer and the property change layer or the like is 0.2 μm to It can be easily maintained within a range of 10 μm. In addition, since the photocatalyst processing layer is covered with the spacer in the portion where the spacer is formed, even if this portion is irradiated with energy, the property does not change on the property change layer, for example. Therefore, it is possible to form a pattern due to a difference in characteristics in the characteristic change layer or the like with the same pattern as the pattern in which the spacer is formed.
[0027]
In the invention described in claim 16, as described in claim 17, it is preferable that the spacer is a photocatalyst treatment layer side light shielding portion formed of a light shielding material. Since the spacer is the photocatalyst processing layer side light shielding part, it is possible to form a higher definition pattern by irradiating energy while the photocatalyst processing layer side light shielding part is in close contact with the characteristic change layer or the like. Because.
[0028]
In the invention described in any one of claims 1 to 17, as described in claim 18, the photocatalyst treatment layer is preferably a layer made of a photocatalyst. This is because if the photocatalyst treatment layer is a layer composed only of a photocatalyst, it is possible to improve the efficiency of changing the characteristics of the characteristic change layer and the like, and the microarray chip can be manufactured efficiently.
[0029]
In the invention described in claim 18, as described in claim 19, it is preferable that the photocatalyst processing layer is a layer formed by forming a photocatalyst on a substrate by a vacuum film forming method. By forming the photocatalyst treatment layer by the vacuum film formation method in this way, it is possible to obtain a homogeneous photocatalyst treatment layer having a uniform film thickness with less surface irregularities, and due to the difference in properties to the property change layer, etc. This is because the pattern can be formed uniformly and with high efficiency.
[0030]
In the invention described in any one of claims 1 to 17, as described in claim 20, the photocatalyst treatment layer may be a layer having a photocatalyst and a binder. This is because the photocatalyst treatment layer can be formed relatively easily by using the binder in this manner.
[0031]
In the invention described in any one of claims 1 to 20, as described in claim 21, the photocatalyst is composed of titanium oxide (TiO 2).₂), Zinc oxide (ZnO), tin oxide (SnO)₂), Strontium titanate (SrTiO)₃), Tungsten oxide (WO₃), Bismuth oxide (Bi₂O₃), And iron oxide (Fe₂O₃It is preferable that it is 1 type, or 2 or more types of substances selected from these, Among these, as described in Claim 22, the said photocatalyst is a titanium oxide (TiO2).₂) Is preferable. This is because titanium dioxide has a high band gap energy and is effective as a photocatalyst, and is chemically stable, non-toxic and easily available.
[0032]
In the invention described in any one of claims 1 to 22, as described in claim 23, when irradiating energy in the pattern forming step, the photocatalyst treatment layer and The distance from the property change layer, the wettability change layer, or the decomposition / removal layer is preferably in the range of 0.2 μm to 10 μm. This is because the distance between the photocatalyst treatment layer and the characteristic change layer is within a range of 0.2 μm to 10 μm, and therefore, a pattern due to a difference in characteristics can be formed in the characteristic change layer by short-time energy irradiation. .
[0033]
In the invention described in any one of claims 1 to 23, as described in claim 24, it is preferable that the energy irradiation is performed while heating the photocatalyst treatment layer. This is because by heating the photocatalyst, the sensitivity of the photocatalyst is improved, and a pattern can be efficiently formed in the characteristic change layer or the like.
[0034]
In the invention described in any one of claims 1 to 24, as described in claim 25, the immobilization layer may be formed of a cationic substance. preferable. This is because a substance having a normal cationic property can electrostatically immobilize an anionic biopolymer such as an oligonucleotide.
[0035]
In the invention described in any one of claims 1 to 25, as described in claim 26, the specific biopolymer specifically binds to a target nucleic acid. It is preferable that the probe DNA. This is because a detection method using probe DNA and utilizing hybridization with a fluorescently labeled target DNA has been established.
[0036]
In the invention described in any one of claims 1 to 26, as described in claim 27, a portion where the fixing layer is formed is separated on the base material. It is preferable that a microarray chip side light-shielding portion is formed. This is because the microarray chip regeneration method described later can be used without a mask.
[0037]
In the present invention, as described in claim 28, a base material, a decomposition removal layer formed in a pattern on the base material and decomposed and removed by the action of a photocatalyst associated with energy irradiation, and the decomposition removal layer An upper layer formed on the substrate on which the decomposition / removal layer is decomposed and removed and is exposed, and an immobilization layer for immobilizing a biopolymer, and a living body height which is immobilized on the immobilization layer and becomes a target There is provided a microarray chip having a specific biopolymer that specifically binds to a molecule, wherein the decomposition removal layer and the substrate have different contact angles with respect to a liquid.
[0038]
In the present invention, since the wettability is different between the decomposition removal layer and the substrate, the immobilization layer is formed using such a difference in wettability. Therefore, in order to show water repellency except in the region where the immobilization layer is formed, the specific biopolymer spotted on the immobilization layer does not spread out over the immobilization layer, and adjacent to each other. There is no problem of spot contact.
[0039]
Furthermore, in the present invention, as described in claim 29, the substrate is formed in a pattern on the substrate, and is decomposed and removed by the action of a photocatalyst associated with energy irradiation, and functions as an immobilization layer. There is provided a microarray chip comprising: a decomposition / removal layer to be immobilized; and a specific biopolymer that is immobilized on the decomposition / removal layer and specifically binds to a target biopolymer.
[0040]
According to the present invention, since the decomposition removal layer that is decomposed and removed by the action of the photocatalyst accompanying the energy irradiation and functions as the immobilization layer is used, the immobilization layer is separately formed by forming the decomposition removal layer in a pattern shape. There is no need to provide. Accordingly, since the decomposition / removal layer having a function as an immobilization layer can be formed in a pattern only by energy irradiation, the manufacturing process can be greatly simplified, and a microarray chip advantageous in terms of cost can be obtained.
[0041]
In the invention described in claim 28 or claim 29, as described in claim 30, the decomposition / removal layer is a self-assembled monolayer, a Langmuir-Blodgett film, or an alternating adsorption film. It is preferable that
[0042]
This is because, by forming the decomposition / removal layer by such a manufacturing method, a thin film having a relatively high strength that can be decomposed by the action of the photocatalyst can be formed. In addition, by selecting the material of the thin film, it is possible to form a difference in good wettability from the base material exposed when the decomposition removal layer is decomposed and removed, and the decomposition removal layer can be used as an immobilization layer. This is because the immobilization layer can be easily patterned by selecting a material that functions in this manner.
[0043]
In the invention described in any one of claims 28 to 30, as described in claim 31, the immobilization layer may be formed of a cationic substance. preferable. This is because a substance having a normal cationic property can electrostatically immobilize an anionic biopolymer such as an oligonucleotide.
[0044]
Furthermore, in the present invention, at least a photocatalyst is contained on the substrate after using the microarray chip manufactured by the method for manufacturing a microarray chip according to any one of claims 1 to 27. There is provided a method for regenerating a microarray chip, wherein energy is irradiated in a pattern only to a portion where the specific biopolymer is attached via a photocatalyst treatment layer side substrate on which the photocatalyst treatment layer is formed.
[0045]
In the present invention, by irradiating only the region in which the immobilization layer and the specific biopolymer are formed through the photocatalyst treatment layer containing the photocatalyst, the immobilization agent and The specific biopolymer can be removed, and it can be reused as a microarray chip by forming the immobilization layer and the specific biopolymer again.
[0046]
DETAILED DESCRIPTION OF THE INVENTION
Hereinafter, the microarray chip of the present invention and the manufacturing method thereof will be described. First, the manufacturing method of the microarray of this invention is demonstrated in detail.
[0047]
I. Manufacturing method of microarray chip
The microarray chip manufacturing method of the present invention includes a microarray chip base material adjusting step of adjusting a microarray chip base material having a characteristic change layer whose characteristics are changed by the action of a photocatalyst accompanying energy irradiation, and at least a photocatalyst on the substrate. Using the photocatalyst processing layer-side substrate on which the photocatalyst processing layer is formed, the microarray chip substrate and the photocatalyst processing layer side substrate are 200 μm so that the photocatalyst processing layer and the property change layer face each other. A pattern forming step of forming a pattern due to a difference in characteristics on the surface of the characteristic change layer by irradiating energy from a predetermined direction after being arranged with a gap between the following, and the photocatalytic treatment from the substrate for the microarray chip The difference between the removal process of removing the layer side substrate and the characteristics change layer According to the pattern, at least an immobilization layer forming step for forming an immobilization layer in a pattern and a specific biopolymer immobilization step for immobilizing a specific biopolymer on the immobilization layer are provided. Is.
[0048]
Thus, in the microarray chip manufacturing method of the present invention, when patterning the characteristic change layer, the photocatalyst processing layer side substrate faces the substrate for microarray chip, so that the action of the photocatalyst in the photocatalyst processing layer is achieved. Since the characteristic of the portion irradiated with energy changes in the characteristic change layer, a pattern due to the difference in characteristic can be formed on the characteristic change layer. Furthermore, in the present invention, after such patterning is performed, the photocatalyst processing layer side substrate is removed and the microarray chip side base material is used as a microarray chip. Therefore, the obtained microarray chip includes a photocatalyst processing layer. Not. For this reason, it is possible to eliminate the possibility that the microarray chip is affected by the action of the photocatalyst.
[0049]
The term “microarray chip substrate” as used herein refers to a substrate in which a pattern due to a difference in characteristics is not formed on the property change layer, and the microarray chip substrate is irradiated with energy to have characteristics. A pattern of a characteristic change site is formed on the change layer, an immobilization layer is formed by a pattern based on the difference in the characteristics, and a microarray chip is obtained by immobilizing a specific biopolymer on the immobilization layer.
[0050]
The property changing layer used in the method of manufacturing the microarray chip of the present invention may be any layer as long as the property is changed by the action of the photocatalyst, such as a polyolefin such as polyethylene and polypropylene. By using a polymer material, etc., the part irradiated with energy is introduced with a polar group by the action of the photocatalyst, or the surface state becomes rough so that the adhesion to various substances is improved. This layer may be used as a characteristic change layer. Thus, by making the characteristic change layer an adhesive change layer in which the adhesiveness changes, it becomes possible to form a pattern with good adhesiveness by irradiation with an energy pattern. A microarray chip can be easily formed by forming an immobilization layer in such a site with good adhesion and further immobilizing a characteristic biopolymer on the immobilization layer.
[0051]
Further, in the present invention, such a characteristic change layer may be formed by a dry method, that is, a vacuum deposition method or the like, or by a wet method, that is, a method such as a spin coating method or a dip coating method. It may be formed.
[0052]
As described above, the characteristic change layer is not particularly limited as long as it has various characteristics that change due to the action of the photocatalyst. However, in the present invention, the characteristic change layer changes the wettability due to the action of the photocatalyst. There are two cases where the wettability changing layer in which a pattern due to the property is formed and the case where the property changing layer is a decomposition removing layer in which the pattern due to unevenness is formed by being decomposed and removed by the action of the photocatalyst. It is preferable because it brings out the sex.
[0053]
Therefore, hereinafter, the microarray chip manufacturing method of the present invention when a wettability changing layer or a decomposition removal layer is used as the characteristic changing layer will be described in detail as a first embodiment and a second embodiment, respectively.
[0054]
1. First embodiment
In the first embodiment, a wettability changing layer whose wettability changes due to the action of a photocatalyst accompanying energy irradiation is used as a property changing layer, and a pattern due to the difference in wettability is formed on the wettability changing layer. This is a method for manufacturing a microarray chip in which an immobilization layer is formed in a pattern by using it.
[0055]
Such a microarray chip manufacturing method in this embodiment includes a microarray chip base material adjusting step for adjusting a microarray chip base material having a wettability changing layer in which the wettability is changed by the action of a photocatalyst accompanying energy irradiation, and a substrate. Using a photocatalyst treatment layer side substrate on which a photocatalyst treatment layer containing at least a photocatalyst is formed, the photocatalyst treatment layer and the wettability changing layer face each other with the microarray chip substrate and the photocatalyst treatment layer side substrate. The pattern forming step of forming a pattern due to the difference in wettability on the surface of the wettability changing layer by irradiating energy from a predetermined direction after being arranged with a gap of 200 μm or less, and the microarray A removal step of removing the photocatalyst processing layer side substrate from the chip substrate; An immobilization layer forming step for forming an immobilization layer on the wettability change layer according to the pattern of the wettability change layer, and a specific biopolymer immobilization step for immobilizing a specific biopolymer on the immobilization layer And at least.
[0056]
According to the present invention, after forming a pattern due to the difference in wettability on the surface of the wettability changing layer by the action of the photocatalyst associated with energy irradiation, the microcatalyst processing layer side substrate is removed from the substrate for microarray chip. The structure of the chip does not include the photocatalyst processing layer, and therefore it is possible to eliminate the possibility that the microarray chip is affected by the action of the photocatalyst.
[0057]
The manufacturing method of the microarray chip in this embodiment will be described with reference to the drawings.
[0058]
FIG. 1 shows an example of a manufacturing method of such a microarray chip. In this example, first, a photocatalyst processing layer 2 having at least a photocatalyst is formed on a substrate 1 to prepare a photocatalyst processing layer side substrate 3. Further, separately from the photocatalyst processing layer side substrate 3, a base material 4 is prepared, and a wettability changing layer 5 in which the wettability is changed by the action of the photocatalyst accompanying energy irradiation is formed on the base material 4. A base material 6 is prepared (see FIG. 1A).
[0059]
Next, the photocatalyst processing layer side substrate 3 and the microarray chip substrate 6 are arranged so that the photocatalyst processing layer 2 and the wettability changing layer 5 face each other, and energy is irradiated through the photomask 7 (FIG. 1B). reference). Due to this energy irradiation, the wettability changing layer 5 of the irradiated portion changes its wettability in the direction in which the contact angle with the liquid decreases due to the action of the photocatalyst contained in the photocatalyst treatment layer 2, and is hydrophilic. Form a region. On the contrary, the non-irradiated region of energy is a water-repellent region without changing wettability.
[0060]
Further, after the energy irradiation, the photocatalyst processing layer side substrate 3 is removed from the microarray chip substrate 6. As a result, a wettability changing layer 5 in which a pattern due to a difference in wettability, that is, a hydrophilic region 8 and a water repellent region 9 is formed is obtained (see FIG. 1C).
[0061]
The fixed layer 10 is formed on the hydrophilic region 8 using a pattern due to the difference in wettability formed on the surface of the wettability changing layer 5 (see FIG. 1D). Next, a microarray chip is manufactured by attaching a specific biopolymer 11 on the immobilization layer 10 (see FIG. 1 (e)). At this time, the surface of the wettability changing layer 5 is a water-repellent region 9 except for the region where the immobilization layer 10 is formed. It doesn't spread over wet. Therefore, it is possible to prevent the adjacent specific biopolymers from being mixed with each other and to improve the final detection sensitivity.
[0062]
The manufacturing method of the microarray chip of this embodiment having such advantages will be described in detail for each step.
[0063]
A. Microarray chip substrate preparation process
First, the substrate adjustment process for microarray chips will be described. This step is a step of preparing a microarray chip substrate having a wettability changing layer in which the wettability changes due to the action of a photocatalyst accompanying energy irradiation.
[0064]
Such a microarray chip substrate has at least a wettability changing layer, and when the wettability changing layer has self-supporting property, it may be a microarray chip substrate made of a wettability changing layer. Moreover, when it is a wettability change layer with few self-supporting properties, the base material for microarray chips by which a wettability change layer is formed on a base material may be sufficient.
[0065]
In addition, having self-supporting property in this aspect means that it can exist in a tangible state without other supporting materials.
[0066]
Hereinafter, such a microarray chip substrate will be described.
[0067]
(Base material for microarray chip)
(1) Wetting change layer
First, the wettability changing layer will be described as a member constituting the microarray chip.
[0068]
The wettability changing layer in this embodiment is not particularly limited as long as the wettability of the surface is changed by the action of the photocatalyst, but in general, the wettability change is caused by the action of the photocatalyst accompanying the irradiation of energy. A layer whose wettability changes so that the contact angle with the liquid on the surface of the layer decreases is preferable.
[0069]
In this way, when the wettability changing layer whose wettability changes so that the contact angle with the liquid is reduced by energy irradiation, the photocatalyst processing layer side substrate is used as the base material for the microarray chip in the pattern forming step described later. This is because the wettability can be easily changed into a pattern by irradiating the pattern with energy. Thereby, a pattern comprising a hydrophilic region having a small contact angle with the liquid and a water repellent region having a large contact angle with the liquid is formed on the wettability changing layer.
[0070]
In addition, the hydrophilic region here is a region having a small contact angle with water as described above, and has good wettability with respect to an immobilization layer forming coating solution for forming an immobilization layer described later. Territory. Further, the water-repellent region is a region having a large contact angle with water, and refers to a region having poor wettability with respect to an immobilization layer forming coating solution or a solution containing a specific biopolymer, which will be described later. To do. In this embodiment, if the contact angle with water is 1 degree or more smaller than the adjacent region, the region is referred to as a hydrophilic region. Conversely, the contact angle with water is higher than the region adjacent to the region. If it is larger than 1 degree, it is determined as a water-repellent region.
[0071]
The wettability changing layer in this embodiment is a wettability changing layer in which the contact angle with water in a portion not irradiated with energy is a contact angle larger than the contact angle with water in a portion irradiated with energy by 1 degree or more. It is preferable that a wettability changing layer is used, particularly preferably 5 degrees or more, and most preferably 10 degrees or more.
[0072]
If the difference between the contact angle with water in the part not irradiated with energy and the contact angle with water in the part irradiated with energy is less than the specified range, immobilization is performed using the difference in wettability. This is because it becomes difficult to apply the layer-forming coating solution in a pattern, and it becomes difficult to fix the specific biopolymer in a pattern.
[0073]
As a specific contact angle with water in such a wettability changing layer, the contact angle with water in a portion not irradiated with energy is 30 degrees or more, particularly 60 degrees or more, and particularly 90 degrees or more. Preferably, a wettability changing layer having such a contact angle with water is preferably used. This is a portion where water repellency is required in the present embodiment where energy is not irradiated. Therefore, when the contact angle with water is small, the water repellency is not sufficient, and there is a possibility that a fixing layer forming coating liquid, which will be described later, may remain even in unnecessary parts. This is because the shape of the specific biopolymer to be converted may be disturbed and the detection accuracy of the microarray chip may be lowered.
[0074]
In addition, the contact angle with water here is measured by using a contact angle measuring device (CA-Z type manufactured by Kyowa Interface Science Co., Ltd.) with a contact angle with water (dropping a water drop from a microsyringe for 30 seconds. And later).
[0075]
In addition, when the wettability changing layer as described above is used in this embodiment, fluorine is contained in the wettability changing layer, and the fluorine content on the surface of the wettability changing layer is energy relative to the wettability changing layer. The wettability changing layer may be formed so as to be lower than that before energy irradiation due to the action of the photocatalyst contained in the photocatalyst treatment layer.
[0076]
In the microarray chip having such a feature, a pattern composed of a portion having a small fluorine content can be easily formed by pattern irradiation with energy. Here, fluorine has an extremely low surface energy. Therefore, the surface of a substance containing a large amount of fluorine has a smaller critical surface tension. Therefore, the critical surface tension of the portion having a small fluorine content is larger than the critical surface tension of the surface of the portion having a large fluorine content. This means that the portion with a low fluorine content is a hydrophilic region compared to the portion with a high fluorine content. Therefore, forming a pattern composed of a portion having a lower fluorine content than the surrounding surface forms a hydrophilic region pattern in the water-repellent region.
[0077]
Therefore, when such a wettability changing layer is used, the photocatalyst treatment layer and the wettability changing layer are arranged so as to face each other, and the pattern of the hydrophilic region is easily formed in the water-repellent region by irradiating the pattern with energy. Therefore, it is possible to easily apply an immobilization layer forming coating solution for immobilizing a specific biopolymer only to this hydrophilic region, and a microarray chip with good accuracy. can do.
[0078]
The fluorine content in the hydrophilic region having a low fluorine content formed by irradiation with energy is 10 or less, preferably 5 or less, particularly preferably 1 when the fluorine content of the portion not irradiated with energy is defined as 100. The following is preferable.
[0079]
By setting it within such a range, it is possible to make a great difference in hydrophilicity between the energy irradiated portion and the unirradiated portion. Therefore, by forming an immobilization layer on such a wettability changing layer, it becomes possible to form an immobilization layer accurately only in a hydrophilic region having a reduced fluorine content, and a microarray chip can be obtained with high accuracy. Because it can. This rate of decrease is based on weight.
[0080]
The fluorine content in such a wettability changing layer can be measured by various commonly used methods such as X-ray Photoelectron Spectroscopy, ESCA (Electron Spectroscopy for Chemical Analysis)), and any method capable of quantitatively measuring the amount of fluorine on the surface, such as X-ray fluorescence analysis and mass spectrometry.
[0081]
The material used for such a wettability changing layer is a material whose wettability changes due to the action of the photocatalyst in the photocatalyst treatment layer during the irradiation of energy, that is, the characteristics of the wettability changing layer described above. The main chain is not particularly limited as long as it has a main chain that is not easily deteriorated or decomposed by the action. For example, (1) chloro- or alkoxysilane is hydrolyzed and polycondensed by sol-gel reaction or the like to increase the strength. Examples thereof include organopolysiloxanes to be exhibited, and (2) organopolysiloxanes such as organopolysiloxanes crosslinked with reactive silicones having excellent water repellency and oil repellency.
[0082]
In the case of (1) above, the general formula:
Y_nSiX_(4-n)
(Here, Y represents an alkyl group, a fluoroalkyl group, a vinyl group, an amino group, a phenyl group or an epoxy group, X represents an alkoxyl group, an acetyl group or a halogen. N is an integer from 0 to 3. )
It is preferable that it is the organopolysiloxane which is a 1 type, or 2 or more types of hydrolysis condensate or cohydrolysis condensate of the silicon compound shown by these. Here, the number of carbon atoms of the group represented by Y is preferably in the range of 1 to 20, and the alkoxy group represented by X is a methoxy group, an ethoxy group, a propoxy group, or a butoxy group. preferable.
[0083]
In particular, an organopolysiloxane containing a fluoroalkyl group can be preferably used, and specific examples thereof include one or two or more hydrolytic condensates and cohydrolytic condensates of the following fluoroalkylsilanes. In general, those known as fluorine-based silane coupling agents can be used.
[0084]
CF₃(CF₂)₃CH₂CH₂Si (OCH₃)₃;
CF₃(CF₂)₅CH₂CH₂Si (OCH₃)₃;
CF₃(CF₂)₇CH₂CH₂Si (OCH₃)₃;
CF₃(CF₂)₉CH₂CH₂Si (OCH₃)₃;
(CF₃)₂CF (CF₂)₄CH₂CH₂Si (OCH₃)₃;
(CF₃)₂CF (CF₂)₆CH₂CH₂Si (OCH₃)₃;
(CF₃)₂CF (CF₂)₈CH₂CH₂Si (OCH₃)₃;
CF₃(C₆H₄) C₂H₄Si (OCH₃)₃;
CF₃(CF₂)₃(C₆H₄) C₂H₄Si (OCH₃)₃;
CF₃(CF₂)₅(C₆H₄) C₂H₄Si (OCH₃)₃;
CF₃(CF₂)₇(C₆H₄) C₂H₄Si (OCH₃)₃;
CF₃(CF₂)₃CH₂CH₂SiCH₃(OCH₃)₂;
CF₃(CF₂)₅CH₂CH₂SiCH₃(OCH₃)₂;
CF₃(CF₂)₇CH₂CH₂SiCH₃(OCH₃)₂;
CF₃(CF₂)₉CH₂CH₂SiCH₃(OCH₃)₂;
(CF₃)₂CF (CF₂)₄CH₂CH₂SiCH₃(OCH₃)₂;
(CF₃)₂CF (CF₂)₆CH₂CH₂Si CH₃(OCH₃)₂;
(CF₃)₂CF (CF₂)₈CH₂CH₂Si CH₃(OCH₃)₂;
CF₃(C₆H₄) C₂H₄SiCH₃(OCH₃)₂;
CF₃(CF₂)₃(C₆H₄) C₂H₄SiCH₃(OCH₃)₂;
CF₃(CF₂)₅(C₆H₄) C₂H₄SiCH₃(OCH₃)₂;
CF₃(CF₂)₇(C₆H₄) C₂H₄SiCH₃(OCH₃)₂;
CF₃(CF₂)₃CH₂CH₂Si (OCH₂CH₃)₃;
CF₃(CF₂)₅CH₂CH₂Si (OCH₂CH₃)₃;
CF₃(CF₂)₇CH₂CH₂Si (OCH₂CH₃)₃;
CF₃(CF₂)₉CH₂CH₂Si (OCH₂CH₃)₃;and
CF₃(CF₂)₇SO₂N (C₂H₅) C₂H₄CH₂Si (OCH₃)₃.
[0085]
By using a polysiloxane containing a fluoroalkyl group as described above as a binder, the water repellency of the non-energy-irradiated portion of the wettability changing layer is greatly improved. For example, the pixel portion is colored when the functional element is a color filter. The function which prevents adhesion of the functional part composition such as the ink for use is developed.
[0086]
Examples of the reactive silicone (2) include compounds having a skeleton represented by the following general formula.
[0087]
[Chemical 1]

[0088]
However, n is an integer greater than or equal to 2, R¹, R²Each represents a substituted or unsubstituted alkyl, alkenyl, aryl or cyanoalkyl group having 1 to 10 carbon atoms, and 40% or less of the total is vinyl, phenyl or phenyl halide in a molar ratio. R¹, R²Is preferably a methyl group since the surface energy is minimized, and the methyl group is preferably 60% or more by molar ratio. In addition, the chain end or side chain has at least one reactive group such as a hydroxyl group in the molecular chain.
[0089]
Moreover, you may mix the stable organosilicone compound which does not carry out a crosslinking reaction like dimethylpolysiloxane with said organopolysiloxane.
[0090]
The wettability changing layer in this embodiment can further contain a surfactant. Specifically, hydrocarbons such as NIKKOL BL, BC, BO, BB series manufactured by Nikko Chemicals Co., Ltd., ZONYL FSN, FSO manufactured by DuPont, Surflon S-141, 145 manufactured by Asahi Glass Co., Ltd., Dainippon Megafac F-141, 144 manufactured by Ink Chemical Industry Co., Ltd., Footgent F-200, F251 manufactured by Neos Co., Ltd., Unidyne DS-401, 402 manufactured by Daikin Industries, Ltd., Fluorard FC-170 manufactured by 3M Co., Ltd. Fluorine-based or silicone-based nonionic surfactants such as 176 can be used, and cationic surfactants, anionic surfactants, and amphoteric surfactants can also be used.
[0091]
In addition to the above surfactants, the wettability changing layer includes polyvinyl alcohol, unsaturated polyester, acrylic resin, polyethylene, diallyl phthalate, ethylene propylene diene monomer, epoxy resin, phenol resin, polyurethane, melamine resin, polycarbonate. , Polyvinyl chloride, polyamide, polyimide, styrene butadiene rubber, chloroprene rubber, polypropylene, polybutylene, polystyrene, polyvinyl acetate, polyester, polybutadiene, polybenzimidazole, polyacrylonitrile, epichlorohydrin, polysulfide, polyisoprene, oligomers, polymers, etc. Can be contained.
[0092]
Such a wettability changing layer can be formed by dispersing the above-described components in a solvent together with other additives as necessary to prepare a coating solution, and coating this coating solution on a substrate. it can. As the solvent to be used, alcohol-based organic solvents such as ethanol and isopropanol are preferable. The coating can be performed by a known coating method such as spin coating, spray coating, dip coating, roll coating or bead coating. In addition, in the case where an ultraviolet curable component is contained, the wettability changing layer can be formed by irradiating with ultraviolet rays and performing a curing treatment.
[0093]
In this embodiment, the thickness of the wettability changing layer is preferably 0.001 μm to 1 μm, particularly preferably in the range of 0.01 to 0.1 μm, in consideration of the change rate of wettability by the photocatalyst. is there.
[0094]
By using the wettability changing layer of the component described above in this embodiment, the action of the photocatalyst in the contacting photocatalyst processing layer is used to oxidize and decompose organic groups and additives that are part of the component. It is possible to change the wettability of the energy irradiation part to make it hydrophilic and to make a large difference in wettability with the unirradiated part. Therefore, by improving the receptivity (hydrophilicity) and repellent property (water repellency) with the fixing layer forming coating solution or the like, it is possible to obtain a microarray chip having good quality and advantageous in terms of cost.
[0095]
In addition, the wettability changing layer used in the present invention is not particularly limited as long as the wettability changing layer is changed by the action of the photocatalyst as described above. preferable. If the photocatalyst is not contained in the wettability changing layer in this way, the semi-permanent action of the photocatalyst does not reach the microarray chip when used as a microarray chip, and can be used without problems for a long period of time. That's why.
[0096]
In addition, the microarray chip substrate in this embodiment is usually formed by forming a wettability changing layer on the substrate. In this embodiment, the wettability changing layer is formed of a material having self-supporting properties. If it is, it may not contain a substrate.
[0097]
As a material for such a wettability changing layer having self-supporting property, specifically, a photocatalyst-treated layer is brought into contact with the surface and irradiated with energy to thereby apply an immobilization layer-forming coating liquid. Examples thereof include a material whose contact angle with a liquid having a surface tension equivalent to the surface tension it has changes by at least 1 ° or more, preferably 5 ° or more, particularly 10 ° or more.
[0098]
The wettability changing layer needs to be formed of a material that can transmit the irradiated energy.
[0099]
Examples of such materials include polyethylene, polycarbonate, polypropylene, polystyrene, polyester, polyvinyl fluoride, acetal resin, nylon, ABS, PTFE, methacrylic resin, phenol resin, polyvinylidene fluoride, polyoxymethylene, polyvinyl alcohol, Examples thereof include polyvinyl chloride, polyethylene terephthalate, and silicone.
[0100]
(Pattern of hydrophilic region on wettability changing layer)
The arrangement of the hydrophilic region pattern on the wettability changing layer is determined by detecting the position of the hydrophilic region (that is, the immobilization layer and the specific biopolymer formed thereon) when detecting a signal derived from the label. The position is not particularly limited as long as the position can be confirmed. For example, the position is appropriately determined according to the purpose of analysis or according to the analyzer used to detect the signal derived from the label. can do.
[0101]
Such a hydrophilic region pattern is preferably arranged in alignment in view of easy detection. Further, it is more preferable that the plurality of portions of the hydrophilic region are arranged in a line in a line at a predetermined interval, and further, such a pattern in which a plurality of such lines are arranged in parallel with each other. With such a pattern in which hydrophilic regions are arranged, it is easy to automate the process of detecting a signal derived from a label possessed by a polymer that binds to a specific biopolymer.
[0102]
The interval between the hydrophilic regions on the wettability changing layer is such that it is not affected by the adjacent hydrophilic region when detecting a signal derived from the labeling label possessed by the polymer that binds to the specific biopolymer. For example, the shortest distance between the edges of adjacent hydrophilic regions is in the range of 0.01 μm to 1 cm, preferably 0.05 μm to 5 mm. preferable. Further, the number of hydrophilic regions on the wettability changing layer is not particularly limited, and depends on the size of the base material. For example, several to tens of thousands (generally several tens to one to one to 20,000).
[0103]
In the present invention, such a wettability changing layer pattern forming method is such that, in the present invention, a photocatalyst-treated layer containing a photocatalyst is disposed with the wettability changing layer facing each other, and an energy pattern irradiation using a photomask Can be performed. Further, as will be described later, when the photocatalyst treatment layer side light-shielding portion is formed on the photocatalyst treatment layer side substrate, the entire energy may be irradiated.
[0104]
(2) Base material
Next, the substrate will be described. In this embodiment, when the above-described wettability changing layer is poor in self-supporting property, it may be a microarray chip base material in which the wettability changing layer is formed on the base material.
[0105]
Materials used for such substrates include organic or inorganic materials that are substantially insoluble in water or organic solvents commonly used in nucleic acid hybridization assays or immunological assays, such as silicon, Glass, magnetic metal or nonmagnetic metal, plastic, etc. can be mentioned. In particular, silicon, various plastics (for example, polycarbonate, polystyrene, polypropylene, or the like), quartz, glass, or the like is preferably used. The shape of such a substrate is not particularly limited as long as it has at least one flat surface, but it can be said that it is preferably a plate shape or a film shape.
[0106]
B. Pattern formation process
Next, the pattern formation process in this aspect is demonstrated. This step uses a photocatalyst treatment layer side substrate in which a photocatalyst treatment layer containing at least a photocatalyst is formed on the substrate, and the microarray chip substrate and the photocatalyst treatment layer side substrate are combined with the photocatalyst treatment layer and the wetting layer. In the process of forming a pattern due to the difference in wettability on the surface of the wettability changing layer by irradiating energy from a predetermined direction after the property changing layers are arranged facing each other and with a gap of 200 μm or less. is there. Hereinafter, the photocatalyst processing layer side substrate and energy irradiation used in this step will be described.
[0107]
In addition, this process is a process performed also in the 2nd embodiment mentioned later besides this aspect. Therefore, in the following description of this process, in order to correspond to both aspects of the present invention, the above-described wettability changing layer and a decomposition / removal layer described later may be collectively referred to as a characteristic changing layer.
[0108]
(1) Photocatalyst processing layer side substrate
This step is characterized in that a photocatalyst processing layer side substrate is used in energy irradiation described later. This photocatalyst processing layer side substrate has at least a photocatalyst processing layer and a substrate, and is usually formed by forming a thin film photocatalyst processing layer formed by a predetermined method on the substrate. Moreover, the thing in which the photocatalyst processing layer side light-shielding part formed in pattern shape was formed can also be used for this photocatalyst processing layer side board | substrate.
[0109]
a. Photocatalyst treatment layer
The photocatalyst treatment layer used in the present invention is not particularly limited as long as the photocatalyst in the photocatalyst treatment layer changes the characteristics of the characteristic change layer, and is composed of a photocatalyst and a binder. It may be a thing, and the thing formed into a film by the photocatalyst simple substance may be used. Further, the wettability of the surface may be particularly hydrophilic or water repellent.
[0110]
The photocatalyst treatment layer used in the present invention may be formed on the entire surface of the substrate 1 as shown in FIG. 1A, for example, but for example as shown in FIG. The photocatalyst processing layer 2 may be formed on the pattern.
[0111]
By forming the photocatalyst treatment layer in a pattern in this way, as will be described in the energy irradiation described later, when the photocatalyst treatment layer is disposed with a predetermined distance from the characteristic change layer and irradiated with energy, It is not necessary to perform pattern irradiation using a mask or the like, and by irradiating the entire surface, a pattern with changed characteristics can be formed on the characteristic change layer.
[0112]
The patterning method of the photocatalyst processing layer is not particularly limited, but can be performed by, for example, a photolithography method.
[0113]
In addition, since the characteristics of only the part on the characteristic change layer facing the photocatalyst treatment layer actually change, the energy irradiation direction is such that the part where the photocatalyst treatment layer and the characteristic change layer face is irradiated with energy. As long as it irradiates, it may irradiate from what direction, and also has the advantage that irradiation energy is not limited to parallel things, such as parallel light.
[0114]
Thus, the action mechanism of the photocatalyst represented by titanium dioxide as described later in the photocatalyst treatment layer is not necessarily clear, but the carrier generated by light irradiation reacts directly with a nearby compound, or It is considered that the active oxygen species generated in the presence of oxygen and water change the chemical structure of organic matter. In the present invention, it is considered that this carrier acts on the compound in the property change layer disposed in the vicinity of the photocatalyst treatment layer.
[0115]
As the photocatalyst used in the present invention, for example, titanium dioxide (TiO 2) known as an optical semiconductor.₂), Zinc oxide (ZnO), tin oxide (SnO)₂), Strontium titanate (SrTiO)₃), Tungsten oxide (WO₃), Bismuth oxide (Bi₂O₃), And iron oxide (Fe₂O₃1) or a mixture of two or more selected from these.
[0116]
In the present invention, titanium dioxide is particularly preferably used because it has a high band gap energy, is chemically stable, has no toxicity, and is easily available. Titanium dioxide includes an anatase type and a rutile type, and both can be used in the present invention, but anatase type titanium dioxide is preferred. Anatase type titanium dioxide has an excitation wavelength of 380 nm or less.
[0117]
Examples of such anatase type titanium dioxide include hydrochloric acid peptizer type anatase type titania sol (STS-02 manufactured by Ishihara Sangyo Co., Ltd. (average particle size 7 nm), ST-K01 manufactured by Ishihara Sangyo Co., Ltd.), nitric acid solution An anatase type titania sol (TA-15 manufactured by Nissan Chemical Co., Ltd. (average particle size 12 nm)) and the like can be mentioned.
[0118]
The smaller the particle size of the photocatalyst, the more effective the photocatalytic reaction occurs. The average particle size is preferably 50 nm or less, and it is particularly preferable to use a photocatalyst of 20 nm or less.
[0119]
The photocatalyst treatment layer in the present invention may be formed by the photocatalyst alone as described above, or may be formed by mixing with a binder.
[0120]
In the case of a photocatalyst treatment layer composed only of a photocatalyst, the efficiency with respect to the change of the characteristics on the characteristic change layer is improved, which is advantageous in terms of cost such as shortening of the treatment time. On the other hand, in the case of a photocatalyst treatment layer comprising a photocatalyst and a binder, there is an advantage that the formation of the photocatalyst treatment layer is easy.
[0121]
Examples of a method for forming a photocatalyst treatment layer composed of only a photocatalyst include a method using a vacuum film forming method such as a sputtering method, a CVD method, or a vacuum deposition method. By forming the photocatalyst treatment layer by a vacuum film formation method, it is possible to obtain a photocatalyst treatment layer that is a uniform film and contains only the photocatalyst, and this can uniformly change the characteristics on the characteristic change layer. Since it is possible and consists only of a photocatalyst, it is possible to change the characteristics on the characteristic change layer more efficiently than in the case of using a binder.
[0122]
Examples of a method for forming a photocatalyst treatment layer composed only of a photocatalyst include a method in which amorphous titania is formed on a substrate when the photocatalyst is titanium dioxide, and then the phase is changed to crystalline titania by firing. As the amorphous titania used here, for example, hydrolysis, dehydration condensation, tetraethoxytitanium, tetraisopropoxytitanium, tetra-n-propoxytitanium, tetrabutoxytitanium, titanium inorganic salts such as titanium tetrachloride and titanium sulfate, An organic titanium compound such as tetramethoxytitanium can be obtained by hydrolysis and dehydration condensation in the presence of an acid. Next, it can be modified to anatase titania by baking at 400 ° C. to 500 ° C. and modified to rutile type titania by baking at 600 ° C. to 700 ° C.
[0123]
Moreover, when using a binder, what has the high bond energy that the main frame | skeleton of a binder is not decomposed | disassembled by photoexcitation of said photocatalyst is preferable, for example, organopolysiloxane etc. can be mentioned.
[0124]
When organopolysiloxane is used as a binder in this way, the photocatalyst treatment layer prepares a coating liquid by dispersing the photocatalyst and the organopolysiloxane as a binder in a solvent together with other additives as necessary. It can be formed by applying this coating solution on a substrate. As the solvent to be used, alcohol-based organic solvents such as ethanol and isopropanol are preferable. The coating can be performed by a known coating method such as spin coating, spray coating, dip coating, roll coating or bead coating. When the binder contains an ultraviolet curable component, the photocatalyst treatment layer can be formed by irradiating ultraviolet rays to carry out the curing treatment.
[0125]
An amorphous silica precursor can be used as the binder. This amorphous silica precursor has the general formula SiX₄X is preferably a silicon compound such as halogen, methoxy group, ethoxy group, or acetyl group, silanol as a hydrolyzate thereof, or polysiloxane having an average molecular weight of 3000 or less.
[0126]
Specific examples include tetraethoxysilane, tetraisopropoxysilane, tetra-n-propoxysilane, tetrabutoxysilane, and tetramethoxysilane. In this case, the amorphous silica precursor and the photocatalyst particles are uniformly dispersed in a non-aqueous solvent, hydrolyzed with moisture in the air on the substrate to form silanol, and then at room temperature. A photocatalyst treatment layer can be formed by dehydration condensation polymerization. If dehydration condensation polymerization of silanol is carried out at 100 ° C. or higher, the degree of polymerization of silanol increases and the strength of the film surface can be improved. Moreover, these binders can be used individually or in mixture of 2 or more types.
[0127]
When the binder is used, the content of the photocatalyst in the photocatalyst treatment layer can be set in the range of 5 to 60% by weight, preferably 20 to 40% by weight. The thickness of the photocatalyst treatment layer is preferably in the range of 0.05 to 10 μm.
[0128]
In addition to the photocatalyst and the binder, the photocatalyst treatment layer can contain a surfactant and an additive. Since these surfactants and additives are the same as those in the “(1) wettability changing layer” described above, description thereof is omitted here.
[0129]
b. substrate
In the present invention, as shown in FIG. 1, the photocatalyst processing layer side substrate 3 has at least a substrate 1 and a photocatalyst processing layer 2 formed on the substrate 1.
[0130]
At this time, the material constituting the substrate to be used is appropriately selected depending on the direction of energy irradiation to be described later.
[0131]
That is, for example, in the case where an opaque base material for microarray chips is used as the base material, the energy irradiation direction is necessarily from the photocatalyst processing layer side substrate side, and as shown in FIG. Must be disposed on the photocatalyst processing layer side substrate 3 side and irradiated with energy. Further, as will be described later, even when a photocatalyst treatment layer side light-shielding portion is formed in a predetermined pattern in advance on the photocatalyst treatment layer side substrate and the pattern is formed using this photocatalyst treatment layer side light-shielding portion, It is necessary to irradiate energy from the side substrate side. In such a case, it is necessary that the substrate has transparency.
[0132]
On the other hand, if the microarray chip substrate is transparent, it is possible to irradiate energy by arranging a photomask on the microarray chip substrate side. In such a case, the substrate is not particularly required to be transparent.
[0133]
The substrate used in the present invention may be a flexible substrate such as a resin film, or may be a non-flexible substrate such as a glass substrate. This is appropriately selected depending on the energy irradiation method described later.
[0134]
Thus, the substrate used for the photocatalyst processing layer side substrate in the present invention is not particularly limited in material, but in the present invention, the photocatalyst processing layer side substrate is used repeatedly. A material having a predetermined strength and having a surface having good adhesion to the photocatalyst treatment layer is preferably used.
[0135]
Specific examples include glass, ceramic, metal, and plastic.
[0136]
Note that an anchor layer may be formed on the substrate in order to improve the adhesion between the substrate surface and the photocatalyst treatment layer. Examples of such an anchor layer include silane-based and titanium-based coupling agents.
[0137]
c. Photocatalyst treatment layer side shading part
As the photocatalyst processing layer side substrate used in the present invention, a substrate on which a photocatalyst processing layer side light-shielding portion formed in a pattern is formed may be used. By using the photocatalyst processing layer side substrate having the photocatalyst processing layer side light-shielding portion in this way, it is not necessary to use a photomask or perform drawing irradiation with laser light when irradiating energy. Therefore, since alignment between the photocatalyst processing layer side substrate and the photomask is unnecessary, it is possible to make a simple process, and an expensive apparatus necessary for drawing irradiation is unnecessary, so that the cost is reduced. Has the advantage of being advantageous.
[0138]
The photocatalyst processing layer side substrate having such a photocatalyst processing layer side light shielding part can be set to the following two embodiments depending on the formation position of the photocatalyst processing layer side light shielding part.
[0139]
For example, as shown in FIG. 9, a photocatalyst processing layer side light shielding portion 31 is formed on the substrate 1, and a photocatalyst processing layer 2 is formed on the photocatalyst processing layer side light shielding portion 31 to form a photocatalyst processing layer side substrate. 3 is an embodiment. For example, as shown in FIG. 10, the photocatalyst processing layer 2 is formed on the substrate 1, and the photocatalyst processing layer side light-shielding portion 31 is formed thereon to form the photocatalyst processing layer side substrate 3. is there.
[0140]
In any embodiment, compared to the case of using a photomask, the photocatalyst processing layer side light shielding portion is disposed in the vicinity of the portion where the photocatalyst processing layer and the characteristic change layer are located with a gap. Since the influence of energy scattering in the substrate or the like can be reduced, energy pattern irradiation can be performed very accurately.
[0141]
Furthermore, in the embodiment in which the photocatalyst treatment layer side light shielding portion is formed on the photocatalyst treatment layer, when the photocatalyst treatment layer and the characteristic change layer are arranged with a predetermined gap, By making the film thickness coincide with the width of the gap, there is an advantage that the photocatalyst processing layer side light-shielding portion can be used as a spacer for making the gap constant.
[0142]
That is, when the photocatalyst treatment layer and the characteristic change layer are arranged in contact with each other with a predetermined gap, the photocatalyst treatment layer side light shielding portion and the characteristic change layer are arranged in close contact with each other. The predetermined gap can be made accurate, and in this state, by irradiating energy from the photocatalyst processing layer side substrate, a pattern can be accurately formed on the characteristic change layer.
[0143]
The formation method of such a photocatalyst processing layer side light shielding part is not particularly limited, and is appropriately selected according to the characteristics of the formation surface of the photocatalyst processing layer side light shielding part, the shielding property against the required energy, and the like. Used.
[0144]
For example, it may be formed by forming a metal thin film of chromium or the like having a thickness of about 1000 to 2000 mm by a sputtering method, a vacuum deposition method, or the like, and patterning the thin film. As this patterning method, a normal patterning method such as sputtering can be used.
[0145]
Alternatively, a method may be used in which a layer containing light-shielding particles such as carbon fine particles, metal oxides, inorganic pigments, and organic pigments in a resin binder is formed in a pattern. As the resin binder to be used, polyimide resin, acrylic resin, epoxy resin, polyacrylamide, polyvinyl alcohol, gelatin, casein, cellulose, or a mixture of one or more kinds, photosensitive resin, or O / A W emulsion type resin composition, for example, an emulsion of a reactive silicone can be used. The thickness of such a resin photocatalyst treatment layer side light-shielding part can be set within a range of 0.5 to 10 μm. As a method for patterning the light shielding part on the resin photocatalyst treatment layer side, a generally used method such as a photolithography method or a printing method can be used.
[0146]
In the above description, the two positions of the photocatalyst treatment layer side light shielding portion are described between the substrate and the photocatalyst treatment layer and the surface of the photocatalyst treatment layer. In addition, the photocatalyst treatment layer of the substrate is formed. It is also possible to adopt a mode in which the photocatalyst processing layer side light shielding portion is formed on the surface that is not provided. In this aspect, for example, a case where a photomask is attached to the surface to such an extent that it can be attached and detached can be considered.
[0147]
d. Primer layer
In the present invention, when the photocatalyst treatment layer side light-shielding portion is formed in a pattern on the substrate as described above and the photocatalyst treatment layer is formed thereon to form the photocatalyst treatment layer side substrate, the photocatalyst treatment layer It is preferable to form a primer layer between the side light-shielding part and the photocatalyst treatment layer.
[0148]
The action and function of this primer layer are not necessarily clear, but by forming a primer layer between the photocatalyst treatment layer-side light-shielding part and the photocatalyst treatment layer, the primer layer has characteristics of the property change layer due to the action of the photocatalyst. Impurities from the openings existing between the photocatalyst processing layer side light shielding part and the photocatalyst processing layer side light shielding part, which are factors that hinder the change, in particular, residues generated when patterning the photocatalyst processing layer side light shielding part, metal, metal It is considered that it has a function of preventing diffusion of impurities such as ions. Therefore, by forming the primer layer, the characteristic change process proceeds with high sensitivity, and as a result, a high-resolution pattern can be obtained.
[0149]
In the present invention, the primer layer prevents the impurities present in the opening formed between the photocatalyst processing layer side light shielding part as well as the photocatalyst processing layer side light shielding part from affecting the action of the photocatalyst. The primer layer is preferably formed over the entire light-shielding portion on the photocatalyst processing layer side including the opening.
[0150]
FIG. 11 shows an example of the photocatalyst processing layer side substrate on which such a primer layer is formed. A primer layer 32 is formed on the surface of the substrate 1 on which the photocatalyst processing layer side light shielding portion 31 is formed, on the side where the photocatalyst processing layer side light shielding portion 31 is formed, and the photocatalyst processing layer 2 is formed on the surface of the primer layer 32. Is formed.
[0151]
The configuration in which the photocatalyst processing layer side light shielding portion is formed in a pattern on the substrate is a general photomask configuration. Therefore, it can be said that this primer layer is a photocatalyst treatment layer formed on the photomask via the primer layer.
[0152]
The primer layer in the present invention is not particularly limited as long as the photocatalyst treatment layer and the photomask are arranged so as not to be in physical contact. That is, the primer layer only needs to be formed so that the photocatalyst processing layer side light-shielding portion of the photomask does not contact the photocatalyst processing layer.
[0153]
The material constituting the primer layer is not particularly limited, but an inorganic material that is not easily decomposed by the action of the photocatalyst is preferable. Specific examples include amorphous silica. When such amorphous silica is used, the precursor of amorphous silica is represented by the general formula SiX.₄X is a silicon compound such as halogen, methoxy group, ethoxy group, or acetyl group, and a hydrolyzate thereof, silanol, or polysiloxane having an average molecular weight of 3000 or less is preferable.
[0154]
The thickness of the primer layer is preferably in the range of 0.001 μm to 1 μm, particularly preferably in the range of 0.001 μm to 0.1 μm.
[0155]
(2) Energy irradiation
In the present invention, the photocatalyst treatment layer side substrate and the microarray chip base material are disposed with the photocatalyst treatment layer and the characteristic change layer facing each other with a gap of 200 μm or less, and then irradiated with energy from a predetermined direction. A pattern forming step is performed. The gap of 200 μm or less includes a state where the photocatalyst processing layer and the characteristic change layer are in contact with each other.
[0156]
Thus, by disposing the photocatalyst treatment layer and the surface of the characteristic change layer at a predetermined interval, oxygen, water, and active oxygen species generated by the photocatalytic action are easily desorbed. In other words, it is not preferable to arrange them at a distance from the above range, because the generated active oxygen species are difficult to reach the characteristic change layer, and the characteristic change rate may be reduced. On the other hand, if the distance between the photocatalyst treatment layer and the characteristic change layer is too narrow, desorption of oxygen, water, and active oxygen species generated by the photocatalytic action becomes difficult, resulting in a slow characteristic change rate. This is not preferable because of the possibility.
[0157]
In the present invention, the gap has a very good pattern accuracy, a high sensitivity of the photocatalyst, and therefore a good characteristic change efficiency, so that it is particularly within the range of 0.2 μm to 10 μm, preferably 1 μm to 5 μm. It is preferable to be within the range.
[0158]
On the other hand, when processing is performed on a microarray chip substrate having a large area of, for example, 300 mm × 300 mm, the photocatalyst processing layer side substrate and the microarray chip substrate are not contacted and the fine gap as described above is formed between the substrate and the microarray chip substrate. It is extremely difficult to provide between the two. Therefore, when the microarray chip substrate has a relatively large area, the gap is preferably in the range of 10 to 100 μm, particularly in the range of 50 to 75 μm. By setting the gap within such a range, there is no problem of pattern accuracy deterioration such as blurring of the pattern or problems such as deterioration of photocatalyst sensitivity and deterioration of efficiency of characteristic change. This is because there is an effect that unevenness of the characteristic change on the layer does not occur.
[0159]
When energy is applied to the microarray chip substrate having a relatively large area as described above, the gap setting in the positioning device between the photocatalyst processing layer side substrate and the microarray chip substrate in the energy irradiation apparatus is set to 10 μm to 200 μm. It is preferable to set within a range of 25 μm to 75 μm. By setting the set value within such a range, the pattern accuracy is not significantly lowered and the photocatalyst sensitivity is not greatly deteriorated, and the photocatalyst processing layer side substrate and the microarray chip base material are not in contact with each other. This is because they can be arranged.
[0160]
In the present invention, such an arrangement state with a gap need only be maintained at least during energy irradiation.
[0161]
As a method of arranging such a very narrow gap uniformly and arranging the photocatalyst processing layer and the characteristic change layer, for example, a method using a spacer can be cited. By using the spacer in this way, a uniform gap can be formed, and the portion in contact with the spacer is not affected by the photocatalyst action on the surface of the characteristic change layer. It is possible to form a predetermined pattern on the characteristic change layer by having the same pattern as.
[0162]
In the present invention, such a spacer may be formed as one member. However, for simplification of the process, as described in the column of the photocatalyst treatment layer side substrate, the photocatalyst of the photocatalyst treatment layer side substrate is used. It is preferable to form on the treatment layer surface. In the description of the photocatalyst processing layer side substrate, the photocatalyst processing layer side light shielding portion has been described. However, in the present invention, such a spacer protects the surface so that the photocatalytic action does not reach the surface of the property change layer. Therefore, it may be formed of a material that does not have a function of shielding the irradiated energy.
[0163]
Next, in a state where the contact state as described above is maintained, energy irradiation is performed on the contacted portion. Usually, the wavelength of light used for such energy irradiation is set in the range of 400 nm or less, preferably in the range of 380 nm or less. This is because, as described above, the preferred photocatalyst used in the photocatalyst treatment layer is titanium dioxide, and the light having the above-described wavelength is preferable as the energy for activating the photocatalytic action by the titanium dioxide.
[0164]
Examples of light sources that can be used for such energy irradiation include mercury lamps, metal halide lamps, xenon lamps, excimer lamps, and various other light sources.
[0165]
In addition to the method of performing pattern irradiation such as through a photomask using a light source as described above, it is also possible to use a method of drawing and irradiating in a pattern using a laser such as excimer or YAG.
[0166]
In addition, the energy irradiation amount at the time of energy irradiation is an irradiation amount necessary for the surface of the characteristic change layer to change the characteristic of the surface of the characteristic change layer by the action of the photocatalyst in the photocatalyst treatment layer.
[0167]
At this time, it is preferable in that the sensitivity can be increased by irradiating the photocatalyst treatment layer with energy while heating and the characteristic can be changed efficiently. Specifically, it is preferable to heat within a range of 30 ° C to 80 ° C.
[0168]
The energy irradiation direction in the present invention is determined by whether or not the photocatalyst treatment layer side light-shielding portion is formed on the photocatalyst treatment layer side substrate and whether or not the photocatalyst treatment layer side substrate or the microarray chip substrate is transparent. The
[0169]
That is, when the photocatalyst processing layer side light shielding part is formed on the photocatalyst processing layer side substrate, it is necessary to irradiate energy from the photocatalyst processing layer side substrate side, and in this case, the photocatalyst processing layer side substrate is irradiated. It needs to be transparent to energy. In this case, when the photocatalyst treatment layer side light shielding portion is formed on the photocatalyst treatment layer and this photocatalyst treatment layer side light shielding portion is used so as to function as a spacer as described above, the irradiation direction is It may be from the photocatalyst processing layer side substrate side or from the microarray chip substrate side.
[0170]
Further, as described above, the irradiation direction when the photocatalyst processing layer is formed in a pattern is irradiated from any direction as long as the energy is applied to the portion where the photocatalyst processing layer and the property change layer are in contact with each other. May be.
[0171]
Similarly, in the case of using the above-described spacer, irradiation may be performed from any direction as long as energy is applied to the contact portion.
[0172]
In the case of using a photomask, energy is irradiated from the side where the photomask is arranged. In this case, either the substrate or base material on which the photomask is disposed, that is, either the photocatalyst processing layer side substrate or the microarray chip base material needs to be transparent.
[0173]
When the energy irradiation as described above is completed, the photocatalyst processing layer side substrate is moved away from the contact position with the property change layer, and thereby, for example, the hydrophilic region 8 and the wettability changed as shown in FIG. A pattern composed of the water repellent region 9 is formed on the wettability changing layer 5. On the other hand, in the case of the decomposition removal layer in the second embodiment described later, as shown in FIG. 2C, the decomposition removal layer 21 formed in a pattern is obtained.
[0174]
C. Removal process
In this embodiment, after the energy irradiation, the photocatalyst processing layer substrate is removed from the microarray chip substrate. As a result, a wettability changing layer having a pattern formed of a hydrophilic region and a water repellent region on the surface is obtained.
[0175]
D. Immobilization layer formation process
Next, the fixing layer forming step in this embodiment will be described. This step is a step of forming an immobilization layer on the hydrophilic region according to the pattern of the wettability changing layer composed of the hydrophilic region and the water repellent region.
[0176]
Hereinafter, the immobilization layer formed into a pattern by this step will be described.
[0177]
(Fixed layer)
In this embodiment, an immobilization layer for immobilizing a specific biopolymer described later is formed on the hydrophilic region formed in a pattern on the wettability changing layer.
[0178]
As the immobilizing agent used in the immobilizing layer, various agents can be used according to the immobilizing means for the specific biopolymer.
[0179]
As such an immobilization means, a binding method known to those skilled in the art can be used. For example, an immobilization method by electrostatic coupling, an immobilization method by covalent bonding, or the like can be used.
[0180]
In the case of the above-mentioned immobilization means by electrostatic binding, a solution in which a substance having a charge opposite to that of the specific biopolymer is dissolved as a fixing agent is prepared as an immobilization layer-forming coating solution. The immobilization layer is formed by applying the hydrophilic region on the wettability changing layer formed in a pattern.
[0181]
For example, when the specific biopolymer is an oligonucleotide, since the oligonucleotide has an anionic property, a coating solution using a substance having a cationic property as a fixing agent is used. The immobilization layer is formed by applying this and drying.
[0182]
In this case, examples of the cationic immobilizing agent to be used include poly L-lysine, poly (allylamine hydrochloride), poly (ethyleneimine), poly (diallyldimethylammonium chloride) and the like. L-lysine is preferred.
[0183]
On the other hand, as an immobilization method using a covalent bond (chemical bond), a compound having a functional group (for example, a carboxyl group, an amino group, or a hydroxyl group) is used as an immobilizing agent for an immobilizing agent-forming coating material. A method of forming and binding a specific biopolymer to this can be mentioned. Examples of such a fixing agent include compounds having a carboxyl group or a primary amino group as a functional group.
[0184]
Specifically, a silane coupling agent containing an amino group is attached to the hydrophilic region formed in a pattern on the wettability changing layer. This uses a solution containing the above silane coupling agent, and is applied to the entire surface by a dip coating method, a transfer method, a spin coating method, etc., and a silane coupling agent is attached only to the hydrophilic region, an inkjet, a dispenser, etc. It is possible to use a method of applying only the hydrophilic region. The silane coupling agent containing a coated amino group spreads in the hydrophilic region and does not remain in the water-repellent region. After coating, the hydrophilic region and the immobilizing agent can be bonded by heating as necessary. A microarray chip can be formed by binding a specific biopolymer on the immobilization layer thus formed.
[0185]
Strongly immobilized on the membrane by covalently bonding the amino group of the silane coupling agent, which is the immobilizing agent, to a biopolymer whose end is modified with an amino group, specifically, the terminal amino group of an oligonucleotide or the like can do.
[0186]
In this case, as the silane coupling agent having an amino group to be used, N-β (aminoethyl) γ-aminopropyltrimethoxysilane, N-β (aminoethyl) γ-aminopropylmethyldimethoxysilane, γ-aminopropyl is used. Examples include triethoxysilane, N-phenyl-γ-aminopropyltrimethoxysilane, and the like.
[0187]
In addition, as another immobilization method by covalent bond between the immobilization layer and the specific biopolymer, for example, when the biopolymer is an oligonucleotide, a method of covalently bonding the 5 ′ end to the functional group It has been known. Specifically, first, the 5 ′ end of the oligonucleotide is reacted with a maleimide compound to introduce a maleimide group at the 5 ′ end of the oligonucleotide. Suitable maleimide compounds include sulfosuccinimidyl-4- (N-maleimidomethyl) cyclohexane-1-carboxylate and the like. Separately from the above reaction, the immobilization agent having an amino group as a functional group and succinimidyl-S-acetylthioacetate are reacted, and then deacetylation is performed using hydroxyramine, whereby the immobilization agent is converted into SH. Giving a group. By reacting the SH group on the immobilizing agent thus imparted with the maleimide group introduced at the 5 ′ end of the oligonucleotide prepared by introducing a maleimide group at the 5 ′ end, the oligonucleotide and the immobilizing agent are reacted. The specific biopolymer can be immobilized on the immobilization layer.
[0188]
In addition to the methods described above, many immobilization methods such as a method using a biotin-avidin system can also be used.
[0189]
As will be described later, the immobilization layer used in this embodiment preferably does not spread from the immobilization layer when a liquid containing a specific biopolymer is attached thereon. Therefore, there is provided an immobilizing agent in which the surface wettability of the immobilization layer is smaller than the surrounding wettability changing layer region (water repellent region) by 1 degree or more, preferably 5 degrees or more in contact angle with water. It is preferably selected and used.
[0190]
E. Specific biopolymer immobilization process
Finally, the specific biopolymer immobilization step in this embodiment will be described. This step is a step of attaching a specific biopolymer onto the immobilization layer formed in a pattern by the immobilization layer forming step.
[0191]
Hereinafter, the specific biopolymer formed in this step will be described.
[0192]
(Specific biopolymer)
In the microarray chip of this embodiment, a specific biopolymer is immobilized on the immobilization layer. Such a specific biopolymer is not particularly limited as long as it is a biopolymer capable of specifically binding to a predetermined biopolymer.
[0193]
Such specific binding includes, for example, a combination of a nucleic acid (eg, oligonucleotide or polynucleotide) and a complementary nucleic acid, a combination of an antigen and an antibody (or antibody fragment), a receptor and its Combinations with ligands (eg hormones, cytokines, neurotransmitters or lectins), combinations of enzymes with their ligands (eg enzyme substrate analogs, coenzymes, modulators or inhibitors), enzyme analogs and their enzymes The combination with the substrate of the enzyme used as the base of the analog, or the combination of lectin and sugar can be mentioned. The “enzyme analog” means a substance having a high specific affinity with a substrate for the original enzyme but showing no catalytic activity. In addition, each of the compounds in each of the above combinations can be a “specific biopolymer” and the other can be a substance to be detected. For example, in the “combination of antigen and antibody”, when the antigen is “a substance to be detected”, the antibody can be “specific biopolymer”, and conversely, the antibody is “detected” In the case of “substance”, the antigen can be “specific biopolymer”.
[0194]
For example, when the microarray chip of this embodiment is applied to a nucleic acid hybridization assay, it binds complementarily to the nucleic acid (eg, oligonucleotide or polynucleotide) that is the substance to be detected as the specific biopolymer. Oligonucleotides or polynucleotides that can be used. As used herein, “oligonucleotide” or “polynucleotide” includes 2′-deoxyribonucleic acid (DNA), ribonucleic acid (RNA), and peptide nucleic acid (PNA). PNA is an artificial nucleic acid obtained by converting a DNA phosphodiester bond into a peptide bond. The chain length of the oligonucleotide or polynucleotide bound to the insoluble particle can be appropriately selected depending on the purpose of analysis, and is based on, for example, the chain length of the complementary sequence of DNA, RNA, or PNA to be captured. Can be determined.
[0195]
Synthesis of the oligonucleotide used as the specific biopolymer can be generally performed using an automatic synthesizer. In addition, if necessary, an oligonucleotide having a modified terminal amino group or other group can be produced using an automatic synthesizer. Alternatively, an unmodified oligonucleotide can be synthesized in advance using an automatic synthesizer, and the above-mentioned unmodified oligonucleotide can be modified as necessary. Methods for modifying oligonucleotides are well known, and for example, amines and fluorescein can be added. Oligonucleotides modified in this way have a greater affinity for the immobilization layer than unmodified oligonucleotides. In addition, synthesis of a polynucleotide (for example, cDNA) used as a specific biopolymer can be performed using a normal genetic engineering technique, for example, a PCR (polymerase chain reaction) method.
[0196]
When the microarray chip of this embodiment is applied to an immunological assay, an antigen (including a hapten) or an antibody that specifically binds to a substance to be detected can be used as the specific biopolymer. . In this case, the substance to be detected is not particularly limited as long as it is a component generally contained in a test sample and can be detected immunologically. For example, various proteins, polysaccharides, lipids, fungal bodies, various environmental substances, and the like can be given. More specifically, immunoglobulins (eg, IgG, IgM, or IgA), infection-related markers (eg, HBs antigen, HBs antibody, HIV-1 antibody, HIV-2 antibody, HTLV-1 antibody, or treponema antibody) Tumor associated antigens (eg, AFP, CRP, or CEA), coagulation fibrinolytic markers (eg, plasminogen, antithrombin-III, D-dimer, TAT, or PPI), antiepileptic drugs (eg, hormones), Examples include various drugs (for example, digoxin), bacterial cells (for example, O-157 or Salmonella) or their endotoxins or exotoxins, microorganisms, enzymes, residual agricultural chemicals, environmental hormones, and the like. As the antibody used as the specific biopolymer, either a polyclonal antibody or a monoclonal antibody obtained by a well-known method can be used. In addition, the antibody may be a protein [eg, enzyme (eg, pepsin or papain)] [eg, antibody fragment (eg, Fab, Fab ′, F (ab ′)₂Or Fv)] can also be used.
[0197]
2. Second embodiment
Next, a method for manufacturing a microarray chip in the second embodiment of the present invention will be described.
[0198]
In the second embodiment, the immobilization layer is patterned using a decomposition / removal layer that is decomposed and removed by the action of a photocatalyst accompanying energy irradiation. Furthermore, in this aspect, it can be divided into the aspect which forms a fixed layer using the difference in wettability of a decomposition removal layer and a base material, and the aspect which uses the decomposition removal layer itself as a fixed layer. Hereinafter, the description will be made separately for each aspect.
[0199]
A. When the immobilization layer is formed using the difference in wettability between the decomposition removal layer and the substrate
In such a microarray chip manufacturing method, a decomposition removal layer that is decomposed and removed by the action of a photocatalyst accompanying energy irradiation and has a contact angle with a liquid different from that of the substrate is formed on the substrate. Using the microarray chip base material adjusting step for adjusting the base material and the photocatalyst processing layer side substrate on which the photocatalyst processing layer containing at least the photocatalyst is formed on the substrate, the microarray chip base material and the photocatalyst processing layer side After disposing the substrate so that the photocatalyst treatment layer and the decomposition / removal layer face each other with a gap of 200 μm or less, the decomposition / removal layer is formed in a pattern by irradiating energy from a predetermined direction. Pattern forming step, and removing the photocatalyst processing layer side substrate from the microarray chip substrate In accordance with the pattern of the decomposition / removal layer, an immobilization layer forming step for forming an immobilization layer on the decomposition / removal layer or the substrate, and a specific for immobilizing a specific biopolymer on the immobilization layer And a biopolymer fixing step.
[0200]
The manufacturing method of the microarray chip in this embodiment will be described with reference to the drawings.
[0201]
FIG. 2 shows an example of a manufacturing method of the microarray chip of this embodiment. First, a photocatalyst treatment layer side substrate 3 in which a photocatalyst treatment layer 2 made of at least a photocatalyst is formed on a substrate 1 is prepared. Furthermore, apart from the photocatalyst processing layer side substrate 3, a base material 4 is prepared, and the base material 4 is decomposed and removed by the action of a photocatalyst accompanying energy irradiation, and has a wettability different from that of the base material 4. The removal layer 21 is formed, and the microarray chip substrate 6 is prepared (see FIG. 2A).
[0202]
Further, after the photocatalyst processing layer side substrate 3 and the microarray chip substrate 6 are arranged with the photocatalyst processing layer 2 and the decomposition removal layer 21 facing each other and with a predetermined gap, the energy is passed through the photomask 7. Is irradiated in a pattern (see FIG. 2B).
[0203]
Next, when the photocatalyst processing layer side substrate 3 is removed from the microarray chip substrate 6, the microarray chip substrate 6 having the decomposition removal layer 21 formed in a pattern on the substrate 4 is obtained (FIG. 2C). reference).
[0204]
Moreover, since the wettability is different between the decomposition removal layer 21 and the substrate 4, the pattern due to the difference in wettability was formed on the substrate 4 by forming the decomposition removal layer 21 in a pattern. become. By utilizing this difference in wettability, the fixing layer 10 is formed on the base material 4 where the decomposition removal layer 21 is decomposed and removed (see FIG. 2D). FIG. 2 shows a case where the fixing layer 10 is formed on the base material 4 where the decomposition / removal layer 21 is decomposed and removed, but the present invention is not limited to such a case. In the case where the substrate has a larger contact angle with respect to the liquid, an immobilization layer is formed on the decomposition removal layer.
[0205]
Finally, a microarray chip can be manufactured by attaching a specific biopolymer 11 on the immobilization layer 10 (see FIG. 2 (e)).
[0206]
According to this embodiment, no post-treatment is required after energy irradiation, and the photocatalyst processing layer side substrate is removed from the microarray chip after energy irradiation, so that the microarray chip itself includes the photocatalyst processing layer. There is no. Therefore, it is possible to eliminate the semi-permanent action of the photocatalyst on the microarray chip.
[0207]
Hereinafter, the manufacturing method of the microarray chip of this embodiment in such a case will be described in detail for each step.
[0208]
(1) Microarray chip substrate preparation process
The substrate adjustment step for microarray chip in this embodiment is a decomposition removal layer that is decomposed and removed on the substrate by the action of a photocatalyst accompanying energy irradiation, and has a different contact angle with the liquid from the substrate, This is a step of adjusting the microarray chip substrate.
[0209]
The microarray chip substrate formed in this step will be described.
[0210]
(Base material for microarray chip)
a. Decomposition removal layer
First, the decomposition removal layer which comprises the base material for microarray chips is demonstrated. The decomposition / removal layer in this embodiment is a layer in which the decomposition / removal layer of the portion irradiated with energy is decomposed and removed by the action of the photocatalyst in the photocatalyst treatment layer during energy irradiation, and the decomposition / removal layer is decomposed and removed. It is a layer showing wettability different from that of the base material exposed at the time of exposure.
[0211]
Such a decomposed / removed layer has a pattern consisting of a portion having a decomposed / removed layer and a portion having no decomposed / removed layer without performing a development process or a cleaning process, that is, unevenness, because the portion irradiated with energy is decomposed and removed by the action of a photocatalyst. A pattern can be formed.
[0212]
This decomposition / removal layer is oxidatively decomposed and vaporized by the action of the photocatalyst by energy irradiation, and is therefore removed without any special post-treatment such as development / washing process. Depending on the material, a cleaning process or the like may be performed.
[0213]
In addition, this aspect is an aspect in which the fixing layer is patterned by using the decomposition / removal layer formed in a pattern and utilizing the wettability difference between the decomposition / removal layer and the base material. It is comprised so that the contact angle with respect to the liquid with a base material may differ.
[0214]
The wettability of such a decomposition / removal layer is that, when the immobilization layer is formed on the substrate where the decomposition / removal layer is decomposed and removed, it is decomposed and removed rather than the contact angle of the substrate with respect to the liquid. Increase the contact angle of the layer to the liquid.
[0215]
In such a case, the contact angle of the decomposition / removal layer with respect to the liquid having the surface tension equivalent to the surface tension of the applied fixing layer forming coating liquid is 1 ° or more, particularly 5 °, than that of the substrate. Above all, it is preferable that the angle is larger by 10 ° or more.
[0216]
In such a case, the water repellency required for the surface of the decomposition / removal layer is such that the contact angle with respect to a liquid having a surface tension equivalent to the surface tension of the applied fixing layer forming coating liquid is 30 ° or more, particularly It is preferably 40 ° or more, particularly 50 ° or more.
[0217]
Thus, if the decomposition removal layer has water repellency within the above range and the base material and the decomposition removal layer have a difference in wettability within the above range, the fixing layer forming coating solution is applied. This is because the immobilization layer can be accurately formed on the substrate without adhering to the decomposition removal layer.
[0218]
On the other hand, when the immobilization layer is formed on the decomposition removal layer, the contact angle of the decomposition removal layer with respect to the liquid is made smaller than the contact angle of the substrate with respect to the liquid. As a result, the immobilization layer forming coating liquid that forms the immobilization layer is less likely to adhere to the base material exposed by the decomposition removal layer being decomposed and removed, and to be satisfactorily adhered to the decomposition removal layer. Because you can.
[0219]
In addition, the contact angle with the liquid here is measured using a contact angle measuring instrument (Kyowa Interface Science Co., Ltd. CA-Z type) with a liquid having various surface tensions (from the microsyringe to the liquid. 30 seconds after dropping), and the result was obtained or the result was graphed. In this measurement, as a liquid having various surface tensions, a wetting index standard solution manufactured by Pure Chemical Co., Ltd. was used.
[0220]
In addition, the film thickness of the decomposition removal layer in the present embodiment is not particularly limited as long as it can be decomposed and removed by the action of the photocatalyst according to the energy to be irradiated in the pattern forming process described later. Specifically, the thickness is preferably 0.001 μm to 1 μm, particularly preferably in the range of 0.01 to 0.1 μm.
[0221]
The material that can be used for such a decomposition / removal layer is a material that is decomposed and removed by the above-described characteristics of the decomposition / removal layer, that is, the action of a photocatalyst accompanying energy irradiation, and preferably a liquid within the above range. A material having a contact angle of
[0222]
The method for forming the decomposition / removal layer is not particularly limited, but a general application such as spin coating is performed by selecting a material as described above and diluting it with an appropriate solvent to obtain a coating solution. The method of forming into a film using a method can be mentioned. Such a method for forming a thin film can be said to be an easy and cost-effective method. On the other hand, functional thin films, that is, film formation methods such as self-assembled monomolecular films, Langmuir-Brocket films, and alternating adsorption films can also be used. These methods are preferable because they enable the formation of a dense thin film with few defects.
[0223]
Here, the self-assembled monomolecular film, the Langmuir-Brocket film, and the alternating adsorption film used in this embodiment will be specifically described.
[0224]
(1) Self-assembled monolayer
Although the inventors do not know the existence of an official definition of self-assembled monolayer, the explanation of what is generally recognized as a self-assembled film is, for example, a review by Abraham Ulman. "Formation and Structure of Self-Assembled Monolayers", Chemical Review, 96, 1533-1554 (1996) is excellent. Referring to this review, a self-assembled monolayer can be said to be a monolayer formed as a result of adsorbing and binding (self-organizing) appropriate molecules to the appropriate substrate surface. Examples of materials capable of forming a self-assembled film include surfactant molecules such as fatty acids, organosilicon molecules such as alkyltrichlorosilanes and alkylalkoxides, organic sulfur molecules such as alkanethiols, and alkyl phosphates. And organic phosphoric acid molecules. The common commonality of the molecular structure is that there is a functional group that has a relatively long alkyl chain and interacts with the substrate surface at one molecular end. The portion of the alkyl chain is a source of intermolecular force when molecules are packed two-dimensionally. However, the example shown here has the simplest structure, having a functional group such as an amino group or a carboxyl group at the other end of the molecule, an alkylene chain part having an oxyethylene chain, or a fluorocarbon chain. Self-assembled monolayers composed of various molecules such as those of complex type chains have been reported. There is also a composite type self-assembled monolayer composed of a plurality of molecular species. In addition, recently, a polymer having a plurality of functional groups (which may have one functional group) or a linear polymer (which may have a branched structure) as represented by dendrimers has been developed. One layer formed on the surface of the substrate (the latter is collectively referred to as a polymer brush) may be considered as a self-assembled monolayer. In the present invention, these are also included in the self-assembled monolayer.
[0225]
(2) Langmuir-Blodgett membrane
If the Langmuir-Blodgett film used in this embodiment is formed on a substrate, there is no significant difference in form from the above-described self-assembled monolayer. It can be said that the Langmuir-Blodgett film is characterized by its formation method and the advanced two-dimensional molecular packing properties (high orientation and high order). That is, in general, Langmuir-Blodgett film-forming molecules are first developed on the gas-liquid interface, and the developed film is condensed by the trough to change into a highly packed condensed film. In practice, this is transferred to a suitable substrate and used. It is possible to form a monomolecular film to a multilayer film of an arbitrary molecular layer by the method outlined here. Further, not only low molecules but also polymers and colloidal particles can be used as the film material. Recent examples of the application of various materials are described in detail in the review by Tokuharu Miyashita et al. “Prospects for Nanotechnology for the Creation of Soft Nanodevices” Polymer Vol. 50, September, 644-647 (2001).
[0226]
(3) Alternate adsorption film
In general, an alternating adsorption film (Layer-by-Layer Self-Assembled Film) is made by adsorbing and bonding a material having at least two functional groups with positive or negative charges sequentially onto a substrate. It is a film formed by stacking. Since a material having a large number of functional groups has many advantages such as increased strength and durability of the membrane, recently, an ionic polymer (polymer electrolyte) is often used as a material. Further, particles having surface charges such as proteins, metals and oxides, so-called “colloid particles” are also frequently used as film-forming substances. More recently, membranes that actively utilize weaker interactions than ionic bonds such as hydrogen bonds, coordination bonds, and hydrophobic interactions have been reported. A relatively recent example of alternating adsorption films is slightly biased toward materials with electrostatic interaction as the driving force, but a review by Paula T. Hammond "Recent Explorations in Electrostatic Multilayer Thin Film Assembly" & Interface Science, 4, 430-442 (2000). The alternate adsorption film can be described by taking the simplest process as an example, by repeating the adsorption-washing of a material having a positive (negative) charge-adsorption-washing of a material having a negative (positive) charge a predetermined number of times. It is a film to be formed. As with Langmuir-Blodgett membranes, no development-condensation-transfer operations are required. Further, as apparent from the difference in these production methods, the alternating adsorption film generally does not have a two-dimensional high orientation / high order like the Langmuir-Blodgett film. However, the alternate adsorption film and its manufacturing method have advantages over conventional film formation methods, such as the ability to easily form a dense film without defects and the ability to form even fine irregular surfaces, tube inner surfaces, and spherical surfaces. Have many.
[0227]
b. Base material
Next, the base material of this embodiment will be described. In this aspect, since the immobilization layer is formed in a pattern using the above-described wettability difference between the decomposition removal layer and the substrate, when the immobilization layer is formed on the substrate, The base material which exhibits a contact angle smaller than the contact angle with respect to the liquid of the decomposition removal layer mentioned above is used. On the other hand, when the immobilization layer is formed on the above-described decomposition / removal layer, a substrate having a contact angle larger than the contact angle with respect to the liquid exhibited by the above-described decomposition / removal layer is used.
[0228]
(2) Pattern formation process
The pattern forming step in this embodiment is a photocatalyst processing layer side substrate in which a photocatalyst processing layer containing at least a photocatalyst is formed on the substrate, and the microarray chip substrate and the photocatalyst processing layer side substrate are converted into the photocatalyst. After disposing the treatment layer and the decomposition / removal layer so as to face each other and with a gap of 200 μm or less, the decomposition / removal layer is formed in a pattern by irradiating energy from a predetermined direction.
[0229]
Since this process is the same as that in the first embodiment described above, description thereof is omitted here.
[0230]
(3) Removal process
In this embodiment, after the energy irradiation, the photocatalyst processing layer substrate is removed from the microarray chip substrate. Thereby, the base material which has the decomposition removal layer formed in pattern shape on the base material is obtained.
[0231]
(4) Immobilization layer forming step
Next, a fixing layer forming step is performed. The immobilization layer forming step in this embodiment is a method of applying the immobilization layer forming coating liquid on the base material having the decomposition / removal layer formed in a pattern, and the wettability of the decomposition / removal layer and the base material. This is a step of forming an immobilization layer in a pattern on the decomposition / removal layer or on the base material where the decomposition / removal layer is decomposed and removed by utilizing the difference.
[0232]
In this embodiment, as described above, when the contact angle of the decomposition removal layer with respect to the liquid is smaller than that of the base material, the immobilization layer is formed on the decomposition removal layer. On the contrary, when the contact angle of the decomposition removal layer with respect to the liquid is larger than that of the base material, the fixation layer is formed on the exposed base material by the decomposition removal layer being decomposed and removed in the pattern forming process described above. The
[0233]
Other than that, the immobilization layer formed in this step is the same as that described in “D. Immobilization layer formation step” in the first embodiment described above, and thus the description thereof is omitted here.
[0234]
(5) Specific biopolymer immobilization process
The specific biopolymer immobilization step in this embodiment is a step of immobilizing the specific biopolymer on the immobilization layer formed by the above-described immobilization layer formation step.
[0235]
Since this step is the same as the “E. Specific biopolymer immobilization step” in the first embodiment described above, description thereof is omitted here.
[0236]
B. When using the decomposition removal layer as an immobilization layer
Next, the manufacturing method of the microarray chip of this aspect when the decomposition / removal layer that functions as the immobilization layer is used will be described.
[0237]
In such a case, the microarray chip manufacturing method according to the present embodiment includes forming a decomposition removal layer that is decomposed and removed by the action of a photocatalyst accompanying energy irradiation and functions as an immobilization layer on a base material. A microarray chip base material adjusting step for adjusting a material, and a photocatalyst processing layer side substrate in which a photocatalyst processing layer containing at least a photocatalyst is formed on the substrate, the microarray chip base material and the photocatalyst processing layer side substrate Is disposed so that the photocatalyst treatment layer and the decomposition removal layer face each other and with a gap of 200 μm or less, and then irradiated with energy from a predetermined direction, whereby the decomposition having a function as an immobilization layer From the pattern formation step of forming the removal layer in a pattern, and the microarray chip substrate It is characterized in that it has at least a removal step of removing the catalyst treatment layer side substrate.
[0238]
This mode in such a case will be described with reference to the drawings.
[0239]
FIG. 3 shows an example of a manufacturing method for manufacturing a microarray chip using a decomposition / removal layer having a function as an immobilization layer. First, a photocatalyst treatment layer side substrate 3 in which a photocatalyst treatment layer 2 made of at least a photocatalyst is formed on a substrate 1 is prepared. Further, separately from the photocatalyst processing layer side substrate 3, a base material 4 is prepared, and a decomposition removal layer 21 that is decomposed and removed by the action of the photocatalyst accompanying energy irradiation on the base material 4 and functions as an immobilization layer. Film formation is performed to prepare a microarray chip substrate 6 (see FIG. 3A).
[0240]
Furthermore, after the photocatalyst processing layer side substrate 3 and the microarray chip substrate 6 are arranged so that the photocatalyst processing layer 2 and the decomposition removal layer 21 face each other and have a predetermined gap, energy is passed through the photomask 7. Irradiate in a pattern (see FIG. 3B).
[0241]
Next, when the photocatalyst processing layer side substrate 3 is removed from the microarray chip substrate 6, the substrate 4 having the decomposition removal layer 21 formed in a pattern is obtained (see FIG. 3C). In this embodiment, since the decomposition / removal layer having the function as the immobilization layer is used, the decomposition / removal layer formed in a pattern like this can be used as the immobilization layer.
[0242]
Then, the microarray chip can be manufactured by attaching the specific biopolymer 11 on the decomposition / removal layer 21 having a function as an immobilization layer (see FIG. 3D).
[0243]
According to this aspect in this case, since the decomposition / removal layer having a function as an immobilization layer is used, the immobilization layer is also formed in a pattern by forming the decomposition / removal layer in a pattern. . Therefore, the labor required for patterning the immobilization layer is greatly simplified, so that the production efficiency can be improved. Also in this embodiment, the photocatalyst processing layer side substrate is removed from the microarray chip substrate after the decomposition / removal layer is irradiated with energy, and therefore the microarray chip does not include the photocatalyst processing layer. Therefore, the microarray chip of this embodiment can eliminate the influence of the action of the photocatalyst.
[0244]
The decomposition removal layer used for this aspect in such a case will be described. Regarding the microarray chip substrate preparation step, pattern formation step, removal step, and specific biopolymer formation step, “A. Difference in wettability between the degradation removal layer and the substrate in the second embodiment described above is used. Since it is the same as that described in “When a fixing layer is formed by use”, description thereof is omitted here.
[0245]
(Decomposition removal layer)
The material forming the decomposition / removal layer in this case is contained in the photocatalyst processing layer arranged in contact with the decomposition / removal layer or with a predetermined gap during the energy irradiation, that is, the energy irradiation. The material is not particularly limited as long as it is a material that can be decomposed and removed by the action of the photocatalyst and can fix the specific biopolymer. Specifically, the same material as the immobilization layer described in “D. Immobilization layer forming step” in the first embodiment described above can be used.
[0246]
As the film formation method of the decomposition removal layer, as described above, in addition to a general formation method such as forming a thin film by diluting such a material and applying it by a method such as spin coating, By forming using the above-described functional thin film, that is, a film forming method such as a self-assembled monomolecular film, a Langmuir-Brocket film, and an alternately adsorbing film, a dense thin film with few defects can be obtained. The self-assembled monomolecular film, the Langmuir-Brocket film, and the alternately adsorbed film are the same as described above, and a description thereof is omitted here.
[0247]
In this embodiment, such a decomposition / removal layer is formed in a pattern by being irradiated with energy after being arranged to face the photocatalyst treatment layer and having a predetermined gap. As a result, a patterned immobilization layer is obtained. A microarray chip can be manufactured by attaching and fixing a specific biopolymer on the decomposition / removal layer having a function as an immobilization layer.
[0248]
II. Microarray chip
Next, the microarray chip of the present invention will be described.
[0249]
The microarray chip of the present invention is characterized by using a decomposition removal layer that is decomposed and removed by the action of a photocatalyst accompanying energy irradiation. Furthermore, the microarray chip of the present invention includes a case where an immobilization layer is formed using such a difference in wettability between the decomposition removal layer and the substrate, and a case where the decomposition removal layer itself is used as the immobilization layer. Can be divided into Hereinafter, both cases will be referred to as a third embodiment and a fourth embodiment, and each embodiment will be described.
[0250]
1. Third embodiment
The microarray chip according to the third embodiment is manufactured using a decomposition / removal layer that exhibits wettability different from that of the substrate and is decomposed and removed by the action of a photocatalyst accompanying energy irradiation.
[0251]
Such a microarray chip of the third embodiment includes a base material, a decomposition removal layer formed in a pattern on the base material, and decomposed and removed by the action of a photocatalyst accompanying energy irradiation, and the decomposition removal layer or the The degradation removal layer is formed on the substrate which is decomposed and removed and is exposed, and an immobilization layer for immobilizing the biopolymer, and the target biopolymer immobilized on the immobilization layer and specific to the biopolymer Specific decomposition | disassembly removal layer and the said group differ in the contact angle with respect to the liquid.
[0252]
Such a microarray chip of this embodiment will be described with reference to the drawings.
[0253]
FIG. 4 shows an example of such a microarray chip in this embodiment. First, the decomposition removal layer 21 is formed on the base material 4 in a pattern. Here, in this aspect, since the wettability of the decomposition removal layer and the base material is different, the pattern due to the difference in wettability is formed by forming the decomposition removal layer in a pattern on the base material. Will be. The immobilization layer 10 is formed on the base material 4 where the decomposition / removal layer is decomposed and removed by utilizing the pattern due to the difference in wettability. FIG. 4 shows the case where the immobilization layer is formed on the base material, but the case where it is formed on the decomposition removal layer due to the wettability relationship between the base material and the decomposition removal layer. There may be. Furthermore, a specific biopolymer 11 is immobilized on the immobilization layer 10.
[0254]
In such a microarray chip in this embodiment, an immobilization layer is formed in a pattern, and a specific biopolymer is disposed thereon. Furthermore, since the immobilization layer is formed in a pattern using the difference in wettability between the decomposition removal layer and the base material, the region where the immobilization layer is not formed becomes a water-repellent region. ing. Therefore, for example, when a specific biopolymer is disposed on the immobilization layer by an ink jet method or the like, the specific biopolymer does not spread over the area of the immobilization layer, and the specific biopolymer It is possible to align the shapes of the two, and there is an advantage that the final detection sensitivity can be improved.
[0255]
In addition, since the base material, the decomposition removal layer, the immobilization layer, the specific biopolymer, and the like in this embodiment are the same as those described in the above-mentioned “I. Manufacturing method of microarray chip”, the description here is Omitted.
[0256]
2. Fourth embodiment
Next, the fourth embodiment will be described. The fourth embodiment is a microarray chip formed by using a decomposition removal layer that is decomposed and removed by the action of a photocatalyst accompanying energy irradiation and functions as an immobilization layer.
[0257]
Such a microarray chip in this embodiment includes a substrate, a decomposition removal layer that is formed in a pattern on the substrate, decomposes and removes by the action of a photocatalyst accompanying energy irradiation, and functions as an immobilization layer, and the decomposition It has a specific biopolymer which is immobilized on the removal layer and specifically binds to a target biopolymer.
[0258]
Further, the microarray chip of this embodiment will be described with reference to the drawings.
[0259]
FIG. 5 shows an example of the microarray chip of this embodiment. First, the base material 4 and the decomposition removal layer 21 are formed in a pattern on the base material 4. In this embodiment, since the decomposition / removal layer 5 has a function as an immobilization layer, a microarray chip is obtained by attaching and fixing the specific biopolymer 11 on the decomposition / removal layer 5. It can be.
[0260]
According to this aspect, by using the decomposition / removal layer that can be easily patterned only by energy irradiation, and using the decomposition / removal layer as the immobilization layer, the labor required for patterning the immobilization layer is greatly reduced. And the manufacturing process can be simplified. Moreover, since waste can be reduced in terms of material, it is advantageous in terms of cost.
[0261]
In addition, since the base material, the decomposition removal layer, the specific biopolymer, and the like in this embodiment are the same as those described in the above-mentioned “I. Microarray chip manufacturing method”, description thereof is omitted here.
[0262]
III. Playback method
In the present invention, after using the above-described microarray chip for detection or the like, only the portion to which the above-mentioned immobilization layer and specific biopolymer are attached is irradiated in a pattern via the above-mentioned photocatalyst processing layer side substrate. Can be reproduced.
[0263]
Since the microarray chip of the present invention is reproducible in this way, it can be used a plurality of times and has the advantage that the final cost can be reduced.
[0264]
FIG. 6 is a diagram for explaining an example of a method for reproducing a microarray chip according to the present invention. FIG. 6 (a) shows the microarray chip after use of detection of biopolymers and the like is finished. In the microarray chip regeneration method of the present invention, as shown in FIG. 6B, the immobilized layer 10 and the specific biopolymer 11 are arranged on such a used microarray chip. The energy such as ultraviolet light is arranged through the photomask 7 and the photocatalyst treatment layer side substrate 3 is disposed so that the photocatalyst treatment layer 2 and the specific biopolymer 11 face each other so that only a part is irradiated with energy. And irradiate energy. Thereby, the immobilization layer 10 and the specific biopolymer 11 are decomposed by the action of the photocatalyst contained in the photocatalyst processing layer 2. On the other hand, the portion not irradiated with energy remains as a water-repellent region as it is. In this case, the gap between the photocatalyst processing layer side substrate and the microarray chip, the energy irradiation method, and the like are the same as those described in “B. Pattern formation step” in the first embodiment described above. Further, if necessary, the substrate 4 in which the hydrophilic region 8 is formed in a pattern on the wettability changing layer 5 is obtained as shown in FIG.
[0265]
By performing the above-described steps shown in FIGS. 1D and 1E on such a substrate, it can be reused as a microchip array.
[0266]
FIG. 7 shows another example of the reproducing method of the microchip array of the present invention. In this microchip array, a microarray chip side light-shielding portion 33 is formed between the base material 4 and the wettability changing layer 5, and the microarray chip side light-shielding portion 33 delimits a portion where the immobilization layer is formed. (See FIG. 7A). When reproducing such a microarray chip, as shown in FIG. 7B, the photocatalyst processing layer side substrate 3 is arranged so that the photocatalyst processing layer 2 and the specific biopolymer 11 face each other. 4 is irradiated with energy from the surface side where the wettability changing layer 5 is not formed, so that the portion where the microarray chip side light-shielding portion 33 is not formed, that is, the immobilization layer 10 and the specific biopolymer 11 are attached. Energy is irradiated only to the part that is being removed, and these are removed by the action of the photocatalyst contained in the photocatalyst treatment layer 2, and the wettability changing layer 5 is patterned in a pattern as shown in FIG. The base material 4 in which the hydrophilic region 8 is formed is obtained. At this time, the arrangement of the photocatalyst treatment layer side substrate and the energy irradiation are the same as those described in “B. Pattern formation method” in the first embodiment described above. Is omitted.
[0267]
In addition, the microarray chip-side light-shielding portion in this aspect is a member that plays a role of hindering energy transmission during energy irradiation, and the same material as the above-described photocatalyst processing layer-side light-shielding layer can be used. Therefore, since the microarray chip side light shielding portion is the same as the photocatalyst processing layer side light shielding portion described above, description thereof is omitted here.
[0268]
Also in this case, similar to the above-described example, this substrate can be reused as a microchip array by performing the above-described steps shown in FIGS. 1D and 1E.
[0269]
As such a regeneration method, FIG. 6 and FIG. 7 show the case of the microarray chip formed using the wettability changing layer, but the case of the microarray chip formed using the decomposition removal layer is also shown. It can be played back by the same playback method.
[0270]
The substrate, wettability changing layer, photocatalyst treatment layer side substrate, immobilization layer, specific biopolymer, etc. in the microarray chip regeneration method described above are those described in “I. Microarray chip manufacturing method” described above. The description here is omitted.
[0271]
The present invention is not limited to the above embodiment. The above-described embodiment is an exemplification, and the present invention has any configuration that has substantially the same configuration as the technical idea described in the claims of the present invention and that exhibits the same effects. Are included in the technical scope.
[0272]
【Example】
The following examples further illustrate the invention.
[Example 1]
1. Formation of photocatalyst treatment layer side substrate
5 g of trimethoxymethylsilane (GE Toshiba Silicone Co., Ltd., TSL8113) and 2.5 g of 0.5N hydrochloric acid were mixed and stirred for 8 hours. This was diluted 10 times with isopropyl alcohol to obtain a primer layer composition.
[0273]
The primer layer composition was applied onto a photomask substrate by a spin coater and dried at 150 ° C. for 10 minutes to form a transparent primer layer (thickness 0.2 μm).
[0274]
Next, 30 g of isopropyl alcohol, 3 g of trimethoxymethylsilane (manufactured by GE Toshiba Silicone Co., Ltd., TSL8113) and 20 g of ST-K03 (manufactured by Ishihara Sangyo Co., Ltd.), which is a photocatalytic inorganic coating agent, are mixed at 100 ° C. Stir for 20 minutes. This was diluted 3 times with isopropyl alcohol to obtain a composition for a photocatalyst treatment layer.
[0275]
A transparent photocatalyst treatment layer (thickness: 0.15 μm) is formed by applying the composition for a photocatalyst treatment layer on a photomask substrate on which a primer layer is formed by a spin coater and performing a drying treatment at 150 ° C. for 10 minutes. Formed.
[0276]
2. Preparation of substrate for microarray chip
30 hours of isopropyl alcohol, 0.4 g of fluoroalkylsilane (manufactured by GE Toshiba Silicone), 3 g of trimethoxymethylsilane (manufactured by GE Toshiba Silicone, TSL8113) and 2.5 g of 0.5 N hydrochloric acid are mixed for 8 hours. Stir. This was diluted 100 times with isopropyl alcohol to obtain a wettability changing layer composition.
[0277]
The said composition for wettability change layers was apply | coated with the spin coater on the glass substrate, and the transparent wettability change layer (thickness 0.1 micrometer) was formed by performing the drying process for 10 minutes at 150 degreeC.
[0278]
3. Confirmation of hydrophilic region formation by exposure
The photocatalyst treatment layer side substrate and the wettability changing layer are opposed to each other with a gap of 100 μm, and 40 mW / cm from the photomask side by an ultrahigh pressure mercury lamp (wavelength 365 nm).²The region where the immobilization layer on the wettability changing layer is formed was defined as a hydrophilic region.
[0279]
At this time, a contact angle measuring device (Kyowa Interface Science Co., Ltd.) is used to determine the contact angle between a liquid repellent region and a hydrophilic region consisting of unexposed areas and a wetting index standard solution (manufactured by Junsei Chemical Co., Ltd.) having a surface tension of 40 mN / m As a result of measurement using a CA-Z type) (30 seconds after dropping a droplet from a microsyringe), they were 70 degrees and 7 degrees, respectively.
[0280]
4). Formation of immobilization layer
The substrate was immersed in a 3% aqueous solution of poly L-lysine and pulled up to form an immobilization layer only in the hydrophilic region.
[0281]
5. Fabrication of microarray chip
A microarray chip was produced by discharging various DNA solutions onto the immobilization layer using an ink jet apparatus and immobilizing them.
[0282]
6). Playback method
The photocatalyst processing layer side substrate 1 and the microarray chip are aligned to face each other with a gap of 100 μm, and 40 mW / cm from the photocatalyst processing layer side substrate by an ultrahigh pressure mercury lamp (wavelength 365 nm).²It was exposed for 300 seconds at an illuminance of, and regenerated.
[Example 2]
1. Formation of photocatalyst treatment layer side substrate
5 g of trimethoxymethylsilane (GE Toshiba Silicone Co., Ltd., TSL8113) and 2.5 g of 0.5N hydrochloric acid were mixed and stirred for 8 hours. This was diluted 10 times with isopropyl alcohol to obtain a primer layer composition.
[0283]
The primer layer composition was applied onto a photomask substrate by a spin coater and dried at 150 ° C. for 10 minutes to form a transparent primer layer (thickness 0.2 μm).
[0284]
Next, 30 g of isopropyl alcohol, 3 g of trimethoxymethylsilane (manufactured by GE Toshiba Silicone Co., Ltd., TSL8113) and 20 g of ST-K03 (manufactured by Ishihara Sangyo Co., Ltd.), which is a photocatalytic inorganic coating agent, are mixed at 100 ° C. Stir for 20 minutes. This was diluted 3 times with isopropyl alcohol to obtain a composition for a photocatalyst treatment layer.
[0285]
A transparent photocatalyst treatment layer (thickness: 0.15 μm) is formed by applying the composition for a photocatalyst treatment layer on a photomask substrate on which a primer layer is formed by a spin coater and performing a drying treatment at 150 ° C. for 10 minutes. Formed.
[0286]
2. Formation of immobilization layer
The glass substrate was dipped in a 3% aqueous solution of poly L-lysine and pulled up to form an immobilization layer.
[0287]
3. Patterning of immobilization layer by exposure
The photocatalyst processing layer side substrate and the fixing layer are opposed to each other with a gap of 100 μm, and 40 mW / cm from the photomask side by an ultrahigh pressure mercury lamp (wavelength 365 nm).²The immobilization layer was patterned by exposing for 180 seconds at an illuminance of.
[0288]
4). Fabrication of microarray chip
A microarray chip was produced by discharging various DNA solutions onto the immobilization layer using an ink jet apparatus and immobilizing them.
[0289]
6). Playback method
The photocatalyst treatment layer side substrate and the microarray chip are opposed to each other with a gap of 100 μm, and 40 mW / cm from the photocatalyst treatment layer side substrate by an ultrahigh pressure mercury lamp (wavelength 365 nm).²It was exposed for 300 seconds at an illuminance of, and regenerated.
[0290]
【The invention's effect】
According to the present invention, after the energy irradiation, the photocatalyst processing layer side substrate is removed from the microarray chip substrate, so that the microarray chip itself does not include the photocatalyst processing layer. There is no worry about the effect. Furthermore, since the distance between the photocatalyst treatment layer and the characteristic change layer is within the above-described range, a pattern can be formed efficiently and with a good difference in characteristics. Furthermore, since the immobilization layer is formed using the change pattern of the characteristic change layer formed by energy pattern irradiation, a high-definition pattern can be obtained. Therefore, a high-definition microarray chip can be obtained by immobilizing a specific biopolymer on the immobilization layer.
[Brief description of the drawings]
FIG. 1 is a process chart showing an example of a method for producing a microarray chip of the present invention.
FIG. 2 is a process diagram showing another example of a method for manufacturing a microarray chip of the present invention.
FIG. 3 is a process diagram showing another example of a method for producing a microarray chip of the present invention.
FIG. 4 is a schematic cross-sectional view showing an example of a microarray chip of the present invention.
FIG. 5 is a schematic cross-sectional view showing another example of the microarray chip of the present invention.
FIG. 6 is a process diagram for explaining an example of a microarray chip reproduction method of the present invention.
FIG. 7 is a process diagram for explaining another example of the microarray chip reproduction method of the present invention.
FIG. 8 is a schematic cross-sectional view showing an example of the photocatalyst processing layer side substrate of the present invention.
FIG. 9 is a schematic sectional view showing another example of the photocatalyst processing layer side substrate of the present invention.
FIG. 10 is a schematic sectional view showing another example of the photocatalyst processing layer side substrate of the present invention.
FIG. 11 is a schematic cross-sectional view showing another example of the photocatalyst processing layer side substrate of the present invention.
[Explanation of symbols]
1 ... Substrate
2… Photocatalyst treatment layer
3 ... Photocatalyst processing layer side substrate
4 ... Base material
5… wettability change layer
6 ... Base material for microarray chip
7 ... Photomask
8… hydrophilic region
9 ... Water repellent area
10: Immobilization layer
11… Specific biopolymer

Claims (32)

A microarray chip base material adjusting step for adjusting a microarray chip base material having a characteristic change layer whose characteristics are changed by the action of a photocatalyst associated with energy irradiation;
Using a photocatalyst treatment layer side substrate on which a photocatalyst treatment layer containing at least a photocatalyst is formed on the substrate, the photocatalyst treatment layer and the characteristic change layer face each other with the microarray chip substrate and the photocatalyst treatment layer side substrate. And a pattern forming step of forming a pattern due to a difference in characteristics on the surface of the characteristic change layer by irradiating energy from a predetermined direction after being arranged with a gap of 200 μm or less;
A removal step of removing the photocatalyst processing layer side substrate from the microarray chip substrate,
An immobilization layer forming step of forming an immobilization layer in a pattern according to a pattern due to a difference in characteristics of the characteristic change layer,
A specific biopolymer immobilization step of immobilizing a specific biopolymer on the immobilization layer;
A method for producing a microarray chip, comprising: A microarray chip substrate adjustment step of adjusting a microarray chip substrate having a wettability changing layer in which wettability is changed by the action of a photocatalyst associated with energy irradiation;
Using a photocatalyst treatment layer side substrate in which a photocatalyst treatment layer containing at least a photocatalyst is formed on the substrate, the microarray chip substrate and the photocatalyst treatment layer side substrate are formed by the photocatalyst treatment layer and the wettability changing layer. A pattern forming step of forming a pattern due to a difference in wettability on the surface of the wettability changing layer by irradiating energy from a predetermined direction after being arranged with a gap of 200 μm or less facing each other;
A removal step of removing the photocatalyst processing layer side substrate from the microarray chip substrate,
An immobilization layer forming step of forming an immobilization layer on the wettability change layer according to the pattern of the wettability change layer;
A specific biopolymer immobilization step of immobilizing a specific biopolymer on the immobilization layer;
A method for producing a microarray chip, comprising: The method for producing a microarray chip according to claim 2, wherein the wettability changing layer is a layer containing an organopolysiloxane. The method for producing a microarray chip according to claim 3, wherein the organopolysiloxane is a polysiloxane containing a fluoroalkyl group. The organopolysiloxane is Y _n SiX _(4-n) (where Y represents an alkyl group, a fluoroalkyl group, a vinyl group, an amino group, a phenyl group or an epoxy group, and X represents an alkoxyl group or a halogen. 4. n is an integer from 0 to 3.) It is an organopolysiloxane which is one or two or more hydrolytic condensates or co-hydrolytic condensates of a silicon compound represented by Or the manufacturing method of the microarray chip | tip of Claim 4. The said microarray chip base material consists of a base material and the wettability change layer currently formed on the said base material, The claim in any one of Claim 2-5 characterized by the above-mentioned. Manufacturing method of microarray chip. The method for producing a microarray chip according to claim 2, wherein the substrate for microarray chip comprises a wettability changing layer having self-supporting property. A base material for microarray chip is prepared by forming a base material for microarray chip that is decomposed and removed by the action of a photocatalyst accompanying energy irradiation and having a contact angle with a liquid different from that of the base material. Adjustment process;
Using a photocatalyst treatment layer side substrate on which a photocatalyst treatment layer containing at least a photocatalyst is formed, the photocatalyst treatment layer and the decomposition / removal layer face the microarray chip substrate and the photocatalyst treatment layer side substrate. And a pattern forming step of forming the decomposition removal layer in a pattern by irradiating energy from a predetermined direction after being arranged with a gap of 200 μm or less,
A removal step of removing the photocatalyst processing layer side substrate from the microarray chip substrate,
According to the pattern of the decomposition removal layer, an immobilization layer forming step of forming an immobilization layer on the decomposition removal layer or the substrate;
A specific biopolymer immobilization step of immobilizing a specific biopolymer on the immobilization layer;
A method for producing a microarray chip, comprising: On the substrate, a microarray chip base material adjusting step for adjusting the microarray chip base material by forming a decomposition removal layer that is decomposed and removed by the action of the photocatalyst accompanying energy irradiation and functions as an immobilization layer;
Using a photocatalyst treatment layer side substrate on which a photocatalyst treatment layer containing at least a photocatalyst is formed, the photocatalyst treatment layer and the decomposition / removal layer face the microarray chip substrate and the photocatalyst treatment layer side substrate. And a pattern forming step of forming the decomposition / removal layer having a function as an immobilization layer in a pattern by irradiating energy from a predetermined direction after being arranged with a gap of 200 μm or less,
A removal step of removing the photocatalyst processing layer side substrate from the microarray chip substrate,
A specific biopolymer immobilization step of immobilizing a specific biopolymer on the decomposition removal layer;
A method for producing a microarray chip, comprising: 10. The method of manufacturing a microarray chip according to claim 8, wherein the decomposition / removal layer is a self-assembled monomolecular film, a Langmuir-Blodgett film, or an alternating adsorption film. The microarray according to any one of claims 1 to 10, wherein the photocatalyst processing layer side substrate includes a substrate and a photocatalyst processing layer formed in a pattern on the substrate. Chip manufacturing method. The photocatalyst treatment layer side substrate comprises a substrate, a photocatalyst treatment layer formed on the substrate, and a photocatalyst treatment layer side light-shielding portion formed in a pattern, and energy irradiation in the pattern formation step is a photocatalyst treatment. The method for manufacturing a microarray chip according to any one of claims 1 to 10, wherein the method is performed from a layer side substrate. The photocatalyst treatment layer side substrate is formed with a photocatalyst treatment layer on the substrate, and the photocatalyst treatment layer side light-shielding portion is formed in a pattern on the photocatalyst treatment layer. Manufacturing method of microarray chip. 13. The photocatalyst treatment layer side substrate, wherein the photocatalyst treatment layer side light shielding portion is formed in a pattern on the substrate, and the photocatalyst treatment layer is further formed thereon. Manufacturing method of microarray chip. 15. The photocatalyst processing layer side substrate is formed by forming a photocatalyst processing layer through a primer layer on a photocatalyst processing layer side light-shielding portion formed in a pattern on a transparent substrate. The manufacturing method of the microarray chip | tip of description. In the photocatalyst treatment layer side substrate, a spacer having a thickness in the range of 0.2 μm to 10 μm is formed in a pattern on the photocatalyst treatment layer, and the spacer and the property change layer, the wettability change layer or The method for manufacturing a microarray chip according to any one of claims 1 to 10, wherein the energy is irradiated while contacting the decomposition removal layer. 17. The method of manufacturing a microarray chip according to claim 16, wherein the spacer is a photocatalyst processing layer side light shielding portion made of a light shielding material. The method of manufacturing a microarray chip according to any one of claims 1 to 17, wherein the photocatalyst processing layer is a layer made of a photocatalyst. 19. The method of manufacturing a microarray chip according to claim 18, wherein the photocatalyst processing layer is a layer formed by depositing a photocatalyst on a substrate by a vacuum film forming method. The method of manufacturing a microarray chip according to any one of claims 1 to 17, wherein the photocatalyst treatment layer is a layer having a photocatalyst and a binder. The photocatalyst is composed of titanium oxide (TiO ₂ ), zinc oxide (ZnO), tin oxide (SnO ₂ ), strontium titanate (SrTiO ₃ ), tungsten oxide (WO ₃ ), bismuth oxide (Bi ₂ O _3). The microarray chip according to any one of claims 1 to 20, wherein the microarray chip is one or more substances selected from iron oxide (Fe ₂ O ₃ ). Production method. The method of manufacturing a microarray chip according to claim 21, wherein the photocatalyst is titanium oxide (TiO ₂ ). When irradiating energy in the pattern formation step, the distance between the photocatalyst treatment layer and the property change layer, the wettability change layer, or the decomposition removal layer is set within a range of 0.2 μm to 10 μm. The method of manufacturing a microarray chip according to any one of claims 1 to 22, wherein: The method of manufacturing a microarray chip according to any one of claims 1 to 23, wherein the energy irradiation is performed while heating the photocatalyst processing layer. The method for manufacturing a microarray chip according to any one of claims 1 to 24, wherein the immobilization layer is formed of a substance having a cationic property. The method for producing a microarray chip according to any one of claims 1 to 25, wherein the specific biopolymer is a probe DNA that specifically binds to a target nucleic acid. . 27. The microarray chip side light-shielding portion is formed on the substrate so as to divide a portion where the immobilization layer is formed. 27. The claim according to any one of claims 1 to 26, wherein: Manufacturing method of microarray chip. A substrate, a decomposition removal layer formed in a pattern on the substrate and decomposed and removed by the action of a photocatalyst upon energy irradiation, and a group on which the decomposition removal layer or the decomposition removal layer is decomposed and removed and exposed An immobilization layer for immobilizing a biopolymer formed on a material, and a specific biopolymer that is immobilized on the immobilization layer and specifically binds to a target biopolymer. The microarray chip is characterized in that the decomposition removal layer and the substrate have different contact angles with respect to the liquid. A substrate, a decomposition formed on the substrate in a pattern, decomposed and removed by the action of a photocatalyst upon energy irradiation, and functioned as an immobilization layer; and the target immobilized on the decomposition / removal layer A microarray chip comprising a specific biopolymer that specifically binds to the biopolymer to be obtained. 30. The microarray chip according to claim 28 or 29, wherein the decomposition / removal layer is any of a self-assembled monolayer, a Langmuir-Blodgett film, and an alternating adsorption film. The microarray chip according to any one of claims 28 to 30, wherein the immobilization layer is formed of a substance having a cationic property. After using the microarray chip manufactured by the method for manufacturing a microarray chip according to any one of claims 1 to 27, a photocatalyst processing layer containing at least a photocatalyst is formed on the substrate. A method for regenerating a microarray chip, wherein energy is irradiated in a pattern only to a portion where the specific biopolymer is attached through the photocatalyst treatment layer side substrate.

Images