JP2015512984A - 基材上にタンパク質の微細構造グラフティングを行うための装置 - Google Patents
基材上にタンパク質の微細構造グラフティングを行うための装置 Download PDFInfo
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- JP2015512984A JP2015512984A JP2014561457A JP2014561457A JP2015512984A JP 2015512984 A JP2015512984 A JP 2015512984A JP 2014561457 A JP2014561457 A JP 2014561457A JP 2014561457 A JP2014561457 A JP 2014561457A JP 2015512984 A JP2015512984 A JP 2015512984A
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Abstract
Description
−前記マトリクスは、前記光を反射によって伝播するマイクロミラーのマトリクスであり、
−前記光学システムは顕微鏡対物レンズであり、
−前記光源は紫外線波長で発光するレーザであり、
−前記紫外線波長は365nmである。
−第一の容器に、ベンゾフェノンと第一のタンパク質を含む第一の水溶液を満たすステップと、
−前記微少流体回路に前記第一の水溶液を満たして、第一の溶液と前記層が前記開口部において接触するようにするステップと、
−前記光によって、前記第一の構造化パターンの前記第一の微細構造画像を層の上に形成して、前記第一のタンパク質を層の上にフォトプリントするステップと、
を含む。
−前記第一のタンパク質は蛍光性である。
−第二の容器に、ベンゾフェノンと第二のタンパク質を含む第二の水溶液を満たすステップと、
−前記第一の水溶液を前記第二の水溶液と交換して、第二の溶液と前記層が前記開口部において接触するようにするステップと、
−前記第一の構造化パターンを前記第二の構造化パターンと交換して、前記第二のタンパク質を層の上にフォトプリントするステップと、
を含む上記の方法にも関する。
−前記第二のタンパク質は蛍光性である。
−微少流体回路に、第一のタンパク質を含む第一の水溶液を、第一の溶液とPEGの層が基材上に堆積されたPEG層によって覆われるマイクロチャネル開口部において接触するまで満たすステップと、
−前記光によって層の上に、第一の構造化パターンの、顕微鏡対物レンズによって縮小された第一の微少構造画像を形成して、第一のタンパク質を層の上にフォトプリントするステップと、
を含む方法によって得られる。
−光学的コントラストを持つパターンで光を伝播する、マイクロミラー以外のあらゆるマトリクスが本発明に適している。光の反射または吸収ではなく透過によって動作する液晶空間変調器が本発明に適しているであろう。
−パターンを作り、層の上にその画像を生成するあらゆる手段が本発明に適している。コントラストのあるパターンまたは発光パターンが光透過により生成される光源光透過型システムもまた本発明に適しており、このパターンは対物レンズによって再現され、対物レンズによって基材の第一の表面上に結像される。したがって、本発明の第一の実施形態の、マイクロミラーのマトリクスの反射による動作と倒立顕微鏡対物レンズとの組み合わせは、本発明の目的のための照明手段または光源光の空間変調器の一例である。
−システムが光学的および寸法的に安定しており、取り外しが不要であり、その一方で、本願における本質として、印刷層としてのPEGと溶液内のベンゾフェノンの使用を通じて、タンパク質の非特異的グラフティングを限定することが可能である。
−単純にマイクロバルブの数を増やし、ある時点で1つのマイクロバルブだけが開き、他はすべて閉じているようにする、これらのマイクロバルブの開放ロジックを使用することによって、所望の容器数と複数のタンパク質に適応させることができる。したがって、溶液中に各タンパク質とともに各タンパク質をグラフティングする手段を含めることが可能であり、そのグラフィング手段が基材に適しているという条件が満たされれば、本発明により基材上に所望の数のタンパク質を印刷することが可能であり、単一タンパク質の印刷と比較してもコントラストが低下せず、時間の損失もない。ロジックは、具体的にはコンピュータによって制御でき、本発明による印刷システムをできるだけ自動化できる。
−随意選択により、すすぎラインを設けずに動作できる。実際に、本発明の第一の実施形態の説明から、層上の溶液の流れが、照明がない場合に活性であれば、第一の表面の洗浄に寄与することが明らかである。この洗浄は、第一の実施形態の第一から第二の構成へと切り替わる時の、第二の流体による第一の流体の洗浄にも当てはまる。切替中、すなわち第一の表面と接触する流体の交換時、すなわち切り替え段階で、流体混合物が第一の表面と接触するため、タンパク質の混合物の付着または印刷を回避するために照明をオフにすることが望ましい可能性がある。照明は、印刷に望まれる精度の純粋な流体だけが第一の表面と接触していると考えられた時、すなわち、第一の表面上の単一タンパク質にとって望まれるケミカルコントラストが実現された時に、再びオンにすることができる。当業者であれば、印刷される各タンパク質について得られるコントラストに従って間欠的照明を容易に決定できる。
−前記第一の水溶液を前記緩衝液と、洗浄手段を通じて交換するサブステップと、
−緩衝液を第二の水溶液と交換して、第二の溶液と前記層を前記開口部において接触させるサブステップと、
を含んでいてもよいことがわかる。
−各タンパク質i(iは1〜N)に関して、
・少なくとも1つの光活性化可能な分子を含む水溶液という意味で光活性化可能な水溶液を含み、水溶液がまたタンパク質iを含むような容器を制御するマイクロバルブを開き、前記層がタンパク質iの水溶液と接触できるようにするサブステップと、
・光源、具体的には紫外線(UV)光源によって層を照明することによってパターンiを印刷し、タンパク質iを層の上に印刷できるようにするサブステップと、
・光源からの照明をやめるサブステップと、
・マイクロバルブiを閉じるサブステップと、
・洗浄手段を介してタンパク質iの水溶液の層を洗浄するための緩衝液を収容する緩衝液容器を制御する緩衝液マイクロバルブを開くサブステップと、
を実行するステップを含んでいてもよいことがわかる。
Claims (9)
- 基材上にいくつかのタンパク質の微細構造グラフティングを行う装置において、
基材(7)と、層と、マトリクスと、光源(9)と、光学システムと、第一の水溶液を受ける第一の容器(1)と、第二の水溶液を受ける第二の容器(2)と、微少流体回路と、を含み、
膜が基材上に配置され、
光源がマトリクスを光で照明するのに適しており、
マトリクスが光を第一の構造化パターンで伝播するのに適しており、
マトリクスが第一の構造化パターンを第二の構造化パターンと交換するための光学的手段を含み、
光学システムが層上に、第一のパターンの第一の微細構造画像を形成するのに適しており、
回路が第一の水溶液を収容するのに適しており、
回路が、その開口部において第一の溶液を層と接触させるための開口部を含み、
回路が第一の溶液を第二の溶液と交換するための微少流体手段を含み、
層がポリエチレングリコールを含む
ことを特徴とする装置。 - 請求項1に記載の装置において、
前記マトリクスは、光を反射により伝播するマイクロミラーのマトリクス(10)であることを特徴とする装置。 - 請求項1または2に記載の装置において、
前記光学システムが顕微鏡対物レンズ(11)であることを特徴とする装置。 - 請求項1〜3のいずれか1項に記載の装置において、
前記光源が紫外線波長で発光するレーザであることを特徴とする装置。 - 請求項4に記載の装置において、
前記紫外線波長が365nmであることを特徴とする装置。 - 請求項1〜5のいずれか1項に記載の装置を使用して基材上にタンパク質の微細構造グラフティングを行う方法において、
−第一の容器に、ベンゾフェノンと第一のタンパク質を含む第一の水溶液を満たすステップと、
−前記微少流体回路に前記第一の水溶液を満たして、第一の溶液と前記層を前記開口部において接触させるステップと、
−前記光により、前記第一の構造化パターンの前記第一の微細構造画像を層の上に形成し、前記第一のタンパク質を層の上にフォトプリントするステップと、
を含むことを特徴とする方法。 - 請求項6に記載の方法において、
前記第一のタンパク質が蛍光性であることを特徴する方法。 - 請求項6または7に記載の方法において、
−第二の容器に、ベンゾフェノンと第二のタンパク質を含む第二の水溶液を満たすステップと、
−前記第一の水溶液を前記第二の水溶液と交換して、第二の溶液と前記層を前記開口部において接触させるステップと、
−前記第一の構造化パターンの前記第二の構造化パターンと交換して、前記第二のタンパク質を層の上にフォトプリントするステップと、
を含むことを特徴とする方法。 - 請求項6〜8のいずれか1項に記載の方法において、
前記第二のタンパク質が蛍光性であることを特徴とする方法。
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