JP4758891B2 - 微小流体デバイス上の加熱、冷却および熱サイクリングのためのシステムおよび方法 - Google Patents
微小流体デバイス上の加熱、冷却および熱サイクリングのためのシステムおよび方法 Download PDFInfo
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Description
(発明の分野)
本発明は、熱サイクリングにおける使用のための微小流体デバイス上の一体型ヒーターおよびクーラーに、より詳細には、加熱、冷却および熱サイクリングシステムを搭載した携帯用微小流体カードに関する。本発明は、さらに、例えば、PCRまたはrtPCRでの使用のための熱サイクリングを提供するために、微小流体デバイスと組み合わせた使用のための一体型熱交換回路、または熱電気冷却器(TEC)を有する微小流体カードに関する。
液体のナノリットルサイズの容量に対する制御を提供する一体型微小流体取り扱いシステムは、現在の分析試験を小型化すること、および生物医学的試験で頻繁に用いられる小サンプルサイズを取り扱うことの両方で有用である。全体の化学的分析は、単一の微小流体デバイス上で実施され得る。これら微小流体デバイスは、チャネル、バルブ、ポンプ、フローセンサー、混合チャンバー、光学的検出器のような構成要素を含む。これら構成要素およびシステムの例は、米国特許第5,932,100号;同第5,922,210号;同第6,387,290号;同第5,747,349号;同第5,748,827号;同第5,726,751号;同第5,724,404号;同第5,716,852号;同第5,974,867号;同第6,007,775号;同第5,972,710号;同第5,971,158号;同第5,948,684号;および同第6,171,865号(これらの特許は、それらの全体が本明細書によって参考として援用される)中に見出され得る。
本発明は、一般に、加熱、冷却および熱サイクリングシステムを搭載した微小流体デバイス、ならびに一体型熱交換器回路または熱電気冷却器(TEC)を有する微小流体デバイスに関する。
上記のように、本発明は、一般に、オンボード加熱、冷却および熱サイクリングシステムを備えた微小流体デバイス、熱サイクリングを提供するために微小流体デバイスと組み合わせて用いられる一体型熱交換器回路またはTECを有する微小流体デバイスに関する。
本発明の別の局面に従って、上記一体化熱交換器は、独立して加熱されるかもしくは冷却されるかのいずれかの流体を含有する微小流体回路であるか、またはアッセイ流体を含有する微小流体回路に隣接して配置される発熱性材料もしくは吸熱性材料であり、それによって、回路に隣接する流体は、独立の回路中にあるアッセイ流体に温度変化を与える。上記熱交換器回路および上記アッセイ含有回路の両方が、上記微小流体カード上に備えられる。上記熱交換器回路中の流体は、上記カードを機器の多岐管に連結することによって循環されて、ポンプ輸送手段を提供し得る。
ColdSrc−冷流体貯蔵タンク(この場合、約0.3℃の氷水)における冷流体の温度を示す。
本発明のさらに別の代替的実施形態において、熱サイクリングは、熱電気冷却器(TEC)(例えば、ペルチェ)を用いて達成され得る。図10は、以下にさらに記載される本発明の熱電気サイクラーの構成要素のフロー概略図を示す。この配置は、PCRおよびrtPCRに伴う使用のための微小流体増幅チャンバーのための加熱冷却源として、TECを使用することの可能性を試験するために使用された。
電源 0〜20vdc(7.5VDCに設定)
逆電流方向へのDPDTスイッチ
熱シンク
DVM
TEC(Melcor CPO−8−63−06MM、12mm×25mm、Imax 2.1A、Vmax 7.62vdc)
熱電対
Micronics「run motor」ソフトウェアおよびThermocycler Dart(データ取得用)。
PCを介した通信
・RS−232
・USB
・GPIB
サーミスターセンサー 20℃〜100℃
TECを温度まで上昇させる能力およびTECを温度まで低下させる能力。
調節可能電圧出力 0〜20vdc、電流制限あり(または別個の電源を使用する能力)
データを取得および収集する能力。
P=1℃〜200℃
I=1秒以下
D=1秒以下
加熱冷却のために異なるPID回路を使用する能力。
TECを、熱シンクとして作用するステンレス鋼テーブル上に配置した。熱電対を、上記TECの上面にテープで付けた。データを、TECが高温から低温にサイクルした際にとった。この試験により、60℃から95℃になるのに4.25秒という遷移時間、または8.65℃/秒を示したデータを得た。冷却時間は、96℃から60℃になるのに3秒、または12℃/秒であった。
簡素な積層カードを、増幅チャンバーが0.004”Mylarの1つの層でキャッピングされるように設計した。これは、チャンバーのキャッピング層がTECと直接接触して配置されることを可能にした。図11に示すように、熱電対を、上記増幅チャンバーに挿入し、そしてそのチャンバーを、Fluorinert FC−40で充填した。
TECは、熱を片側からもう一方の側へ移動させる;プロセスにおいて、TECは熱を加える(TECは、かなりわずかな電流を起こす)。冷たい側が、すでに冷却した表面に接する場合、その表面から移動される熱は、最小であり、「熱い」側で起こった加熱は、主に、TECを通った電流に由来する。これは、TECが、冷たいステンレス鋼テーブル(約17℃)と直接接触した第一の試験において明らかである。何回かのサイクルの後、TECの下の領域は、わずかに熱くなり、そして70℃から95℃への上昇時間は、より速い。
16秒というサイクル時間(試験において最も悪い例)は、適切なサイズの熱シンク、TEC、およびより効率的な熱インターフェース材料を使用して大きく改善され得る。たとえ16秒においても、30回の完全なサイクルは、8分かかるだけである。TECは、上記カードの増幅領域に適合する大きさにされる。
上記のサイクル試験を、Parker Chomerics Thermagap材料61−02−0404−F574(0.020”の厚さ)を使用して繰り返した。574シリーズは、1.6W/m−Kの熱伝導率を提供するのにほんの5〜10psiの圧力を必要とする軟性エラストマー(<5Shore A)である。
図18に示されるように、熱サイクルグラフィックインターフェースは、科学者または技術者がアッセイの開発について熱プロファイルを開発および調整することを可能にする。カスタムプロファイルは、種々の加熱要件および冷却要件について開発され得る。
以下の温度プロファイルは、本発明の方法および装置を使用する微流動性カードで達成された。
1)一定性
2)アニーリング工程について調節可能で正確な温度(最低T)
3)アニーリング工程について調節可能な保持時間
4)伸長工程において調節可能な保持時間(72℃)
5)95℃を越えないこと(酵素の変性を防ぐ)
6)迅速性。
1)安定な40℃の温度(>42℃で酵素が変性される)。±1.0℃。
2)65℃および40℃での調節可能な保持時間。40℃について、最大90分間。
2)65℃以上がよい。
3)65℃において2〜5分間の保持が標準的であるが、それより短くてもよい(未知)。
4)65℃の後、40℃への一貫した時間(consitent time)(プログラムされた酵素添加のため)。
5)より短い方が良好であるが、65℃から40℃に冷却するための1〜2分間はよい。
本発明の例示された実施形態の上記の記載は、網羅的であることも、本発明を詳細な形態の開示に限定することも意図しない。当業者が理解するように、本発明の特定の実施形態、および本発明についての実施例は、例示的な目的のために本明細書中で記載されるが、種々の等価な改変は本発明の範囲内で可能である。本明細書中で提供される、本発明の教示は、他の微流動性デバイスに適用され得、必ずしも上記のPCRカードおよびrtPCRカードである必要はない。
Claims (5)
- 約15秒以下で1回の完全なポリメラーゼ連鎖反応の温度サイクルを駆動するための熱サイクリング装置であって、該装置は、以下:
a)使い捨て可能な断熱性のプラスチックカード本体であって、以下:
i)第一の表面と第二の表面とを備える第一のプレート、
ii)該第一の表面上に第一の増幅チャンバーを形成するために該第一のプレートの該第一の表面に適用される、担体と該担体の両面の接着剤層とを有する、両面接着層、
iii)該第一の増幅チャンバーに流体的に連結された微小流体アッセイ流体回路、および
iv)該第一の増幅チャンバーを覆いかつシールするために該両面接着層に接着されたプラスチック熱交換膜であって、該プラスチック熱交換膜は、該第一のプレートの該第一の表面に面した内面と、逆方向を向いた外面とを備え、該熱交換膜は、ポリエチレンテレフタレートの極薄フィルムである、プラスチック熱交換膜
を備える、プラスチックカード本体;ならびに
b)該第一の増幅チャンバーを覆う該プラスチック熱交換膜の該外面と熱的接触状態にある第一の熱移動要素
を備える、
装置。 - 請求項1に記載の熱サイクリング装置であって、ここで
前記第一の熱移動要素がさらに、約1.5W/m−°K以上の熱伝導率を有する熱パッドを備え、そして該熱パッドが該プラスチック熱交換膜の該外面と接触して配置されている、
装置。 - 約15秒以下で1回の完全なポリメラーゼ連鎖反応の温度サイクルを駆動するための熱サイクリング装置であって、以下:
a)使い捨て可能な断熱性のプラスチックカード本体であって、以下:
i)第一の表面と第二の表面とを備える第一のプレート、
ii)該第一の表面に形成される第一の増幅チャンバー、
iii)該第一の増幅チャンバーに流体的に連結された微小流体アッセイ流体回路、および
iv)該第一の増幅チャンバーを覆いかつシールするプラスチック熱交換膜であって、該プラスチック熱交換膜は、該第一のプレートの該第一の表面に面した内面と、逆方向を向いた外面とを備え、該熱交換膜は、ポリエチレンテレフタレートの極薄フィルムである、プラスチック熱交換膜
を備える、プラスチックカード本体;
b)該第一の増幅チャンバーを覆う該プラスチック熱交換膜の該外面と熱的接触状態にある第一の熱移動要素であって、該プラスチック熱交換膜の該外面と熱的接触状態にある温度制御された循環熱交換流体を備える、第一の熱移動要素;
c)溶液中の核酸ハイブリッドの変性温度以上の流体温度HotINを有する、温熱交換流体のためのポンプ;
d)溶液中の核酸ハイブリッドのアニーリング温度以下の流体温度ColdINを有する、冷熱交換流体のためのポンプ;ならびに
e)該プラスチックカード本体中で該温熱交換流体と該冷熱交換流体とを混合して、該温度制御された熱交換流体を生じるための手段であって、該手段は、以下:
i)該温熱交換流体を循環させるための第一の入口と、該冷熱交換流体を循環させるための第二の入口、
ii)該第一の入口および該第二の入口に流体的に連結された混合チャンバー、
iii)該第一の入口と該混合チャンバーとの間に置かれる温制御流体バルブと、該第二の入口と該混合チャンバーとの間に置かれる冷制御流体バルブ、および
iv)該熱交換膜の該外面を横切って該温度制御された熱交換流体を循環させるためのチャネル
を備える熱交換回路を備える、手段
を備え、
該温制御流体バルブおよび該冷制御流体バルブが、制御された量の該温熱交換流体および該冷熱交換流体を該混合チャンバーに入れ、それにより、該温度制御された熱交換流体および該増幅チャンバーの温度を制御するように構成された微小流体バルブであり、そして
該熱交換流体が、フッ素化炭化水素流体であり、該熱交換回路がさらに除泡器を備える、
装置。 - 請求項1に記載の熱サイクリング装置であって、
ここで、前記第一の熱移動要素は、該第一の増幅チャンバーを覆う該ポリエチレンテレフタレートのプラスチック熱交換膜の該外面と熱的接触状態にある熱電気冷却器である、
装置。 - 前記第一の熱移動要素がさらに、約1.5W/m−°Kよりも高い熱伝導率を有する熱パッドを備え、該熱パッドが、前記熱電気冷却器と前記プラスチック熱交換膜の前記外面との間に置かれる、請求項4に記載の熱サイクリング装置。
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Also Published As
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JP2006527369A (ja) | 2006-11-30 |
CN1812839A (zh) | 2006-08-02 |
US7544506B2 (en) | 2009-06-09 |
AU2004245094A1 (en) | 2004-12-16 |
RU2005141456A (ru) | 2006-06-10 |
KR101203402B1 (ko) | 2012-11-23 |
US20050129582A1 (en) | 2005-06-16 |
WO2004108287A1 (en) | 2004-12-16 |
KR20060017850A (ko) | 2006-02-27 |
EP1633481A1 (en) | 2006-03-15 |
CA2528208A1 (en) | 2004-12-16 |
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