JPWO2010021263A1 - マイクロチップ、及びマイクロチップの製造方法 - Google Patents
マイクロチップ、及びマイクロチップの製造方法 Download PDFInfo
- Publication number
- JPWO2010021263A1 JPWO2010021263A1 JP2010525662A JP2010525662A JPWO2010021263A1 JP WO2010021263 A1 JPWO2010021263 A1 JP WO2010021263A1 JP 2010525662 A JP2010525662 A JP 2010525662A JP 2010525662 A JP2010525662 A JP 2010525662A JP WO2010021263 A1 JPWO2010021263 A1 JP WO2010021263A1
- Authority
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- Japan
- Prior art keywords
- flow path
- resin film
- parallel
- microchip
- resin
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
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Abstract
Description
(たわみ角度)
まず、樹脂製フィルム10のたわみ角度について説明する。微細流路3の幅方向の断面において、微細流路3の一辺を形成している樹脂製フィルム10の下面11の任意の点における接線と、樹脂製基板2と樹脂製フィルム10との接合面12とのなす角度をたわみ角度θとして定義する。樹脂製基板2の流路用溝の壁面が接合面12に対して直交していれば、たわみ角度θの取り得る範囲は、0度以上90度以下である(0°≦θ≦90°)。なお、このたわみ角度が、流路の各位置での幅方向の断面における樹脂製フィルム10のたわみ角度に相当する。以下、流路の各位置での幅方向の断面における樹脂製フィルム10のたわみ角度を、「流路における樹脂製フィルム10のたわみ角度」と称する場合がある。
(たわみ量)
次に、樹脂製フィルム10のたわみ量tについて説明する。微細流路3の幅方向の断面において、微細流路3の一辺を形成している樹脂製フィルム10の下面11の任意点と、樹脂製基板2と樹脂製フィルム10の接合面12との距離をたわみ量として定義する。ここでの距離とは、樹脂製フィルム10の下面11の任意点から接合面12を含む平面へ向かって垂線を引いたとき、任意点から平面までの垂線の長さとする。なお、このたわみ量が、流路の各位置での幅方向の断面における樹脂製フィルム10のたわみ量に相当する。以下、流路の各位置での幅方向の断面における樹脂製フィルム10のたわみ量を、「流路における樹脂製フィルム10のたわみ量」と称する場合がある。
(たわみ角度、たわみ量の測定方法)
たわみ角度とたわみ量の測定には、オリンパス製の走査型共焦点レーザ顕微鏡OLS3000を使用した。波長λ=408nmのレーザ光源、コンフォーカル光学系、及び厳密なスキャニング機構により、高精度の測定が可能である。
(アニール処理)
樹脂製基板2と樹脂製フィルム10とを熱融着によって接合した後、マイクロチップ1をアニール処理しても良い。微細流路3と貫通孔4とにおいて樹脂製フィルム10のたわみが発生しているということは、流路用溝と貫通孔4とを覆う樹脂製フィルム10の膨張、又は加圧により、樹脂製フィルム10の厚さが減少し、結果として面積が増加した分が流路用溝や貫通孔4に押し込まれた結果ということが考えられる。樹脂製フィルム10を収縮させれば、樹脂製フィルム10のたわみを減少させることが可能となる。例えば、ガラス転移温度の前後までマイクロチップ1を加熱することで、樹脂製フィルム10が収縮し、その結果、樹脂製フィルム10のたわみが減少することができる。加熱温度及び加熱時間などのアニールの条件は、樹脂製フィルム10の物性、厚さ、流路用溝の幅、及び、貫通孔4の径によって異なるため、マイクロチップごとにアニールの条件を決めれば良い。1例として、マイクロチップ1を90℃で1時間、アニールすることで、樹脂製フィルム10のたわみを減少させることができる。
(変形例1)
次に、上記実施形態の変形例1について図5を参照して説明する。図5は、変形例1に係るマイクロチップの上面図である。
(変形例2)
次に、上記実施形態の変形例2について図6を参照して説明する。図6は、変形例2に係る樹脂製基板の上面図である。
(変形例3)
次に、上記実施形態の変形例3について図7を参照して説明する。図7は、変形例3に係る樹脂製基板の上面図である。
(実施例1)
(樹脂製基板2と樹脂製フィルム10との接合、アニール)
射出成形機で透明樹脂材料のポリメチルメタクリレート(アクリル系樹脂)(旭化成製、デルペット70NH)を成形することで、外形寸法が幅25mm×幅25mm×厚さ1mmの板状部材に、幅30μm、深さ30μmの流路用溝と、内径2mmの複数の貫通孔4とが形成された流路側の樹脂製基板を作製した。ここで、流路用溝の深さ30μmを、流路の設計値として定義する。この流路側の樹脂製基板が、上述した実施形態に係る樹脂製基板2の1例に相当する。
(接合)
流路用溝が形成された樹脂製基板2の接合面12に樹脂製フィルム10を重ねた。実施例1では、流路3Bの長さの合計Xが、流路3Aの長さの合計Yよりも長いため、流路3Bの長さ方向(X方向)と、樹脂製フィルム10のTD方向とが平行になり、流路3Aの長さ方向(Y方向)と、樹脂製フィルム10のMD方向とが平行になるように、樹脂製基板2に樹脂製フィルム10を重ねた。
(測定)
接合後、キーエンス製の走査型共焦点レーザ変位計LT9000を使用して、微細流路3における樹脂製フィルム10のたわみ量を測定した。たわみ量は形状測定モードにてフィルム最表面にレーザをフォーカスし、流路もしくは開口部(貫通孔4)と、その近傍の平面との変位から算出した。複数の箇所においてたわみ量を測定し、たわみ量の平均値を求めた。
(アニール)
次に、マイクロチップ1を90℃の恒温槽に1時間、設置することでマイクロチップ1をアニールした。アニール後において、微細流路3における樹脂製フィルム10のたわみ量を測定した。また、流路3Aの深さと流路3Bの深さとを測定し、深さの設計値(30μm)からの誤差を求めた。
(評価)
(マイクロチップを使った電気泳動テスト)
先ず、ウェル(開口部)から粘稠なポリマー溶液(ポリジメチルアクリルアミド、pDMA)を充填した。PMMAで作製された基板は親水性であるため、毛細管現象で導入することが可能であった。各ウェル(開口部)にも液面高さが等しくなるよう、ポリマー溶液を充填した。次に、塩基対数が100bpから1,000bpの蛍光標識されたDNAを試料投入用ウェル(開口部)に滴下し、直流電圧を印加して導入、分離を行った。所定の検出部にて共焦点レーザ顕微鏡を使って励起、検出を行ったところ、DNAのプラグ(バンド)は栓流を形成しながら分離される様子が観察できた。同様の電気泳動を10回行い、検出部での蛍光強度のばらつきを算出し、分析対象物(DNA)の検出の再現性を求めた。再現性は、蛍光強度のばらつきの度合いを示している。すなわち、再現性の値が小さければ、蛍光強度のばらつきが小さいことを示している。実施例1では、再現性は5%と良好な結果を示した。このように、実施例1では、蛍光強度のばらつきを小さくすることができた。
(実施例2)
実施例2では、流路3Aの長さの合計Yと、流路3Bの長さの合計Yとの比を変えた。
(評価)
実施例1と同じ条件で、電気泳動テストを行った。同様の電気泳動を10回行い、検出部での蛍光強度のばらつきを算出し、分析対象物(DNA)の検出の再現性を求めた。再現性は3%と良好な結果を示した。さらに、実施例1と比較して、検出感度が高いことを確認した。検出感度は、DNA濃度をどれくらい下げても検出できるかを指標として実験を行った。
(比較例)
次に、上記の実施例1及び実施例2に対する比較例について説明する。比較例では、樹脂製フィルム10の貼る方向を、実施例1及び実施例2に係る方向とは逆の方向にした。
(比較例1)
比較例1では、実施例1と同様に、流路3Aの長さの合計Yを40mmとし、流路3Bの長さの合計Xを60mmとした。流路3Bの長さの合計Xと流路3Aの合計Yとの比(X/Y)は、1.5となっている。
(接合)
比較例1では、流路3Bの長さの合計Xが、流路3Aの長さの合計Yよりも長い。そのため、比較例1では、流路3Bの長さ方向(X方向)と、樹脂製フィルム10のMD方向とが平行になり、流路3Aの長さ方向(Y方向)と、樹脂製フィルム10のTD方向とが平行になるように、樹脂製基板2に樹脂製フィルム10を重ねた。
(評価)
上記の実施例と同じ条件で、電気泳動テストを行った。同様の電気泳動を10回行い、検出部での蛍光強度のばらつきを算出し、分析対象物(DNA)の検出の再現性を求めた。再現性は7%になった。このように、比較例1によると、実施例1及び実施例2と比較して、再現性が悪化した。すなわち、比較例1では、蛍光強度のばらつきが大きくなった。この再現性では、分析用途によっては十分使用できる値ではあるが、医用診断などのように、高い再現性が求められる分野には適用することができない。また、検出感度も低い結果となった。
(比較例2)
比較例2では、実施例2と同様に、流路3Aの長さの合計Yを20mmとし、流路3Bの長さの合計Xを100mmとした。流路3Bの長さの合計Xと流路3Aの合計Yとの比(X/Y)は、5となっている。
(接合)
比較例2では、流路3Bの長さの合計Xが、流路3Aの長さの合計Yよりも長い。そのため、比較例2では、流路3Bの長さ方向(X方向)と、樹脂製フィルム10のMD方向とが平行になり、流路3Aの長さ方向(Y方向)と、樹脂製フィルム10のTD方向とが平行になるように、樹脂製基板2に樹脂製フィルム10を重ねた。
(評価)
上記の実施例と同じ条件で、電気泳動テストを行った。同様の電気泳動を10回行い、検出部での蛍光強度のばらつきを算出し、分析対象物(DNA)の検出の再現性を求めた。再現性は15%になった。このように、比較例2によると、実施例1及び実施例2と比較して、再現性が悪化した。この再現性では、近接する分子量のDNA標識又はタンパク質の分離が困難になるなど、実用上支障をきたすことが分かった。また、検出感度も低い結果となった。
2、20、30、40 樹脂製基板
3 微細流路
3A、3B、21、21A、21B 流路
4、22、35、45 貫通孔
10 樹脂製フィルム
11 下面
12 接合面
31、32、33、34、41、42、43、44 流路用溝
Claims (9)
- 流路用溝が形成された略四角形状の外形を有する樹脂製基板の、前記流路用溝が形成された面に樹脂製フィルムを接合することで、流路が形成されたマイクロチップであって、
前記樹脂製基板の第1の辺に平行な第1流路の長さの合計が、前記第1の辺とは直交する第2の辺に平行な第2流路の長さの合計よりも長く、
前記第1の辺と前記樹脂フィルムのTD方向とを平行にし、前記第2の辺と前記樹脂製フィルムのMD方向とを平行にして、前記樹脂製基板と前記樹脂製フィルムとが接合されたことを特徴とするマイクロチップ。 - 流路用溝が形成された樹脂製基板の、前記流路用溝が形成された面に樹脂製フィルムを接合することで、流路が形成されたマイクロチップであって、
前記流路を、前記樹脂製基板の外周に接して定義された前記樹脂製基板を囲む仮想の四角形の第1の辺に平行な成分である第1流路と、前記第1の辺とは直交する第2の辺に平行な成分である第2流路とに分けたときに、
前記第1流路の長さの合計が前記第2流路の長さの合計よりも長く、
前記第1の辺と前記樹脂製フィルムのTD方向とを平行にし、前記第2の辺と前記樹脂製フィルムのMD方向とを平行にして、前記樹脂製基板と前記樹脂製フィルムとが接合されたことを特徴とするマイクロチップ。 - 前記第1流路の長さの合計は、前記第2流路の長さの合計の2倍以上であることを特徴とする請求項1又は請求項2のいずれかに記載のマイクロチップ。
- 流路用溝が形成された略四角形状の外形を有する樹脂製基板の、前記流路用溝が形成された面に樹脂製フィルムを熱融着により接合することで、流路を有するマイクロチップを製造するマイクロチップの製造方法であって、
前記樹脂製基板の第1の辺に平行な第1流路の長さの合計が、前記第1の辺とは直交する第2の辺に平行な第2流路の長さの合計よりも長く、
前記第1の辺と前記樹脂製フィルムのTD方向とを平行にし、前記第2の辺と前記樹脂製フィルムのMD方向とを平行にして、前記樹脂製基板と前記樹脂製フィルムとを接合することを特徴とするマイクロチップの製造方法。 - 流路用溝が形成された樹脂製基板の、前記流路用溝が形成された面に樹脂製フィルムを熱融着により接合することで、流路を有するマイクロチップを製造するマイクロチップの製造方法であって、
前記樹脂製基板の外周に接して前記樹脂製基板を囲む仮想の四角形を定義し、前記流路を、前記仮想の四角形の第1の辺に平行な成分である第1流路と、前記第1の辺とは直交する第2の辺に平行な成分である第2流路とに分けたときに、
前記第1流路の長さの合計が前記第2流路の長さの合計よりも長く、
前記第1の辺と前記樹脂製フィルムのTD方向とを平行にし、前記第2の辺と前記樹脂製フィルムのMD方向とを平行にして、前記樹脂製基板と前記樹脂製フィルムとを接合することを特徴とするマイクロチップの製造方法。 - 前記第1流路の長さの合計は、前記第2流路の長さの合計の2倍以上であることを特徴とする請求項4又は請求項5のいずれかに記載のマイクロチップの製造方法。
- 前記第1流路の幅方向の断面における前記樹脂製フィルムのたわみ量が、前記第2流路の幅方向の断面における前記樹脂製フィルムのたわみ量よりも小さいことを特徴とする請求項4から請求項6のいずれかに記載のマイクロチップの製造方法。
- 前記第1流路の幅方向の断面における前記樹脂製フィルムのたわみ角度が、前記第2流路の幅方向の断面における前記樹脂製フィルムのたわみ角度よりも小さいことを特徴とする請求項4から請求項7のいずれかに記載のマイクロチップの製造方法。
- 前記接合された前記樹脂製基板と前記樹脂製フィルムとを、所定の温度でアニールすることを特徴とする請求項4から請求項8のいずれかに記載のマイクロチップの製造方法。
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