JP5866243B2 - コード化されたマイクロ粒子 - Google Patents
コード化されたマイクロ粒子 Download PDFInfo
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- JP5866243B2 JP5866243B2 JP2012088799A JP2012088799A JP5866243B2 JP 5866243 B2 JP5866243 B2 JP 5866243B2 JP 2012088799 A JP2012088799 A JP 2012088799A JP 2012088799 A JP2012088799 A JP 2012088799A JP 5866243 B2 JP5866243 B2 JP 5866243B2
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Description
本特許出願は、2006年1月25日出願の同時係属中の米国仮出願第60/762,238号、2005年9月13日出願の米国仮出願第60/716,694号の優先権を主張し、これらそれぞれの主題は、その全体が参照により本明細書に組み込まれる。
次に、工程134でSiO2の最上層がパターニングされたシリコン層上に堆積され、続いて、工程136で二酸化シリコン層がパターニングされて、個別の(ただしまだリリースされていない)マイクロ粒子が形成される。次に、シリコン基板をエッチングし、マイクロ粒子を個々の粒子として分離させる無指向性シリコン・エッチングによって、工程140でマイクロ粒子がシリコン基板からリリースされる。上述したような図5のフローチャートは、異なる工程におけるマイクロ粒子の断面図と平面図においてより良好に示される。断面図及び平面図は図6a〜図6mに概略的に示される。
図1aのセグメント(例えば、セグメント102)の走査型電子顕微鏡(SEM)画像が図8cに示される。図面に見られるように、セグメントの断面はほぼ正方形である。セグメントの頂部は1.0ミクロンの幅を有し、セグメントの底部の幅は1.2ミクロンである。セグメントの高さは約1ミクロンである。当然ながら、より大きな寸法又はより小さな寸法が可能である。
図9aのSEM画像における試料は、粒子の長軸に垂直にチップ分割し、次に、単に画像化のため、時間を設定してシリコンにエッチングを施して、内側のシリコンと外側の二酸化シリコンのコントラストを上げることによって、特性決定のために準備されたものである。
本発明のマイクロ粒子は、ウェハ・レベルで製造し、ウェハ・レベル又はダイ・レベルのどちらかでリリースすることができる。具体的には、一組のマイクロ粒子をそれぞれ含む複数のダイを、ウェハ上に形成することができる。各ダイ上のマイクロ粒子は同じであってもなくてもよく、すなわち、各ダイ上のマイクロ粒子は同じコードを有していてもいなくてもよい。マイクロ粒子を形成した後、ダイをウェハから分離することができ、その後、単一化されたダイ上のウェハ(1つ又は複数)を除去することができる。代表的なウェハ・レベルの製造方法が図12a〜図12cに示される。
積当たりの強度として定義され、光学画像面における視野の深さ通して積算される。この例では、空気密度は、検出器、例えばCCDカメラ又は光電子増倍管によって測定される信号強度プロファイルとして測定される。図27a〜27cは、図26a〜26cの対応する粒子に関して、粒子面に対して直角(すなわち、垂直)に測定された不均一な空気密度を示す。信号強度プロファイルは、粒子の面のへこみの場所に対応するピークを有し、それらが結果として検出可能かつ有用なコードをもたらす。図26a〜26cのコード化されたマイクロ粒子の面形体は、光散乱の測定、例えば暗視野光学顕微鏡などを包含するがそれに限定されない、発光分子を使用する以外の方法によって検出されてもよい。
、多重印刷工程はマイクロ粒子上にコードを設ける。
いことがある。
れるなど、マイクロ粒子の製造や検出において多くの利点を有する。コード体系の例では
、マイクロ粒子の製造プロセスの信頼性は、単一のサイズの機構、例えば単一の幅を有す
るセグメント内のギャップのための、パターニング及びエッチング条件を最適化できるよ
うにすることで改善される。
60倍対物レンズ及び6.2mmの1024×1024CCDチップを使用すると、d=
0.4μmの間隔距離は約4ピクセルに等しい。間隔距離が0.3μm(3ピクセル)に低減された場合、105,154個のコードとなる。より長い粒子長さL、及び/又はより小さなギャップ幅wに対して、コードスペースは数百万に拡張することができる。
別の例では、ウェハ又は基板上の全てのダイは同じコードを有してもよい。ダイのサイズ
は、コード当たりの粒子の数と大きな集合内のコードの数との間のバランスを最適化するように選択される。ダイ当たりの粒子の数及びウェハ当たりのダイの数は、例えば、コードを作成する本発明の方法を利用して、ソフトウェア内で変更され、また、固定の成形型
、例えばフォトマスクの大きく高価な集合の高い資本コストを必要とすることなく、異なる用途向けの製造ロット又は製品単位で最適化されてもよい。
画像中に示されるマイクロ粒子は、それらの面に付着されたオリゴ・プローブ分子を有し、ハイブリッド化されて、標的の塩基対配列がプローブの配列に対して相補的である、予め標識付けされた蛍光オリゴ標的となっている。
決定するために使用される。図45bの蛍光画像は、緑色光で照明し赤色光を集光して(励起フィルタ=555/28nm、放射フィルタ=617/73、すなわちCy3用フィルタ)得られたものである。図45cは、図45aと図45bの画像対を互いに重ね合わせて単一画像としたものである。
好ましい。
以下の参照文献それぞれの主題は、全体として参照により本明細書に組み込まれる。
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Claims (16)
- 軸線に沿って整列した2つ以上の別個のセグメントを含む第1の材料と、
前記セグメントが第2の材料を通して検出可能であるようにして、前記第1の材料を取り囲む第2の材料を含み、
前記2つ以上の別個のセグメントがコードを形成し、前記軸線に沿って同一の断面を有し、かつマイクロ粒子の断面が矩形であり、
前記マイクロ粒子は、前記セグメントと前記セグメントの間に挟まれたギャップを有し、その長さの異なる前記セグメントとその長さの等しい前記ギャップとが交互に整列されている、コード化されたマイクロ粒子。 - 前記第2の材料が透明である請求項1に記載のマイクロ粒子。
- 前記透明材料がガラスを含む請求項2に記載のマイクロ粒子。
- 前記マイクロ粒子の外面全体がガラスである請求項3に記載のマイクロ粒子。
- 前記マイクロ粒子の前記軸線方向に計測された長さが160μm以下である請求項1に記載のマイクロ粒子。
- 前記マイクロ粒子は、強磁性材料、反磁性材料、常磁性材料、超常磁性材料からなるグループから選択された材料を更に含む請求項1に記載のマイクロ粒子。
- 前記第2の材料がその中又は上に複数の蛍光分子を含む請求項1に記載のマイクロ粒子。
- 前記第2の材料が、多数の生化学分子がその上に付着される面を含む請求項1に記載のマイクロ粒子。
- 前記マイクロ粒子は、複数のマイクロ粒子の1つであり、前記複数のマイクロ粒子が単分子層内にほぼ配置される請求項1に記載のマイクロ粒子。
- 前記マイクロ粒子は、複数のマイクロ粒子の1つであり、前記複数のマイクロ粒子が液体中に配置され、前記マイクロ粒子がブラウン運動を行い、かつ前記面に付着された生化学分子を有する請求項8に記載のマイクロ粒子。
- 前記第2の材料がその面に複数のへこみを含む請求項1に記載のマイクロ粒子。
- 請求項1に記載された一組のマイクロ粒子を用意する工程を含み、
前記マイクロ粒子の層が分析の間コンテナの内面上に配列され、前記マイクロ粒子が前記内面上にほぼ単分子層の形で配置され、
個々のマイクロ粒子の空間コードを検出するため、前記マイクロ粒子からの電磁放射を透過させるか、又は反射させ、前記透過又は反射した電磁放射を検出する工程をさらに含む、マイクロ粒子のコードを検出する方法。 - 前記マイクロ粒子上のプローブと対応する被検体との結合を生じさせるため、前記一組のマイクロ粒子を第1の試験流体と混合する工程と、
前記マイクロ粒子を第2の洗浄流体で洗浄する工程と、
検出の間前記マイクロ粒子がその中に配置される第3の分析流体を追加する工程とを更に含む請求項12に記載のマイクロ粒子のコードを検出する方法。 - 遺伝子発現、メチル化検出、SNP遺伝子型決定法、比較ゲノムハイブリダイゼーション、マイクロRNAプロファイリング、微生物識別、免疫学的検出、抗体配列、蛋白質検査、蛋白質−蛋白質相互作用、受容体配位子アッセイ、ウィルス識別、病原体識別より構成されるグループより選択されたをアッセイを実行して、上記ステップは実施されることを特徴とする請求項13に記載のマイクロ粒子のコードを検出する方法。
- 請求項1記載の前記複数のマイクロ粒子と生物材料を有する試験試料を提供するステップと、
生化学的分子または生化学的プローブを含む前記マイクロ粒子を試験試料内の対応する生物学的被検体と結合させるステップと、
前記被検体と関連付けられた前記マイクロ粒子と前記被検体と関連付けられていない前記マイクロ粒子の空間コードを決定するステップと、
から構成され、
前記生化学的分子または前記生化学的プローブは、検出可能な標識または検出可能な標識を有する前記生化学的被検体を含むことを特徴とするバイオアッセイを実行する方法。 - コンテナは少なくとも200の容器を備え、各容器は液体バッファ内に請求項1に記載の1,000,000個以上のコード化されたマイクロ粒子を有し、各容器内の前記コード化されたマイクロ粒子は同一のコードを有し、
各容器は異なるコードに対応していることを特徴とするキット。
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US20100227279A1 (en) | 2010-09-09 |
US20120169819A1 (en) | 2012-07-05 |
JP2009516822A (ja) | 2009-04-23 |
EP2485052B1 (en) | 2015-05-06 |
EP1933817A4 (en) | 2009-11-25 |
US7745091B2 (en) | 2010-06-29 |
US20140057809A1 (en) | 2014-02-27 |
US8945811B2 (en) | 2015-02-03 |
JP5452922B2 (ja) | 2014-03-26 |
WO2007081410A3 (en) | 2009-05-07 |
US20070148599A1 (en) | 2007-06-28 |
EP2485052A1 (en) | 2012-08-08 |
US8460854B2 (en) | 2013-06-11 |
US8372576B2 (en) | 2013-02-12 |
US7745092B2 (en) | 2010-06-29 |
WO2007081410A2 (en) | 2007-07-19 |
US8088555B2 (en) | 2012-01-03 |
US20140051027A1 (en) | 2014-02-20 |
US20100297336A1 (en) | 2010-11-25 |
US20100227770A1 (en) | 2010-09-09 |
US20080038559A1 (en) | 2008-02-14 |
JP2012189600A (ja) | 2012-10-04 |
US8168368B2 (en) | 2012-05-01 |
EP1933817B1 (en) | 2014-03-12 |
US8592136B2 (en) | 2013-11-26 |
US8748079B2 (en) | 2014-06-10 |
US20120225295A1 (en) | 2012-09-06 |
US20100290018A1 (en) | 2010-11-18 |
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