JP5055292B2 - 共焦点イメージング方法および装置 - Google Patents
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- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
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- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
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- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
- G01N21/6452—Individual samples arranged in a regular 2D-array, e.g. multiwell plates
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- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
- G01N21/6456—Spatial resolved fluorescence measurements; Imaging
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- G02B21/0036—Scanning details, e.g. scanning stages
- G02B21/004—Scanning details, e.g. scanning stages fixed arrays, e.g. switchable aperture arrays
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N2021/6417—Spectrofluorimetric devices
- G01N2021/6421—Measuring at two or more wavelengths
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Description
本発明は、このようなニーズに応えるためのマイクロアレイイメージングおよび解析に対する新規な方法を提供するものである。この手法は、マイクロビーズからなるアレイ、フォトリソグラフィ、プリンティング技法、電気化学などをはじめとして、広範囲にわたるマイクロアレイ技術で利用できる。この手法では、マイクロアレイを共焦点ライン走査して基板上の個別対象部位を画像化することに頼っている。走査されるラインには、マイクロアレイ上の対象部位の順に並んだラインに向かって共焦点的に導かれる、複数のレーザからの合成波長による励起で生じるレトロビームに含まれる、異なる色を読み取るために、一対の補色主波長など、光の2つ以上の波長が含まれることがある。共焦点ライン走査を用いることで、アレイの隣り合った対象部位の不要な励起が原因で生じるクロストークの可能性を著しく低減しつつ、マイクロアレイのイメージング速度が大幅に改善される。
RR=((1.22)(λ)(f))/D
式中、λは波長、fは焦点距離、Dは検出対象となる2つの物体間の距離である。この用語は、たとえば、ヘクト(Hecht)、オプティクス(Optics)、第4版、アディソン・ウィズレー(Addison Wesley)、ボストン、マサチューセッツ州(2001年)(本願明細書に参照により援用する)に記載されているようなオプティクスの当該技術分野における使用法と一致することを意図している。
Claims (15)
- (a)少なくとも試料の一部領域に励起放射線を送るよう配置された放射線源と、
(b)矩形の検出器アレイと、
(c)前記試料の一部領域の矩形画像を前記矩形の検出器アレイまで導くよう配置されたイメージングオプティクスと、
(d)前記試料をスキャン軸方向に走査するよう構成されて、前記走査によって前記矩形の検出器アレイで矩形画像を形成する前記試料の一部領域が入れ替わって走査される走査デバイスと、が設けられたイメージング装置であって、
前記矩形の検出器アレイの2つの寸法方向のうち短い方、および前記画像の2つの寸法方向のうち短い方が、前記スキャン軸方向であり、
前記矩形の検出器アレイが、前記短い方とそれに直交する方にそれぞれ複数の画素を有する2次元の検出器アレイであり、
前記矩形の検出器アレイの2つの寸法方向のうち前記短い方が、共焦点性の得られる単軸であり、該単軸に共焦点性の得られるに十分な短さとなるように、前記矩形の検出器アレイの前記短い方の前記複数の画素のうち、所定の画素数が選択されている、
ことを特徴とするイメージング装置。 - 前記放射線源からの励起放射線を受けて一定方向に伝搬される放射線ラインを前記試料に送るよう位置決めされたライン生成器をさらに含むことを特徴とする請求項1に記載のイメージング装置。
- 前記放射線ラインの2つの寸法方向のうちの短い方が、前記矩形の検出器アレイの単軸において共焦点性を得られるだけの短さであり、前記単軸が、前記矩形の検出器アレイの2つの寸法方向のうち前記短い方であることを特徴とする請求項2に記載のイメージング装置。
- 並進ステージに支持されるマイクロアレイをさらに含み、これによって前記矩形の検出器アレイが前記試料に供給されていることを特徴とする請求項1に記載のイメージング装置。
- 前記矩形の検出器アレイが、垂直軸における画素素子が単一の値として読み出されるTDI(時間遅延積分)動作用に構成されたことを特徴とする請求項1に記載のイメージング装置。
- 前記矩形の検出器アレイの、2つの寸法方向における検出素子数の比であるアスペクト比が20よりも大きいことを特徴とする請求項1に記載のイメージング装置。
- 前記試料の画像が、波長、焦点距離、検出対象となる2つの物体間の距離の関数であるレイリー分解能において0.2から10マイクロメートルであることを特徴とする請求項1に記載のイメージング装置。
- 試料の画像を得る方法であって、
(a)少なくとも試料の第1の部分を、前記試料の第1の部分から放射線が発せられる条件下で励起放射線と接触させるステップと、
(b)前記試料の第1の部分から発せられている前記放射線を矩形の検出器アレイに導いて、該矩形の検出器アレイで前記試料の第1の部分の矩形画像を形成するステップと、
(c)前記試料をスキャン軸方向に走査するステップであって、ステップ(a)と(b)とを繰り返し、前記矩形の検出器アレイで前記試料の第2の部分の矩形画像を形成するステップと、を含み、
前記矩形の検出器アレイの2つの寸法方向のうち短い方と、前記画像の2つの寸法方向のうち短い方を、前記スキャン軸方向とし、
前記矩形の検出器アレイを前記短い方とそれに直交する方にそれぞれ複数の画素を有する2次元の検出器アレイとし、
前記矩形の検出器アレイの2つの寸法方向のうち前記短い方を、共焦点性を得る単軸とすると共に、該単軸に共焦点性を得るに十分な短さとするように、前記短い方の前記複数の画素のうち、所定の画素数を選択する
ことを特徴とする試料の画像を得る方法。 - 前記少なくとも試料の一部と接触する前記励起放射線が一定方向に伝搬される放射線ラインを含むことを特徴とする請求項8に記載の試料の画像を得る方法。
- 前記放射線ラインの2つの寸法方向のうちの短い方が、前記矩形の検出器アレイの単軸において共焦点性を実現できるだけの短さとし、前記単軸が、前記矩形の検出器アレイの2つの寸法方向のうち前記短い方とすることを特徴とする請求項9に記載の試料の画像を得る方法。
- 前記試料を走査するステップが、前記試料を動かすことによって前記矩形画像と前記矩形の検出器アレイとの相対位置を前記スキャン軸方向に変化させるステップを含むことを特徴とする請求項8に記載の試料の画像を得る方法。
- 前記走査するステップが垂直軸における画素素子が単一の値として読み出されるTDI(時間遅延積分)を含むことを特徴とする請求項8に記載の試料の画像を得る方法。
- 前記試料には複数の個別対象部位を有するマイクロアレイが含まれることを特徴とする請求項8に記載の試料の画像を得る方法。
- 0.1から50マイクロメートルの範囲の距離だけ離れている個別対象部位同士を区別することをさらに含むことを特徴とする請求項8に記載の試料の画像を得る方法。
- 前記試料の前記画像が波長、焦点距離、検出対象となる2つの物体間の距離の関数であるレイリー分解能において0.2から10マイクロメートルとされることを特徴とする請求項8に記載の試料の画像を得る方法。
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US11/286,309 US7329860B2 (en) | 2005-11-23 | 2005-11-23 | Confocal imaging methods and apparatus |
PCT/US2006/045058 WO2007062039A2 (en) | 2005-11-23 | 2006-11-21 | Confocal imaging methods and apparatus |
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US (6) | US7329860B2 (ja) |
EP (2) | EP2594981B1 (ja) |
JP (1) | JP5055292B2 (ja) |
CN (1) | CN101361015B (ja) |
CA (1) | CA2632221C (ja) |
DK (2) | DK1955102T3 (ja) |
ES (2) | ES2635094T3 (ja) |
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KR102645654B1 (ko) * | 2021-12-22 | 2024-03-13 | (주)얼라인드제네틱스 | 고효율 다중 스펙트럼 검출을 위한 라인 공초점 세포 계수기 |
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CN101361015A (zh) | 2009-02-04 |
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WO2007062039A2 (en) | 2007-05-31 |
US7329860B2 (en) | 2008-02-12 |
CA2632221C (en) | 2013-08-27 |
DK1955102T3 (da) | 2013-05-13 |
JP2009517662A (ja) | 2009-04-30 |
EP2594981A2 (en) | 2013-05-22 |
US8158926B2 (en) | 2012-04-17 |
US20110204212A1 (en) | 2011-08-25 |
US7960685B2 (en) | 2011-06-14 |
CA2632221A1 (en) | 2007-05-31 |
CN101361015B (zh) | 2012-11-21 |
US20100012825A1 (en) | 2010-01-21 |
EP1955102B1 (en) | 2013-04-24 |
EP1955102A2 (en) | 2008-08-13 |
WO2007062039A3 (en) | 2007-07-19 |
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