JPH10512952A - フロー蛍光法及び装置 - Google Patents
フロー蛍光法及び装置Info
- Publication number
- JPH10512952A JPH10512952A JP8522061A JP52206196A JPH10512952A JP H10512952 A JPH10512952 A JP H10512952A JP 8522061 A JP8522061 A JP 8522061A JP 52206196 A JP52206196 A JP 52206196A JP H10512952 A JPH10512952 A JP H10512952A
- Authority
- JP
- Japan
- Prior art keywords
- photon
- excitation
- detection
- fluorescence
- particles
- 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.)
- Pending
Links
- 238000002795 fluorescence method Methods 0.000 title claims 2
- 230000005284 excitation Effects 0.000 claims abstract description 60
- 239000002245 particle Substances 0.000 claims abstract description 57
- 238000000034 method Methods 0.000 claims abstract description 41
- 238000001514 detection method Methods 0.000 claims abstract description 27
- 230000003287 optical effect Effects 0.000 claims abstract description 22
- 238000010219 correlation analysis Methods 0.000 claims description 10
- 239000007850 fluorescent dye Substances 0.000 claims description 10
- 238000001917 fluorescence detection Methods 0.000 claims description 9
- 230000000875 corresponding effect Effects 0.000 claims description 7
- 230000002596 correlated effect Effects 0.000 claims description 3
- 230000003595 spectral effect Effects 0.000 claims description 3
- 238000001228 spectrum Methods 0.000 claims 2
- 239000000463 material Substances 0.000 claims 1
- 238000000684 flow cytometry Methods 0.000 abstract description 21
- 238000005259 measurement Methods 0.000 abstract description 9
- 210000004027 cell Anatomy 0.000 description 22
- 238000009826 distribution Methods 0.000 description 15
- 238000004458 analytical method Methods 0.000 description 14
- 238000010521 absorption reaction Methods 0.000 description 7
- 238000005286 illumination Methods 0.000 description 6
- 238000000149 argon plasma sintering Methods 0.000 description 5
- 238000012921 fluorescence analysis Methods 0.000 description 5
- 239000007788 liquid Substances 0.000 description 5
- 230000035945 sensitivity Effects 0.000 description 5
- 230000007704 transition Effects 0.000 description 5
- 239000000975 dye Substances 0.000 description 4
- 238000001499 laser induced fluorescence spectroscopy Methods 0.000 description 4
- 230000001580 bacterial effect Effects 0.000 description 3
- 239000000725 suspension Substances 0.000 description 3
- 230000017531 blood circulation Effects 0.000 description 2
- 238000005314 correlation function Methods 0.000 description 2
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- 239000012530 fluid Substances 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 238000004557 single molecule detection Methods 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 108010004729 Phycoerythrin Proteins 0.000 description 1
- 208000007660 Residual Neoplasm Diseases 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 230000005856 abnormality Effects 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 230000002238 attenuated effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 239000013060 biological fluid Substances 0.000 description 1
- 229920001222 biopolymer Polymers 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000000601 blood cell Anatomy 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- -1 coumarin, rhodamine derivatives Chemical class 0.000 description 1
- 238000004163 cytometry Methods 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 230000014670 detection of bacterium Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 239000006185 dispersion Substances 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 210000003743 erythrocyte Anatomy 0.000 description 1
- 230000005281 excited state Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 230000001605 fetal effect Effects 0.000 description 1
- 238000002060 fluorescence correlation spectroscopy Methods 0.000 description 1
- 238000000166 fluorescence laser scanning microscopy Methods 0.000 description 1
- 238000005558 fluorometry Methods 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 230000005484 gravity Effects 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 238000009652 hydrodynamic focusing Methods 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 230000000984 immunochemical effect Effects 0.000 description 1
- 238000012886 linear function Methods 0.000 description 1
- 239000006194 liquid suspension Substances 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 210000004962 mammalian cell Anatomy 0.000 description 1
- 238000000386 microscopy Methods 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 238000010606 normalization Methods 0.000 description 1
- 210000003924 normoblast Anatomy 0.000 description 1
- 238000000399 optical microscopy Methods 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 230000005693 optoelectronics Effects 0.000 description 1
- 210000003463 organelle Anatomy 0.000 description 1
- 239000004033 plastic Substances 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000006862 quantum yield reaction Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 229910052594 sapphire Inorganic materials 0.000 description 1
- 239000010980 sapphire Substances 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 229910052710 silicon Inorganic materials 0.000 description 1
- 239000010703 silicon Substances 0.000 description 1
- 238000004513 sizing Methods 0.000 description 1
- 238000004611 spectroscopical analysis Methods 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Optical investigation techniques, e.g. flow cytometry
- G01N15/1434—Optical arrangements
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Optical investigation techniques, e.g. flow cytometry
- G01N15/1456—Optical investigation techniques, e.g. flow cytometry without spatial resolution of the texture or inner structure of the particle, e.g. processing of pulse signals
- G01N15/1459—Optical investigation techniques, e.g. flow cytometry without spatial resolution of the texture or inner structure of the particle, e.g. processing of pulse signals the analysis being performed on a sample stream
Landscapes
- Chemical & Material Sciences (AREA)
- Dispersion Chemistry (AREA)
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
Abstract
Description
Claims (1)
- 【特許請求の範囲】 1.蛍光粒子の励起が2光子励起で行われることを特徴とするフロー蛍光法。 2.蛍光放出が単一光子を分解することができる1以上の平行な光子カウンタ ーで検出されることを特徴とする請求項1に記載の方法。 3.前記異なる光子カウンターが前記粒子を染色するために用いられる異なる 蛍光染料に対応する異なる波長帯のために調整された適切なスペクトルフィルタ ーを用いることを特徴とする請求項2に記載の方法。 4.その流れの中のバックグラウンドカウントと本当の蛍光粒子からのカウン トとの間の識別が、時間領域における単一光子カウントの相関分析に基づくこと を特徴とする請求項1〜3のいずれか一に記載の方法。 5.前記励起及び検出が共焦光学セット・アップで行われることを特徴とする 請求項1に記載の方法。 6.回折制限焦点体積を供する2光子励起のための手段を含むことを特徴とす るフロー蛍光装置。 7.単一光子を分解することができる蛍光検出のための一又はいくつかの平行 な光子カウンターを組み込むことを特徴とする請求項6に記載の装置。 8.蛍光検出のために用いられるディテクターが粒子を染色するために用いら れる異なる蛍光染料に対応する異なる波長帯のために調整された適切なスペクト ルフィルターを用いることを特徴とする請求項7に記載の装置。 9.蛍光検出のために用いられるディテクターが高量子効率で単 一光子を分解することができるホトダイオードであることを特徴とする請求項7 に記載の装置。 10.時間領域における1以上の平行な光子ディテクターからの単一光子カウン トのバーストの分析を行う単一コリレーターを組み込むことを特徴とする請求項 6に記載の装置。 11.相関時間及び相関しきい値を含む相関パラメータが不特定の光子カウント の最適な識別のために調節可能であることを特徴とする請求項10に記載の装置。 12.前記粒子の大きさの決定のための相関光子カウントの時間の長さの決定の ための手段を含むことを特徴とする請求項10に記載の装置。 13.励起力が各々の特定の適用におけるサンプルからの最適な光子放出比率の ために調節可能であることを特徴とする請求項6に記載の装置。 14.励起及び検出のための共焦光学セット・アップを含むことを特徴とする請 求項6に記載の装置。 15.励起及び検出が共焦セット・アップで行われることを特徴とするフロー蛍 光法。 16.蛍光放出が単一光子を分解することができる1以上の平行な光子カウンタ ーで検出されることを特徴とする請求項15に記載の方法。 17.異なる光子カウンターが、粒子を染色するために用いられる異なる蛍光染 料に対応する異なる波長帯のために調整された適切なスペクトルフィルターを用 いることを特徴とする請求項15に記載の方法。 18.バックグラウンドカウントとその流れ中の本当の蛍光粒子からのカウント との間の識別が時間領域における単一光子カウントの 相関分析に基づくことを特徴とする請求項15〜17のいずれか一に記載の方法。 19.励起及び検出のための共焦光学セット・アップを含むことを特徴とするフ ロー蛍光装置。 20.単一光子を分解することができる蛍光検出のための1又はいくつかの平行 な光子カウンターを組み込むことを特徴とする請求項19に記載の装置。 21.蛍光検出のために用いられるディテクターが、前記粒子を染色するために 用いられる異なる蛍光染料に対応する異なる波長帯のために調整された適切なス ペクトルフィルターを用いることを特徴とする請求項20に記載の装置。 22.蛍光検出のために用いられるディテクターが、高い量子効率で単一光子を 分解することができるホトダイオードであることを特徴とする請求項20に記載の 装置。 23.時間領域における1又はいくつかの平行な光子ディテクターからの単一光 子カウントのバーストの分析を行う単一のコリレーターを組み込むこと特徴とす る請求項19に記載の装置。 24.相関時間及び相関しきい値を含む相関パラメータが不特定の光子カウント の最適な識別のために調節可能であることを特徴とする請求項23に記載の装置。 25.前記粒子の大きさの決定のための相関光子カウントの時間の長さの決定の ための手段を含むことを特徴とする請求項23に記載の装置。 26.励起力が、各々の特定の適用におけるサンプルからの最適な光子放出比率 のために調節可能であることを特徴とする請求項19に記載の装置。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
FI950174A FI98765C (fi) | 1995-01-16 | 1995-01-16 | Virtaussytometrinen menetelmä ja laite |
FI950174 | 1995-01-16 | ||
PCT/FI1996/000003 WO1996022521A1 (en) | 1995-01-16 | 1996-01-03 | A flow fluorometric method and device |
Related Child Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2004226683A Division JP2004347608A (ja) | 1995-01-16 | 2004-08-03 | フロー蛍光法及び装置 |
Publications (1)
Publication Number | Publication Date |
---|---|
JPH10512952A true JPH10512952A (ja) | 1998-12-08 |
Family
ID=8542323
Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP8522061A Pending JPH10512952A (ja) | 1995-01-16 | 1996-01-03 | フロー蛍光法及び装置 |
JP2004226683A Pending JP2004347608A (ja) | 1995-01-16 | 2004-08-03 | フロー蛍光法及び装置 |
Family Applications After (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2004226683A Pending JP2004347608A (ja) | 1995-01-16 | 2004-08-03 | フロー蛍光法及び装置 |
Country Status (6)
Country | Link |
---|---|
US (1) | US6177277B1 (ja) |
EP (1) | EP0804723B1 (ja) |
JP (2) | JPH10512952A (ja) |
DE (1) | DE69628542T2 (ja) |
FI (1) | FI98765C (ja) |
WO (1) | WO1996022521A1 (ja) |
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JP2002506522A (ja) * | 1997-06-09 | 2002-02-26 | グアヴァ テクノロジーズ インコーポレイテッド | 流体サンプル中のミクロ粒子を検出するための方法及び装置 |
JP2009037734A (ja) * | 2002-10-10 | 2009-02-19 | Landauer Inc | 酸化アルミニウム単結晶媒体を利用するビット方式光学データ記憶 |
WO2013121905A1 (ja) * | 2012-02-17 | 2013-08-22 | オリンパス株式会社 | 単一粒子検出を用いた光分析装置、光分析方法及び光分析用コンピュータプログラム |
JP2014503195A (ja) * | 2010-11-16 | 2014-02-13 | 1087 システムズ インコーポレイテッド | 生細胞の識別および選別用システム |
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Also Published As
Publication number | Publication date |
---|---|
FI98765B (fi) | 1997-04-30 |
FI950174A0 (fi) | 1995-01-16 |
EP0804723B1 (en) | 2003-06-04 |
US6177277B1 (en) | 2001-01-23 |
JP2004347608A (ja) | 2004-12-09 |
FI98765C (fi) | 1997-08-11 |
EP0804723A1 (en) | 1997-11-05 |
WO1996022521A1 (en) | 1996-07-25 |
FI950174A (fi) | 1996-07-17 |
DE69628542T2 (de) | 2003-12-18 |
DE69628542D1 (de) | 2003-07-10 |
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