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1. WO2009041032 - 蛍光検出方法

公開番号 WO/2009/041032
公開日 02.04.2009
国際出願番号 PCT/JP2008/002642
国際出願日 24.09.2008
IPC
G01N 21/64 2006.01
G物理学
01測定;試験
N材料の化学的または物理的性質の決定による材料の調査または分析
21光学的手段,すなわち,赤外線,可視光線または紫外線を使用することによる材料の調査または分析
62調査される材料が励起され,それにより光を発しまたは入射光の波長に変化を生ずるシステム
63光学的励起
64蛍光;燐光
CPC
G01N 2021/0346
GPHYSICS
01MEASURING; TESTING
NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
21Investigating or analysing materials by the use of optical means, i.e. using infra-red, visible or ultra-violet light
01Arrangements or apparatus for facilitating the optical investigation
03Cuvette constructions
0346Capillary cells; Microcells
G01N 21/05
GPHYSICS
01MEASURING; TESTING
NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
21Investigating or analysing materials by the use of optical means, i.e. using infra-red, visible or ultra-violet light
01Arrangements or apparatus for facilitating the optical investigation
03Cuvette constructions
05Flow-through cuvettes
G01N 21/6408
GPHYSICS
01MEASURING; TESTING
NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
21Investigating or analysing materials by the use of optical means, i.e. using infra-red, visible or ultra-violet light
62Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
63optically excited
64Fluorescence; Phosphorescence
6408with measurement of decay time, time resolved fluorescence
G01N 21/6428
GPHYSICS
01MEASURING; TESTING
NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
21Investigating or analysing materials by the use of optical means, i.e. using infra-red, visible or ultra-violet light
62Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
63optically excited
64Fluorescence; Phosphorescence
6428Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
G01N 21/6458
GPHYSICS
01MEASURING; TESTING
NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
21Investigating or analysing materials by the use of optical means, i.e. using infra-red, visible or ultra-violet light
62Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
63optically excited
64Fluorescence; Phosphorescence
645Specially adapted constructive features of fluorimeters
6456Spatial resolved fluorescence measurements; Imaging
6458Fluorescence microscopy
G01N 27/44726
GPHYSICS
01MEASURING; TESTING
NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
27Investigating or analysing materials by the use of electric, electro-chemical, or magnetic means
26by investigating electrochemical variables; by using electrolysis or electrophoresis
416Systems
447using electrophoresis
44704Details; Accessories
44717Arrangements for investigating the separated zones, e.g. localising zones
44721by optical means
44726using specific dyes, markers or binding molecules
出願人
  • 富士フイルム株式会社 FUJIFILM CORPORATION [JP]/[JP] (AllExceptUS)
  • 木村統安 KIMURA, Muneyasu [JP]/[JP] (UsOnly)
発明者
  • 木村統安 KIMURA, Muneyasu
代理人
  • 柳田征史 YANAGIDA, Masashi
優先権情報
2007-25540328.09.2007JP
公開言語 (言語コード) 日本語 (JA)
出願言語 (言語コード) 日本語 (JA)
指定国 (国コード)
発明の名称
(EN) FLUORESCENT DETECTION METHOD
(FR) PROCÉDÉ DE DÉTECTION PAR FLUORESCENCE
(JA) 蛍光検出方法
要約
(EN)
[PROBLEMS TO BE SOLVED] To design a fluorescent detection method for detecting fluorescence intensity of a fluorescently marked living specimen with high accuracy in a short time if fluorescence intensity of a microchip itself works as a background. [MEANS FOR SOLVING PROBLEMS] Fluorescence intensity (Ur0) of fluorescence (Lk0) generated from a microchip (11) itself equivalent to a microchip (10) having a fluorescently marked living specimen (R) supplied is detected since irradiation of excitation light (Le) starts until the fluorescence intensity (Ur0) sufficiently attenuates, and a change with time of the fluorescence intensity (Ur0) is stored. The excitation light (Le) is irradiated to the microchip (10) and in addition the living specimen (R) is supplied before the fluorescence intensity (Ur0) from the microchip (10) itself sufficiently attenuates, and the generated fluorescence intensity (Ur) is detected since the irradiation of the excitation light (Le) starts until after the living specimen (R) is supplied. A value of the change with time of the stored fluorescence intensity (Ur0) is subtracted from the fluorescence intensity (Ur) detected from the microchip (10) having the living specimen (R) supplied to detect fluorescence intensity (Uc) of fluorescence (Lk) generated from the living specimen (R).
(FR)
L'invention concerne la mise au point d'un procédé de détection par fluorescence pour détecter une intensité de fluorescence d'un spécimen vivant marqué par fluorescence, avec une précision élevée dans un temps court si l'intensité de fluorescence d'une micro-puce elle-même sert de fond. À cet effet, l'intensité de fluorescence (Ur0) d'une fluorescence (Lk0), générée à partir d'une micro-puce (11) elle-même équivalente à une micro-puce (10) ayant un spécimen vivant marqué par fluorescence (R) fourni, est détectée depuis le début de l'irradiation de lumière d'excitation (Le) jusqu'à ce que l'intensité de fluorescence (Ur0) s'atténue de manière suffisante, et un changement au cours du temps de l'intensité de fluorescence (Ur0) est stocké. La lumière d'excitation (Le) est irradiée vers la micro-puce (10) et, de plus, le spécimen vivant (R) est fourni avant que l'intensité de fluorescence (Ur0) provenant de la micro-puce (10) ne s'atténue de manière suffisante, et l'intensité de fluorescence générée (Ur) est détectée depuis le début de l'irradiation de la lumière d'excitation (Le) jusqu'au moment où le spécimen vivant (R) a été fourni. Une valeur du changement au cours du temps de l'intensité de fluorescence stockée (Ur0) est soustraite de l'intensité de fluorescence (Ur) détectée à partir de la micro-puce (10) ayant le spécimen vivant (R) fourni pour détecter une intensité de fluorescence (Uc) d'une fluorescence (Lk) générée à partir du spécimen vivant (R).
(JA)
【課題】マイクロチップ自体の蛍光強度がバックグラウンドとして作用しても、高精度かつ短時間に蛍光標識された生体試料の蛍光強度を検出する蛍光検出方法を図る。 【解決手段】蛍光標識された生体試料(R)が供給されたマイクロチップ(10)と同等のマイクロチップ(11)自体から発生する蛍光(Lk0)の蛍光強度(Ur0)を励起光(Le)の照射開始から蛍光強度(Ur0)が十分に減衰するまで検出し、蛍光強度(Ur0)の経時変化を記憶する。マイクロチップ(10)に励起光(Le)を照射すると共に、マイクロチップ(10)自体からの蛍光強度(Ur0)が十分に減衰する前に、生体試料(R)を供給し、発生する蛍光強度(Ur)を励起光(Le)の照射開始から生体試料(R)が供給された後まで検出する。生体試料(R)が供給されたマイクロチップ(10)から検出された蛍光強度(Ur)から、記憶された蛍光強度(Ur0)の経時変化の値を減算して、生体試料(R)から発生した蛍光(Lk)の蛍光強度(Uc)を検出する。
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