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Quantitative microfluorometry of isolated living cells with pulsed excitation: Development of an effective and relatively inexpensive instrument

 

作者: J. Vigo,   J. M. Salmon,   P. Viallet,  

 

期刊: Review of Scientific Instruments  (AIP Available online 1987)
卷期: Volume 58, issue 8  

页码: 1433-1438

 

ISSN:0034-6748

 

年代: 1987

 

DOI:10.1063/1.1139428

 

出版商: AIP

 

数据来源: AIP

 

摘要:

Using a computer interfaced original apparatus, we can analyze the fluorescence decay time of a single living cell. This apparatus can measure rate constants in the range of one to few hundred nanoseconds. The smallest sample analyzed was 5×5×5 &mgr;m. We obtained enough sensitivity to measure the fluorescence of about 105molecules of benzo(a)pyrene with a signal/noise ratio equal to 5. The signal was then defined with 300 points and experiment was 60 s long with a laser pulse frequency equal to 100 Hz. Distortions were less than 3% of the signal. Different kinds of information became accessible: statistical comparison on cellular populations and variation with time of studied parameters. As an example of applications, the intrinsic cell fluorescence has been analyzed in order to determine the amounts of bound and free NAD(P)H in single living cells.

 

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