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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