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Proceedings Paper

Fluorescence lifetime measurements in flow cytometry
Author(s): Wolfgang Beisker; Axel Klocke
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Paper Abstract

Fluorescence lifetime measurements provide insights int eh dynamic and structural properties of dyes and their micro- environment. The implementation of fluorescence lifetime measurements in flow cytometric systems allows to monitor large cell and particle populations with high statistical significance. In our system, a modulated laser beam is used for excitation and the phase shift of the fluorescence signal recorded with a fast computer controlled digital oscilloscope is processed digitally to determine the phase shift with respect to a reference beam by fast fourier transform. Total fluorescence intensity as well as other parameters can be determined simultaneously from the same fluorescence signal. We use the epi-illumination design to allow the use of high numerical apertures to collect as much light as possible to ensure detection of even weak fluorescence. Data storage and processing is done comparable to slit-scan flow cytometric data using data analysis system. The results are stored, displayed, combined with other parameters and analyzed as normal listmode data. In our report we discuss carefully the signal to noise ratio for analog and digital processed lifetime signals to evaluate the theoretical minimum fluorescence intensity for lifetime measurements. Applications to be presented include DNA staining, parameters of cell functions as well as different applications in non-mammalian cells such as algae.

Paper Details

Date Published: 2 May 1997
PDF: 11 pages
Proc. SPIE 2982, Optical Diagnostics of Biological Fluids and Advanced Techniques in Analytical Cytology, (2 May 1997); doi: 10.1117/12.273641
Show Author Affiliations
Wolfgang Beisker, GSF-National Research Ctr. for Environmental Health (Germany)
Axel Klocke, GSF-National Research Ctr. for Environmental Health (Germany)

Published in SPIE Proceedings Vol. 2982:
Optical Diagnostics of Biological Fluids and Advanced Techniques in Analytical Cytology
Robert C. Leif; Alexander V. Priezzhev; Toshimitsu Asakura; Robert C. Leif, Editor(s)

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