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Journal of Biomedical Optics

Tracing apoptosis and stimulation in individual cells by fluorescence intensity and anisotropy decay
Author(s): Dror Fixler; Reuven Tirosh; Naomi Zurgil; Mordechai Deutsch
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Paper Abstract

Presented is the use of fluorescence lifetime (FLT), anisotropy decay, and associated parameters as differential indicators of cellular activity. A specially designed combination of a frequency mode based time resolved microscope and a picoliter well-per-cell array have been used to perform temporal measurements in individual cells under various biological conditions. Two biological models have been examined: mitogenic activation of peripheral blood mononuclear cells (PBMC) and induction of programmed cell death (apoptosis) in Jurkat T cells (JTC). The FLT of fluorescein stained PBMC was found to increase from 4±0.02 to 4.5±0.025 ns due to mitogenic activation, whereas during apoptosis in fluorescein stained JTC, the FLT remained constant. Notably, the rotational correlation times changed in both models: decreased in PBMC from 2.5±0.08 to 2±0.1 ns, and increased in JTC from 2.1±0.07 to 3.3±0.09 ns. FLT and rotational correlation time were used to calculate the steady state fluorescence anisotropy (FA) which was compared to directly measured FA values. The present study suggests that in addition to bioindication, the said parameters can provide valuable information about cellular mechanisms that may involve complex molecular diffusion dynamics, as well as information about structural changes that a cellular fluorophore undergoes in the course of cell activation.

Paper Details

Date Published: 1 May 2005
PDF: 8 pages
J. Biomed. Opt. 10(3) 034007 doi: 10.1117/1.1924712
Published in: Journal of Biomedical Optics Volume 10, Issue 3
Show Author Affiliations
Dror Fixler, Bar-Ilan Univ. (Israel)
Reuven Tirosh, Bar-Ilan Univ. (Israel)
Naomi Zurgil, Bar-Ilan Univ. (Israel)
Mordechai Deutsch, Bar-Ilan Univ. (Israel)

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