Fluorescence lifetime as a new parameter in analytical cytology measurements

John A. Steinkamp; Chiranjit Deka; Bruce E. Lehnert; Harry A. Crissman

doi:10.1117/12.239510

10 May 1996 Fluorescence lifetime as a new parameter in analytical cytology measurements

John A. Steinkamp, Chiranjit Deka, Bruce E. Lehnert, Harry A. Crissman

Proceedings Volume 2678, Optical Diagnostics of Living Cells and Biofluids; (1996) https://doi.org/10.1117/12.239510
Event: Photonics West '96, 1996, San Jose, CA, United States

Abstract

A phase-sensitive flow cytometer has been developed to quantify fluorescence decay lifetimes on fluorochrome-labeled cells/particles. This instrument combines flow cytometry (FCM) and frequency-domain fluorescence spectroscopy measurement principles to provide unique capabilities for making phase-resolved lifetime measurements, while preserving conventional FCM capabilities. Cells are analyzed as they intersect a high-frequency, intensity-modulated (sine wave) laser excitation beam. Fluorescence signals are processed by conventional and phase-sensitive signal detection electronics and displayed as frequency distribution histograms. In this study we describe results of fluorescence intensity and lifetime measurements on fluorescently labeled particles, cells, and chromosomes. Examples of measurements on intrinsic cellular autofluorescence, cells labeled with immunofluorescence markers for cell-surface antigens, mitochondria stains, and on cellular DNA and protein binding fluorochromes will be presented to illustrate unique differences in measured lifetimes and changes caused by fluorescence quenching. This innovative technology will be used to probe fluorochrome/molecular interactions in the microenvironment of cells/chromosomes as a new parameter and thus expand the researchers' understanding of biochemical processes and structural features at the cellular and molecular level.

Citation Download Citation

John A. Steinkamp, Chiranjit Deka, Bruce E. Lehnert, and Harry A. Crissman "Fluorescence lifetime as a new parameter in analytical cytology measurements", Proc. SPIE 2678, Optical Diagnostics of Living Cells and Biofluids, (10 May 1996); https://doi.org/10.1117/12.239510

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