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Proceedings Paper

Biochemical Applications Of Frequency-Domain Fluorometry; Determination Of Time-Resolved Anisotropies And Emission Spectra
Author(s): Joseph R. Lakowicz; Ignazy Gryczynski; Henryh Cherek; Gabor Laczko; Nanda Joshi
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Paper Abstract

Measurements of time-resolved fluorescence are often used for studies of biological macromolecules. Such measurements are usually performed in the time-domain, by measurement of the time-dependent emission following pulsed excitation. It has recently become possible to measure the frequency-response of the emission to intensity modulated light, over a wide range of modulation frequencies. We used frequency-domain fluorometers which operates from 1 to 220 MHz, and more recently to 2000 MHz. The frequency-domain data provide excellent resolution of time-dependent spectral parameters. It is now possible to resolve closely spaced fluorescence lifetimes, to determine multi-exponential decays of anisotropy and to determine time-resolved emission spectra of samples which display time-dependent spectral shifts. In this article we show representative results on tryptophan fluorescence from proteins and for protein-bound fluorophores.

Paper Details

Date Published: 1 January 1987
PDF: 7 pages
Proc. SPIE 0743, Fluorescence Detection, (1 January 1987); doi: 10.1117/12.966934
Show Author Affiliations
Joseph R. Lakowicz, University of Maryland (United States)
Ignazy Gryczynski, University of Maryland (United States)
Henryh Cherek, University of Maryland (United States)
Gabor Laczko, University of Maryland (United States)
Nanda Joshi, University of Maryland (United States)


Published in SPIE Proceedings Vol. 0743:
Fluorescence Detection
E. Roland Menzel, Editor(s)

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