Biochemical Applications Of Frequency-Domain Fluorometry; Determination Of Time-Resolved Anisotropies And Emission Spectra

Joseph R. Lakowicz; Ignazy Gryczynski; Henryh Cherek; Gabor Laczko; Nanda Joshi

doi:10.1117/12.966934

1 January 1987 Biochemical Applications Of Frequency-Domain Fluorometry; Determination Of Time-Resolved Anisotropies And Emission Spectra

Joseph R. Lakowicz, Ignazy Gryczynski, Henryh Cherek, Gabor Laczko, Nanda Joshi

Author Affiliations +

Proceedings Volume 0743, Fluorescence Detection; (1987) https://doi.org/10.1117/12.966934
Event: OE LASE'87 and EO Imaging Symposium, 1987, Los Angeles, CA, United States

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.

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Joseph R. Lakowicz, Ignazy Gryczynski, Henryh Cherek, Gabor Laczko, and Nanda Joshi "Biochemical Applications Of Frequency-Domain Fluorometry; Determination Of Time-Resolved Anisotropies And Emission Spectra", Proc. SPIE 0743, Fluorescence Detection, (1 January 1987); https://doi.org/10.1117/12.966934

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