Biochemical applications of frequency-domain fluorometry; determination of time-resolved anisotropies and emission spectra

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

Research output: Contribution to journalArticle

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.

Original languageEnglish
Pages (from-to)110-116
Number of pages7
JournalProceedings of SPIE - The International Society for Optical Engineering
Volume743
DOIs
StatePublished - 1 Jan 1987

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