Time Resolved Fluorescence Spectroscopy and Diffusion of Biological Molecules
The use of lasers in time resolved fluorescence measurements leads to considerable improvement of existing techniques for the measurement of rotational diffusion and has opened new ways for the measurement of translational diffusion. The high sensitivity and spectroscopic selectivity of fluorescence labels have proven to be of great value and the diffusion of molecules can be followed at very low concentrations and in the presence of other non labelled molecules. These features are very important for the analysis of macromolecular assemblies such as enzymes or structural proteins interacting with nucleic acids of high molecular weight or membranes and their constituents. The high time resolution covering the picosecond domain has already made feasible studies of fast intramolecular motions of biomolecular structures (Munro et al.1979). An outline of concepts and methods for the determination of rotational and translational diffusion from the analysis of time dependent fluorescence anisotropy, fluorescence intensity fluctuation and fluorescence photobleaching recovery together with applications will be given.
KeywordsRotational Diffusion Translational Diffusion Time Resolve Fluorescence Spectroscopy Rotational Relaxation Time Swedish Natural Science Research Council
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