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Quantitative intensity-based FRET approaches - a comparative snapshot

Item Type:Review
Title:Quantitative intensity-based FRET approaches - a comparative snapshot
Creators Name:Zeug, A. and Woehler, A. and Neher, E. and Ponimaskin, E.
Abstract:Foerster resonance energy transfer (FRET) has become an important tool for analyzing different aspects of interactions among biological macromolecules in their native environments. FRET analysis has also been successfully applied to study the spatiotemporal regulation of various cellular processes using genetically encoded FRET-based biosensors. A variety of procedures have been described for measuring FRET efficiency or the relative abundance of donor-acceptor complexes, based on analysis of the donor fluorescence lifetime or the spectrally resolved fluorescence intensity. The latter methods are preferable if one wants to not only quantify the apparent FRET efficiencies but also calculate donor-acceptor stoichiometry and observe fast dynamic changes in the interactions among donor and acceptor molecules in live cells. This review focuses on a comparison of the available intensity-based approaches used to measure FRET. We discuss their strengths and weaknesses in terms of FRET quantification, and provide several examples of biological applications.
Keywords:Confocal Microscopy, Fluorescence, Fluorescence Microscopy, Fluorescence Resonance Energy Transfer, Optogenetics, Animals
Source:Biophysical Journal
ISSN:0006-3495
Publisher:Biophysical Society (U.S.A.)
Volume:103
Number:9
Page Range:1821-1827
Date:7 November 2012
Official Publication:https://doi.org/10.1016/j.bpj.2012.09.031
PubMed:View item in PubMed

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