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Journal of Biomedical Optics

Energy migration alters the fluorescence lifetime of Cerulean: implications for fluorescence lifetime imaging Forster resonance energy transfer measurements
Author(s): Srinagesh V. Koushik; Steven S. Vogel
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Paper Abstract

Förster resonance energy transfer (FRET) is a physical phenomenon used to study molecular interactions in living cells. Changes in the fluorescence lifetime of proteins genetically tagged with a donor fluorophore, such as cyan fluorescent protein or Cerulean, are used to measure energy transfer to a protein tagged with an acceptor fluorophore (yellow fluorescent protein or Venus). Increased transfer efficiency is usually interpreted as closer proximity. Resonance energy transfer is also possible between identical fluorophores. This form of FRET is called energy migration resonance energy transfer (EM-RET). Theoretically, EM-RET should not alter the lifetime or emission spectrum measured from a population of fluorophores. We find a change in the fluorescent lifetime of Cerulean that correlates with energy migration and can result in significant errors when using Cerulean as a donor to measure fluorescence lifetime imaging (FLIM)-FRET efficiencies.

Paper Details

Date Published: 1 May 2008
PDF: 9 pages
J. Biomed. Opt. 13(3) 031204 doi: 10.1117/1.2940367
Published in: Journal of Biomedical Optics Volume 13, Issue 3
Show Author Affiliations
Srinagesh V. Koushik, National Institutes of Health (United States)
Steven S. Vogel, National Institutes of Health (United States)

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