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Special Section on Visible Fluorescent Proteins

Energy migration alters the fluorescence lifetime of Cerulean: implications for fluorescence lifetime imaging Forster resonance energy transfer measurements

[+] Author Affiliations
Srinagesh V. Koushik, Steven S. Vogel

Laboratory of Molecular Physiology, National Institute on Alcohol Abuse and Alcoholism, Bethesda, Maryland 20852

J. Biomed. Opt. 13(3), 031204 (June 27, 2008). doi:10.1117/1.2940367
History: Received August 03, 2007; Revised October 04, 2007; Accepted October 11, 2007; Published June 27, 2008; August 08, 2008
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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 FRET efficiencies based on fluorescence lifetimes.

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© 2008 Society of Photo-Optical Instrumentation Engineers

Citation

Srinagesh V. Koushik and Steven S. Vogel
"Energy migration alters the fluorescence lifetime of Cerulean: implications for fluorescence lifetime imaging Forster resonance energy transfer measurements", J. Biomed. Opt. 13(3), 031204 (June 27, 2008). ; http://dx.doi.org/10.1117/1.2940367


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