1 May 2008 Energy migration alters the fluorescence lifetime of Cerulean: implications for fluorescence lifetime imaging Forster resonance energy transfer measurements
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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.
© (2008) Society of Photo-Optical Instrumentation Engineers (SPIE)
Srinagesh V. Koushik, Srinagesh V. Koushik, Steven S. Vogel, Steven S. Vogel, } "Energy migration alters the fluorescence lifetime of Cerulean: implications for fluorescence lifetime imaging Forster resonance energy transfer measurements," Journal of Biomedical Optics 13(3), 031204 (1 May 2008). https://doi.org/10.1117/1.2940367 . Submission:
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