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
Text Size: A A A

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.

Figures in this Article
© 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


Tables

Access This Article
Sign in or Create a personal account to Buy this article ($20 for members, $25 for non-members).

Some tools below are only available to our subscribers or users with an online account.

Related Content

Customize your page view by dragging & repositioning the boxes below.

Related Book Chapters

Topic Collections

PubMed Articles
Advertisement
  • Don't have an account?
  • Subscribe to the SPIE Digital Library
  • Create a FREE account to sign up for Digital Library content alerts and gain access to institutional subscriptions remotely.
Access This Article
Sign in or Create a personal account to Buy this article ($20 for members, $25 for non-members).
Access This Proceeding
Sign in or Create a personal account to Buy this article ($15 for members, $18 for non-members).
Access This Chapter

Access to SPIE eBooks is limited to subscribing institutions and is not available as part of a personal subscription. Print or electronic versions of individual SPIE books may be purchased via SPIE.org.