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Journal of Biomedical Optics

Characterization of an improved donor fluorescent protein for Förster resonance energy transfer microscopy
Author(s): Richard N. Day; Cynthia F. Booker; Ammasi Periasamy
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Paper Abstract

The genetically encoded fluorescent proteins (FP), used in combination with Förster resonance energy transfer (FRET) microscopy, provide the tools necessary for the direct visualization of protein interactions inside living cells. Typically, the Cerulean and Venus variants of the cyan and yellow FPs are used for FRET studies, but there are limitations to their use. Here, Cerulean and the newly developed monomeric Teal FP (mTFP) are compared as FRET donors for Venus using spectral and fluorescence lifetime measurements from living cells. The results demonstrate that when compared to Cerulean, mTFP has increased brightness, optimal excitation using the standard 458-nm laser line, increased photostability, and improved spectral overlap with Venus. In addition, the two-photon excitation and fluorescence lifetime characteristics are determined for mTFP. Together, these measurements indicate that mTFP is an excellent donor fluorophore for FRET studies, and that its use may improve the detection of interactions involving proteins that are difficult to express, or that need to be produced at low levels in cells.

Paper Details

Date Published: 1 May 2008
PDF: 9 pages
J. Biomed. Opt. 13(3) 031203 doi: 10.1117/1.2939094
Published in: Journal of Biomedical Optics Volume 13, Issue 3
Show Author Affiliations
Richard N. Day, Univ. of Virginia Health System (United States)
Cynthia F. Booker, Univ. of Virginia (United States)
Ammasi Periasamy, Univ. of Virginia (United States)


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