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Journal of Biomedical Optics

Enhanced yellow fluorescent protein photoconversion to a cyan fluorescent protein-like species is sensitive to thermal and diffusion conditions
Author(s): Merete Krog Raarup; Anja W. Fjorback; Stig M. R. Jensen; Heidi K. Muller; Maj M. Kjaergaard; Hanne Poulsen; Ove Wiborg; Jens R. Nyengaard
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Paper Abstract

Ongoing research efforts into fluorescent proteins continuously generates new mutation variants, some of which can become photoactivated or photoconverted to a red-shifted color upon intense UV or blue light illumination. We report a built-in propensity for enhanced yellow fluorescent protein (EYFP) to undergo irreversible photoconversion into a cyan fluorescent protein (CFP)-like species upon green-light illumination. The photoconversion is thermally activated, happens mainly in fixed, nonsealed cell samples, and may result in a very bright and relatively photostable CFP-like species. The photoconversion efficiency depends on the sample diffusivity and is much increased in dehydrated, oxygenated samples. Given the large variations in conversion efficiency observed among samples as well as within a sample, photoconversion cannot be appropriately accounted for in the analysis of acceptor photobleaching fluorescence resonance energy transfer (pbFRET) images and should rather be completely avoided. Thus, samples should always be checked and discarded if photoconversion is observed.

Paper Details

Date Published: 1 May 2009
PDF: 12 pages
J. Biomed. Opt. 14(3) 034039 doi: 10.1117/1.3103338
Published in: Journal of Biomedical Optics Volume 14, Issue 3
Show Author Affiliations
Merete Krog Raarup, Aarhus Univ. (Denmark)
Anja W. Fjorback, Aarhus Univ. (Denmark)
Stig M. R. Jensen, Aarhus Univ. (Denmark)
Heidi K. Muller, Aarhus Univ. (Denmark)
Maj M. Kjaergaard, Aarhus Univ. (Denmark)
Hanne Poulsen, Aarhus Univ. (Denmark)
Ove Wiborg, Aarhus Univ. (Denmark)
Jens R. Nyengaard, Aarhus Univ. (Denmark)

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