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Enhanced yellow fluorescent protein photoconversion to a cyan fluorescent protein-like species is sensitive to thermal and diffusion conditions.
J Biomed Opt 2009 May-Jun; 14(3):034039JB

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.

Authors+Show Affiliations

Aarhus University, Stereology and Electron Microscopy Research Laboratory, Histoinformatics and MIND Centres, Ole Worms Alle 1185, DK-8000 Aarhus C., Denmark. raarup@ki.au.dkNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

19566331

Citation

Raarup, Merete K., et al. "Enhanced Yellow Fluorescent Protein Photoconversion to a Cyan Fluorescent Protein-like Species Is Sensitive to Thermal and Diffusion Conditions." Journal of Biomedical Optics, vol. 14, no. 3, 2009, p. 034039.
Raarup MK, Fjorback AW, Jensen SM, et al. Enhanced yellow fluorescent protein photoconversion to a cyan fluorescent protein-like species is sensitive to thermal and diffusion conditions. J Biomed Opt. 2009;14(3):034039.
Raarup, M. K., Fjorback, A. W., Jensen, S. M., Müller, H. K., Kjaergaard, M. M., Poulsen, H., ... Nyengaard, J. R. (2009). Enhanced yellow fluorescent protein photoconversion to a cyan fluorescent protein-like species is sensitive to thermal and diffusion conditions. Journal of Biomedical Optics, 14(3), p. 034039. doi:10.1117/1.3103338.
Raarup MK, et al. Enhanced Yellow Fluorescent Protein Photoconversion to a Cyan Fluorescent Protein-like Species Is Sensitive to Thermal and Diffusion Conditions. J Biomed Opt. 2009;14(3):034039. PubMed PMID: 19566331.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Enhanced yellow fluorescent protein photoconversion to a cyan fluorescent protein-like species is sensitive to thermal and diffusion conditions. AU - Raarup,Merete K, AU - Fjorback,Anja W, AU - Jensen,Stig M R, AU - Müller,Heidi K, AU - Kjaergaard,Maj M, AU - Poulsen,Hanne, AU - Wiborg,Ove, AU - Nyengaard,Jens R, PY - 2009/7/2/entrez PY - 2009/7/2/pubmed PY - 2009/9/23/medline SP - 034039 EP - 034039 JF - Journal of biomedical optics JO - J Biomed Opt VL - 14 IS - 3 N2 - 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. SN - 1083-3668 UR - https://www.unboundmedicine.com/medline/citation/19566331/Enhanced_yellow_fluorescent_protein_photoconversion_to_a_cyan_fluorescent_protein_like_species_is_sensitive_to_thermal_and_diffusion_conditions_ L2 - https://doi.org/10.1117/1.3103338 DB - PRIME DP - Unbound Medicine ER -