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TitleBlue-Shifted Green Fluorescent Protein Homologues Are Brighter than Enhanced Green Fluorescent Protein under Two-Photon Excitation.
Publication TypeJournal Article
Year of Publication2017
AuthorsMolina, Rosana S., Tam M. Tran, Robert E. Campbell, Gerard G. Lambert, Anya Salih, Nathan C. Shaner, Thomas E. Hughes, and Mikhail Drobizhev
JournalJ Phys Chem Lett
Volume8
Issue12
Pagination2548-2554
Date Published2017 Jun 15
ISSN1948-7185
KeywordsColor, Fluorescent Dyes, Green Fluorescent Proteins, Hot Temperature, Luminescent Proteins, Models, Molecular, Photons, Spectrometry, Fluorescence
Abstract

Fluorescent proteins (FPs) are indispensable markers for two-photon imaging of live tissue, especially in the brains of small model organisms. The quantity of physiologically relevant data collected, however, is limited by heat-induced damage of the tissue due to the high intensities of the excitation laser. We seek to minimize this damage by developing FPs with improved brightness. Among FPs with the same chromophore structure, the spectral properties can vary widely due to differences in the local protein environment. Using a physical model that describes the spectra of FPs containing the anionic green FP (GFP) chromophore, we predict that those that are blue-shifted in one-photon absorption will have stronger peak two-photon absorption cross sections. Following this prediction, we present 12 blue-shifted GFP homologues and demonstrate that they are up to 2.5 times brighter than the commonly used enhanced GFP (EGFP).

DOI10.1021/acs.jpclett.7b00960
Alternate JournalJ Phys Chem Lett
PubMed ID28530831
PubMed Central IDPMC5474692
Grant ListR01 GM109984 / GM / NIGMS NIH HHS / United States
U01 NS094246 / NS / NINDS NIH HHS / United States