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Updated: Jun 20, 2026

How to Quantify the Fraction of Photoactivated Fluorescent Proteins in Bulk and in Live Cells
Published on: January 7, 2019
The photochemistry of fluorescent proteins: implications for their biological applications
Harriet E Seward1, Clive R Bagshaw
1Department of Biochemistry, University of Leicester, Henry Wellcome Building, Lancaster Road, Leicester, UK LE1 9HN.
Abstract:
Green fluorescent protein from Aequorea victoria, its relatives and derivatives are ubiquitous in their use as biological probes. In this tutorial review, we discuss the photochemistry of this fascinating class of proteins and illustrate some of their advantages and drawbacks in a range of applications. In particular, we focus on the ionisation states of the chromophore and how they are affected by internal and external proton transfer. Light-induced reversible and irreversible events are discussed in terms of the underlying chromophore structure. These phenomena have an influence on the interpretation of FRET (Förster resonance energy transfer), FRAP (fluorescence recovery after photobleaching), as well as single molecule studies.
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