Open AccessInfluence of cell growth conditions and medium composition on EGFP photostability in live cells
Author(s) -
Anastasia V. Mamontova,
Alexey M. Bogdanov,
Konstantin A. Lukyanov
Publication year - 2015
Publication title -
biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/000114289
Subject(s) - green fluorescent protein , photobleaching , fluorescence , fluorescence recovery after photobleaching , biophysics , fluorescence microscope , chemistry , cell culture , cell growth , cell , biochemistry , growth medium , microbiology and biotechnology , biology , bacteria , genetics , gene , physics , quantum mechanics , membrane
Photostability is a key characteristic of fluorescent proteins. It was recently demonstrated that green fluorescent protein (GFP) photobleaching in live cells can be suppressed by changes in medium composition. Here we show that Ham's F12 medium provides very high enhanced GFP (EGFP) photostability during fluorescence microscopy of live cells. This property of Ham's F12 medium is associated with decreased concentrations of riboflavin and pyridoxine, and increased concentrations of FeSO 4 , cyanocobalamine, lipoic acid, hypoxanthine, and thymidine compared with DMEM. We also found that the rate of EGFP photobleaching strongly depends on cell growth conditions such as cell density and the concentration of serum. We conclude that both imaging medium composition and the physiological state of the cells can strongly affect the photostability of fluorescent proteins. Thus, accurate comparison of the photostabilities of fluorescent proteins should be performed only in side-by-side analysis in identical cell growth conditions and media.
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