Bright cyan fluorescent protein variants identified by fluorescence lifetime screening.
Optimization of autofluorescent proteins by intensity-based screening of bacteria does not necessarily identify the brightest variant for eukaryotes. We report a strategy to screen excited state lifetimes, which identified cyan fluorescent proteins with long fluorescence lifetimes (>3.7 ns) and high quantum yields (>0.8). One variant, mTurquoise, was 1.5-fold brighter than mCerulean in mammalian cells and decayed mono-exponentially, making it an excellent fluorescence resonance energy transfer (FRET) donor.
Goedhart
Joachim
J
Swammerdam Institute for Life Sciences, Section of Molecular Cytology, Centre for Advanced Microscopy, University of Amsterdam, Amsterdam, The Netherlands.
van Weeren
Laura
L
Hink
Mark A
MA
Vischer
Norbert O E
NO
Jalink
Kees
K
Gadella
Theodorus W J
TW
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000032:analysis / Q000737:chemistry* / Q000145:classification*; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000737:chemistry* / Q000145:classification*; D008856:Microscopy, Fluorescence / Q000379:methods*; D008969:Molecular Sequence Data; D020539:Sequence Analysis, Protein / Q000379:methods*
D001426:Bacterial Proteins; C485182:Cyan Fluorescent Protein; D049452:Green Fluorescent Proteins