Fluorescent proteins for single-molecule fluorescence applications.
We present single-molecule fluorescence data of fluorescent proteins GFP, YFP, DsRed, and mCherry, a new derivative of DsRed. Ensemble and single-molecule fluorescence experiments proved mCherry as an ideally suited fluorophore for single-molecule applications, demonstrated by high photostability and rare fluorescence-intensity fluctuations. Although mCherry exhibits the lowest fluorescence quantum yield among the fluorescent proteins investigated, its superior photophysical characteristics suggest mCherry as an ideal alternative in single-molecule fluorescence experiments. Due to its spectral characteristics and short fluorescence lifetime of 1.46 ns, mCherry complements other existing fluorescent proteins and is recommended for tracking and localization of target molecules with high accuracy, fluorescence resonance energy transfer (FRET), fluorescence lifetime imaging microscopy (FLIM), or multicolor applications.
Seefeldt
Britta
B
Applied Laser Physics and Laser Spectroscopy, University of Bielefeld, Physics Department, Universitätsstrasse 25, 33615 Bielefeld, Germany.
Kasper
Robert
R
Seidel
Thorsten
T
Tinnefeld
Philip
P
Dietz
Karl-Josef
KJ
Heilemann
Mike
M
Sauer
Markus
M