Spectroscopic characterization of Venus at the single molecule level.
Venus is a recently developed, fast maturating, yellow fluorescent protein that has been used as a probe for in vivo applications. In the present work the photophysical characteristics of Venus were analyzed spectroscopically at the bulk and single molecule level. Through time-resolved single molecule measurements we found that single molecules of Venus display pronounced fluctuations in fluorescence emission, with clear fluorescence on- and off-times. These fluorescence intermittencies were found to occupy a broad range of time scales, ranging from milliseconds to several seconds. Such long off-times can complicate the analysis of single molecule counting experiments or single-molecule FRET experiments.
Department of Chemistry, Katholieke Universiteit Leuven, Celestijnenlaan 200F, 3001, Heverlee, Belgium.
Charlotte C
CC
David
Peter
P
Dedecker
Gert
G
De Cremer
Natalie
N
Verstraeten
Cyrielle
C
Kint
Jan
J
Michiels
Johan
J
Hofkens
D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics; D004926:Escherichia coli / Q000166:cytology / Q000235:genetics; D008164:Luminescent Proteins / Q000737:chemistry* / Q000235:genetics; D057054:Molecular Imaging; D013050:Spectrometry, Fluorescence / Q000379:methods*; D013997:Time Factors
D001426:Bacterial Proteins; D008164:Luminescent Proteins; C054562:yellow fluorescent protein, Bacteria