Spectroscopic characterization of Venus at the single molecule levelElectronic supplementary information (ESI) available. See DOI: 10.1039/c1pp05292f

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 molecu...

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Main Authors: David, Charlotte C, Dedecker, Peter, Cremer, Gert De, Verstraeten, Natalie, Kint, Cyrielle, Michiels, Jan, Hofkens, Johan
Format: Article
Language:English
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Summary: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. Time-resolved fluorescence spectroscopy on the fluorescent protein Venus reveals the presence of off-times on various time scales, which may play a crucial role in its application in single molecule experiments.
ISSN:1474-905X
1474-9092
DOI:10.1039/c1pp05292f