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Fluorescent proteins for single-molecule fluorescence applications

✍ Scribed by Britta Seefeldt; Robert Kasper; Thorsten Seidel; Philip Tinnefeld; Karl-Josef Dietz; Mike Heilemann; Markus Sauer

Publisher
John Wiley and Sons
Year
2008
Tongue
English
Weight
466 KB
Volume
1
Category
Article
ISSN
1864-063X

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✦ Synopsis

Abstract

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. (© 2008 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)

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