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One-Photon and Two-Photon Excitation of Fluorescent Proteins Read chapter Read first chapter

2012 | OriginalPaper | Chapter

Spectral Versatility of Fluorescent Proteins Observed on the Single Molecule Level

Authors : Christian Blum, Vinod Subramaniam

Published in: Fluorescent Proteins I

Publisher: Springer Berlin Heidelberg

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Abstract

The photophysics of visible fluorescent proteins (VFPs) remains a topic of intense research, driven by the widespread use of these proteins as reporters and sensors in living cells. The photophysical complexity of these markers originates from the multistep chemical reaction that forms the chromophore, and from the embedding of the chromophore within the protein nanoenvironment. To accurately interpret the biological and biochemical processes illuminated by the VFPs, it is essential to understand the details of their photophysics. Certain aspects of VFP photophysics can only be observed and understood at the single molecule level, which removes the averaging effect inherent to ensemble studies. Here, we review how spectrally resolved single molecule emission detection at room temperature has helped to understand the complex photophysics of VFPs. We focus on the detection of spectrally distinct subensembles, the spontaneous or light-induced transition between these subensembles, and on subtle spectral variations induced by changes in the local nanoenvironment of VFP chromophores.

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previous chapter Photoconversion of the Green Fluorescent Protein and Related Proteins
next chapter Structure–Function Relationships in Fluorescent Marker Proteins of the Green Fluorescent Protein Family
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Metadata
Title
Spectral Versatility of Fluorescent Proteins Observed on the Single Molecule Level
Authors
Christian Blum
Vinod Subramaniam
Copyright Year
2012
Publisher
Springer Berlin Heidelberg
Book
Fluorescent Proteins I

Print ISBN: 978-3-642-23371-5

Electronic ISBN: 978-3-642-23372-2

Copyright Year: 2012

DOI
https://doi.org/10.1007/4243_2011_22

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