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Spectral Versatility of Fluorescent Proteins Observed on the Single Molecule Level

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Fluorescent Proteins I

Part of the book series: Springer Series on Fluorescence ((SS FLUOR,volume 11))

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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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  1. Nanobiophysics, MESA+ Institute for Nanotechnology, Faculty of Science and Technology, University of Twente, 217, 7500, AE Enschede, The Netherlands

    Christian Blum & Vinod Subramaniam

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  1. Christian Blum
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  2. Vinod Subramaniam
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Correspondence to Christian Blum .

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  1. , Campus B2 2, Saarland University, Saarbrücken, 66123, Saarland, Germany

    Gregor Jung

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Blum, C., Subramaniam, V. (2011). Spectral Versatility of Fluorescent Proteins Observed on the Single Molecule Level. In: Jung, G. (eds) Fluorescent Proteins I. Springer Series on Fluorescence, vol 11. Springer, Berlin, Heidelberg. https://doi.org/10.1007/4243_2011_22

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