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Imaging Molecular Physiology in Cells Using FRET-Based Fluorescent Nanosensors

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Optical Fluorescence Microscopy

Abstract

The multiparameter nature of fluorescence microscopy makes it a powerful tool for the investigation of cellular function. Förster resonance energy transfer (FRET) allows the study of functional mechanisms, i.e., protein interactions, modifications, and conformational changes, at the molecular level. Of the different FRET microscopy techniques, fluorescence lifetime imaging (FLIM) provides a quantitative, robust, and sensitive read-out in living cells. The different consequences of the occurrence of FRET and the different forms of FRET bioassays are reviewed here. However, knowledge of isolated biochemical events in cells is not sufficient to understand the working of the highly interconnected cellular pathways. The expansion of the detection principle of FRET could uncover correlations between different components and events. The current focus is on the development of multipoint FRET assays that provide high-detail overviews of functional, structural, and organizational aspects of cellular machine components from which rules and causalities can be derived.

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Acknowledgments

Additional support is acknowledged from the “FLI-Cam” project in the Biophotonics program of the Federal Ministry of Science and Education (BMBF). I thank Alessandro Esposito for the figure of ubiquitinated α-synuclein.

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Authors and Affiliations

  1. Laboratory for Molecular and Cellular Systems, Department of Neuro- and Sensory Physiology, Centre for Physiology and Pathophysiology, University Medicine Göttingen, Humboldtallee 23, 37075, Göttingen, Germany

    Fred S. Wouters

  2. DFG Research Center for Molecular Physiology of the Brain and Excellence Cluster EXC171 for Microscopy on the Nanometer Scale, Göttingen, Germany

    Fred S. Wouters

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  1. Fred S. Wouters
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Correspondence to Fred S. Wouters .

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Editors and Affiliations

  1. , Nanophysics Unit, The Italian Institute of Technology - II, Via Morego 30, Genova, 16163, Italy

    Alberto Diaspro

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Wouters, F.S. (2011). Imaging Molecular Physiology in Cells Using FRET-Based Fluorescent Nanosensors. In: Diaspro, A. (eds) Optical Fluorescence Microscopy. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-15175-0_8

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