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Two-Photon Excitation of Fluorescent Voltage-Sensitive Dyes: Monitoring Membrane Potential in the Infrared

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Book cover Membrane Potential Imaging in the Nervous System and Heart

Part of the book series: Advances in Experimental Medicine and Biology ((AEMB,volume 859))

Abstract

Functional imaging microscopy based on voltage-sensitive dyes (VSDs) has proven effective for revealing spatio-temporal patterns of activity in vivo and in vitro. Microscopy based on two-photon excitation of fluorescent VSDs offers the possibility of recording sub-millisecond membrane potential changes on micron length scales in cells that lie upwards of one millimeter below the brain’s surface. Here we describe progress in monitoring membrane voltage using two-photon excitation (TPE) of VSD fluorescence, and detail an application of this emerging technology in which action potentials were recorded in single trials from individual mammalian nerve terminals in situ. Prospects for, and limitations of this method are reviewed.

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Acknowledgments

Supported by USPHS grants NS40966 and NS16824 (B.M.S.) and by a Fellowship from Oak Ridge Institute for Science and Education (J.A.N.F.).

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Authors and Affiliations

  1. New York Medical College, Valhalla, NY, USA

    Jonathan A. N. Fisher

  2. University of Pennsylvania School of Medicine, Philadelphia, PA, USA

    Brian M. Salzberg

  3. Department of Neuroscience, Perelman School of Medicine, University of Pennsylvania, 418 Guardian Drive, Philadelphia, PA, 19104-6074, USA

    Brian M. Salzberg

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  1. Jonathan A. N. Fisher
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Correspondence to Brian M. Salzberg .

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Editors and Affiliations

  1. Laboratoire Interdisciplinaire de Physique, Grenoble, France

    Marco Canepari

  2. Department of Cellular and Molecular Physiology, Yale University School of Medicine, New Haven, Connecticut, USA

    Dejan Zecevic

  3. L’Institut de rythmologie et mode´lisation cardiaque LIRYC Universite´ de Bordeaux, Bordeaux, France

    Olivier Bernus

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Fisher, J.A.N., Salzberg, B.M. (2015). Two-Photon Excitation of Fluorescent Voltage-Sensitive Dyes: Monitoring Membrane Potential in the Infrared. In: Canepari, M., Zecevic, D., Bernus, O. (eds) Membrane Potential Imaging in the Nervous System and Heart. Advances in Experimental Medicine and Biology, vol 859. Springer, Cham. https://doi.org/10.1007/978-3-319-17641-3_17

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