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Monitoring Integrated Activity of Individual Neurons Using FRET-Based Voltage-Sensitive Dyes

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Membrane Potential Imaging in the Nervous System

Abstract

Pairs of membrane-associated molecules exhibiting fluorescence resonance energy transfer (FRET) provide a sensitive technique to measure changes in a cell’s membrane potential. One of the FRET pair binds to one surface of the membrane and the other is a mobile ion that dissolves in the lipid bilayer. The voltage-related signal can be measured as a change in the fluorescence of either the donor or acceptor molecules, but measuring their ratio provides the largest and most noise-free signal. This technology has been used in a variety of ways; three are documented in this chapter: (1) high-throughput drug screening; (2) monitoring the activity of many neurons simultaneously during a behavior; and (3) finding synaptic targets of a stimulated neuron. In addition, we provide protocols for using the dyes on both cultured neurons and leech ganglia. We also give an updated description of the mathematical basis for measuring the coherence between electrical and optical signals. Future improvements of this technique include faster and more sensitive dyes that bleach more slowly, and the expression of one of the FRET pair genetically.

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Author information

Authors and Affiliations

  1. Department of Biomedical Optics, Max Planck Institute for Medical Research, Jahnstrasse, 2969120, Heidelberg, Germany

    Kevin L Briggman

  2. Neurobiology Section, Division of Biological Sciences, University of California, San Diego 9500 Gilman Drive, La Jolla, CA, 92093-0357, USA

    William B. Kristan

  3. 6468 Wayfinders Court, Carlsbad, CA, 92011, USA

    Jesús E. González

  4. Department of Physics, University of California, San Diego, 9500 Gilman Drive, La Jolla, CA, 92093-0374, USA

    David Kleinfeld

  5. Department of Pharmacology, Howard Hughes Medical Institute, University of California, San Diego 310 George Palade Labs, 9500 Gilman Drive, La Jolla, CA, 92093-0647, USA

    Roger Y. Tsien

Authors
  1. Kevin L Briggman
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  2. William B. Kristan
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  3. Jesús E. González
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  4. David Kleinfeld
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  5. Roger Y. Tsien
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Editor information

Editors and Affiliations

  1. Université Joseph Fourier, Unité Inserm 836, Grenoble Institute of Neuroscience, Site Santé à la Tronche, BP 170, Grenoble, 38042 Gren, France

    Marco Canepari

  2. Dept. Cellular &, Molecular Physiology, Yale University School of Medicine, Cedar St. 333, New Haven, 06520, Connecticut, USA

    Dejan Zecevic

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Briggman, K.L., Kristan, W.B., González, J.E., Kleinfeld, D., Tsien, R.Y. (2010). Monitoring Integrated Activity of Individual Neurons Using FRET-Based Voltage-Sensitive Dyes. In: Canepari, M., Zecevic, D. (eds) Membrane Potential Imaging in the Nervous System. Springer, New York, NY. https://doi.org/10.1007/978-1-4419-6558-5_6

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