Monitoring Membrane Voltage Using Two-Photon Excitation of Fluorescent Voltage-Sensitive Dyes

  • Jonathan A. N. Fisher
  • Brian M. Salzberg


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


Optical Recording Intrinsic Optical Signal Nonlinear Optical Phenomenon High Excitation Intensity Excitation Laser Beam 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



Supported by USPHS grants NS40966 and NS16824 (B.M.S.) and by a Bristol-Myers Squibb Postdoctoral Fellowship in Basic Neuro-sciences at The Rockefeller University (J.A.N.F.)


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Authors and Affiliations

  • Jonathan A. N. Fisher
    • 1
  • Brian M. Salzberg
    • 2
  1. 1.The Rockefeller UniversityNew York CityUSA
  2. 2.Department of NeuroscienceUniversity of Pennsylvania School of MedicinePhiladelphiaUSA

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