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Related Experiment Videos

Optical recording of action potentials with second-harmonic generation microscopy.

Daniel A Dombeck1, Mireille Blanchard-Desce, Watt W Webb

  • 1School of Applied and Engineering Physics, Cornell University, Ithaca, New York 14853, USA.

The Journal of Neuroscience : the Official Journal of the Society for Neuroscience
|January 30, 2004
PubMed
Summary

Nonlinear microscopy now optically records fast neuronal electrical activity in thick tissues. This breakthrough uses second-harmonic generation microscopy for high-resolution imaging of action potentials in Aplysia neurons.

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Area of Science:

  • Neuroscience
  • Biophysics
  • Optical Imaging

Background:

  • Nonlinear microscopy is crucial for neuroscience in thick tissues.
  • Optical recording of fast cellular electrical activity with nonlinear microscopy remains a challenge.

Purpose of the Study:

  • To develop a method for optically recording fast cellular electrical activity using nonlinear microscopy.
  • To assess the resolution, signal-to-noise ratio, and photodamage of the developed technique.

Main Methods:

  • Utilized second-harmonic generation (SHG) microscopy.
  • Employed a voltage-sensitive dye (4-[4-(dihexylamino)phenyl][ethynyl]-1-(4-sulfobutyl)pyridinium inner salt) in primary Aplysia neurons.
  • Recorded action potentials on soma and neurite membranes.

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Main Results:

  • Achieved optical recording of action potentials with 0.833 msec temporal and 0.6 micrometer spatial resolution.
  • Demonstrated a linear SHG response to membrane potential changes (~6%/100 mV).
  • Reported negligible photodamage and a signal-to-noise ratio of ~1, improvable to ~6-7 with averaging.

Conclusions:

  • Second-harmonic generation microscopy enables high-resolution optical recording of fast neuronal electrical activity.
  • This technique offers deep tissue imaging capabilities (up to ~400 microm) with minimal photodamage.
  • The method is a valuable tool for future electrophysiology studies in neuroscience.