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Scanless two-photon voltage imaging.

Ruth R Sims1, Imane Bendifallah1, Christiane Grimm1

  • 1Institut de la Vision, Sorbonne Université, INSERM, CNRS, Paris, France.

Nature Communications
|June 14, 2024
PubMed
Summary
This summary is machine-generated.

Scanless two-photon voltage imaging, using parallel excitation, achieves high signal-to-noise ratio for genetically encoded voltage indicators. This advance enables simultaneous multi-cell recordings and optogenetic control in neuroscience research.

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Area of Science:

  • Neuroscience
  • Biophysics
  • Optical Imaging

Background:

  • Two-photon voltage imaging offers transformative potential for neuroscience.
  • Development of novel imaging approaches is needed for genetically encoded voltage indicators.

Purpose of the Study:

  • To demonstrate high-SNR two-photon voltage imaging using parallel excitation.
  • To characterize scanless two-photon voltage imaging with different illumination approaches and lasers.
  • To enable simultaneous optogenetic control and voltage imaging.

Main Methods:

  • Whole-cell patch-clamp electrophysiology.
  • Characterization of scanless two-photon voltage imaging with three parallel illumination approaches.
  • Utilizing lasers with varying repetition rates and wavelengths.
  • Co-expression of JEDI-2P-Kv and ChroME-ST.

Main Results:

  • High-SNR two-photon voltage imaging achieved with parallel excitation.
  • Successful voltage recordings of high-frequency spike trains and sub-threshold depolarizations.
  • Multi-cell recordings of up to fifteen neurons simultaneously using a low repetition-rate laser.
  • Simultaneous optogenetic stimulation and voltage imaging of action potentials.
  • In vivo imaging of multiple cells up to 250 µm deep in mouse barrel cortex.

Conclusions:

  • Scanless two-photon voltage imaging with parallel excitation enhances SNR for genetically encoded voltage indicators.
  • This method allows for high-resolution voltage recordings and simultaneous optogenetic manipulation.
  • The technique is applicable for in vivo multi-cell recordings in neuroscience research.