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Related Experiment Video

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In vivo Optogenetic Stimulation of the Rodent Central Nervous System
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In Vivo Optogenetics with Stimulus Calibration.

Luke T Coddington1, Joshua T Dudman2

  • 1Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA.

Methods in Molecular Biology (Clifton, N.J.)
|October 29, 2020
PubMed
Summary

Optogenetics enables precise control of neuronal activity using light. This study details methods for calibrating light stimulation in vivo using photometry and genetically encoded indicators for accurate results.

Keywords:
BehaviorDopamineFiber photometryFluorescenceGenetically encoded calcium indicatorsOptogeneticsRodentTransgenic

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Area of Science:

  • Neuroscience
  • Biotechnology
  • Optical Engineering

Background:

  • Optogenetic reagents offer precise control over neuronal activity using light, enabling both cell depolarization and hyperpolarization.
  • Controlling neuronal activity in vivo, particularly deep within the brain's scattering tissue, necessitates advanced light delivery strategies.
  • Variability in optogenetic protein expression requires methods for measuring and calibrating stimulation effects.

Purpose of the Study:

  • To describe general procedures for delivering light deep into brain tissue for optogenetic stimulation.
  • To present methods for simultaneously stimulating neurons and measuring their activity using photometry.
  • To enable precise calibration of optogenetic stimulation in vivo.

Main Methods:

  • Implantation of optical fibers for targeted light delivery to specific brain regions.
  • Utilizing viral techniques for optogenetic protein expression.
  • Employing photometry with genetically encoded activity indicators to monitor neuronal responses.
  • Simultaneous stimulation and photometry for real-time calibration.

Main Results:

  • Demonstration of a generalizable approach for in vivo optogenetic stimulation.
  • Successful simultaneous neuronal stimulation and activity monitoring.
  • Precise calibration of stimulation intensity based on measured neuronal activity.

Conclusions:

  • The described procedures facilitate accurate and reliable optogenetic control of neuronal activity in vivo.
  • Simultaneous stimulation and photometry provide a robust method for calibrating light delivery and expression levels.
  • This approach enhances the precision and reproducibility of optogenetic experiments in neuroscience research.