Abstract
Optogenetic reagents allow for depolarization and hyperpolarization of cells with light. This provides unprecedented spatial and temporal resolution to the control of neuronal activity both in vitro and in vivo. In the intact animal this requires strategies to deliver light deep into the highly scattering tissue of the brain. A general approach that we describe here is to implant optical fibers just above brain regions targeted for light delivery. In part due to the fact that expression of optogenetic proteins is accomplished by techniques with inherent variability (e.g., viral expression levels), it also requires strategies to measure and calibrate the effect of stimulation. Here we describe general procedures that allow one to simultaneously stimulate neurons and use photometry with genetically encoded activity indicators to precisely calibrate stimulation.
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Acknowledgments
JTD is a senior group leader at the Janelia Research Campus of the Howard Hughes Medical Institute (HHMI). The work described here was funded by HHMI.
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Coddington, L.T., Dudman, J.T. (2021). In Vivo Optogenetics with Stimulus Calibration. In: Dallas, M., Bell, D. (eds) Patch Clamp Electrophysiology. Methods in Molecular Biology, vol 2188. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0818-0_14
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DOI: https://doi.org/10.1007/978-1-0716-0818-0_14
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