Understanding brain function requires technologies that can control the activity of large populations of neurons with high fidelity in space and time. We developed a multiphoton holographic approach to activate or suppress the activity of ensembles of cortical neurons with cellular resolution and sub-millisecond precision. Since existing opsins were inadequate, we engineered new soma-targeted (ST) optogenetic tools, ST-ChroME and IRES-ST-eGtACR1, optimized for multiphoton activation and suppression. Employing a three-dimensional all-optical read–write interface, we demonstrate the ability to simultaneously photostimulate up to 50 neurons distributed in three dimensions in a 550 × 550 × 100-µm3 volume of brain tissue. This approach allows the synthesis and editing of complex neural activity patterns needed to gain insight into the principles of neural codes.
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We thank M. Feller, A. Naka, and J. Brown for critical feedback on the manuscript and discussions. We thank C. Baker and M. Bolton for soma-targeted ChR2 AAVs. We thank M. Li and the UC Berkeley Vision Science Core, Gene Delivery Module, for preparation of AAVs (supported by NIH Core Grant P30 EY003176). We deeply appreciate the efforts of D. Chu, C. Douglas, and R. Hakim for important technical assistance. We thank D. Taylor for help with mouse work and histology. H.A. is a New York Stem Cell Foundation-Robertson Investigator. This work was supported by The New York Stem Cell Foundation and by grants from the Arnold and Mabel Beckman Foundation, NINDS grant DP2NS087725-01, the McKnight Foundation, NINDS award F32NS095690-01 to A.R.M., the Simon’s Foundation Collaboration for the Global Brain award 415569 to I.A.O., and a fellowship from the David and Lucille Packard Foundation to L.W. This research was developed with funding from the Defense Advanced Research Projects Agency (DARPA), Contract No. N660011-17-C-4015.
The authors declare no competing interests.
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Supplementary Figures 1–23
This table provides details on each optical stimulation path used in this study and provides a quick reference to determine which experiments were performed using each setup
This table provides technical details and a calculation tool to allow readers to set up a 3D-SHOT stimulation path
This video demonstrates high speed switching of holograms at 300 Hz
Average dF/F movies of ensembles stimulation data at 30 Hz as in Fig7-8
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Mardinly, A.R., Oldenburg, I.A., Pégard, N.C. et al. Precise multimodal optical control of neural ensemble activity. Nat Neurosci 21, 881–893 (2018). https://doi.org/10.1038/s41593-018-0139-8
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