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  • Review Article
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Probing neural codes with two-photon holographic optogenetics


Optogenetics ushered in a revolution in how neuroscientists interrogate brain function. Because of technical limitations, the majority of optogenetic studies have used low spatial resolution activation schemes that limit the types of perturbations that can be made. However, neural activity manipulations at finer spatial scales are likely to be important to more fully understand neural computation. Spatially precise multiphoton holographic optogenetics promises to address this challenge and opens up many new classes of experiments that were not previously possible. More specifically, by offering the ability to recreate extremely specific neural activity patterns in both space and time in functionally defined ensembles of neurons, multiphoton holographic optogenetics could allow neuroscientists to reveal fundamental aspects of the neural codes for sensation, cognition and behavior that have been beyond reach. This Review summarizes recent advances in multiphoton holographic optogenetics that substantially expand its capabilities, highlights outstanding technical challenges and provides an overview of the classes of experiments it can execute to test and validate key theoretical models of brain function. Multiphoton holographic optogenetics could substantially accelerate the pace of neuroscience discovery by helping to close the loop between experimental and theoretical neuroscience, leading to fundamental new insights into nervous system function and disorder.

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Fig. 1: Two-photon holographic optogenetics.
Fig. 2: Improving the effective spatial fidelity of multiphoton holographic optogenetics.
Fig. 3: Approaches to extend multiphoton holographic optogenetics.
Fig. 4: Multiple approaches to using multiphoton optogenetics to reveal neural codes underlying behavior.
Fig. 5: Examples of using multiphoton holographic optogenetics to address neural codes and plasticity rules.

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We thank H. Bounds and I. Oldenburg for critical review of the manuscript. This work was supported by the New York Stem Cell Foundation and NIH grants UF1NS107574, R01MH117824 and U19 NS107613. H.A. is a New York Stem Cell Robertson Investigator.

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Correspondence to Hillel Adesnik.

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Competing interests

H.A. is a co-inventor of 3D-SHOT, which is discussed in this Review (US Patent and Trademark Office, provisional patent application no. 62-429,017).

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Peer review information Nature Neuroscience thanks Adam Packer and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

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Adesnik, H., Abdeladim, L. Probing neural codes with two-photon holographic optogenetics. Nat Neurosci 24, 1356–1366 (2021).

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