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Fast two-photon volumetric imaging of an improved voltage indicator reveals electrical activity in deeply located neurons in the awake brain

bioRxiv, ISSN: 2692-8205
2018
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  • Citations
    13
    • Citation Indexes
      13
      • CrossRef
        13

Article Description

Imaging of transmembrane voltage deep in brain tissue with cellular resolution has the potential to reveal information processing by neuronal circuits in living animals with minimal perturbation. Multi-photon voltage imaging in vivo, however, is currently limited by speed and sensitivity of both indicators and imaging methods. Here, we report the engineering of an improved genetically encoded voltage indicator, ASAP3, which exhibits up to 51% fluorescence responses in the physiological voltage range, sub-millisecond activation kinetics, and full responsivity under two-photon illumination. We also introduce an ultrafast local volume excitation (ULOVE) two-photon scanning method to sample ASAP3 signals in awake mice at kilohertz rates with increased stability and sensitivity. ASAP3 and ULOVE allowed continuous single-trial tracking of spikes and subthreshold events for minutes in deep locations, with subcellular resolution, and with repeated sampling over multiple days. By imaging voltage in visual cortex neurons, we found evidence for cell type-dependent subthreshold modulation by locomotion. Thus, ASAP3 and ULOVE enable continuous high-speed high-resolution imaging of electrical activity in deeply located genetically defined neurons during awake behavior.

Bibliographic Details

Chavarha, Mariya; Villette, Vincent; Dimov, Ivan K.; Pradhan, Lagnajeet; Evans, Stephen W.; Shi, Dongqing; Yang, Renzhi; Chamberland, Simon; Bradley, Jonathan; Mathieu, Benjamin; St-Pierre, Francois; Schnitzer, Mark J.; Bi, Guoqiang; Toth, Katalin; Ding, Jun; Dieudonné, Stéphane; Lin, Michael Z.

Cold Spring Harbor Laboratory

Biochemistry, Genetics and Molecular Biology; Agricultural and Biological Sciences; Immunology and Microbiology; Neuroscience; Pharmacology, Toxicology and Pharmaceutics

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