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Photoactivated voltage imaging in tissue with an archaerhodopsin-derived reporter.


ABSTRACT: Photoactivated genetically encoded voltage indicators (GEVIs) have the potential to enable optically sectioned voltage imaging at the intersection of a photoactivation beam and an imaging beam. We developed a pooled high-throughput screen to identify archaerhodopsin mutants with enhanced photoactivation. After screening ~105 cells, we identified a novel GEVI, NovArch, whose one-photon near-infrared fluorescence is reversibly enhanced by weak one-photon blue or two-photon near-infrared excitation. Because the photoactivation leads to fluorescent signals catalytically rather than stoichiometrically, high fluorescence signals, optical sectioning, and high time resolution are achieved simultaneously at modest blue or two-photon laser power. We demonstrate applications of the combined molecular and optical tools to optical mapping of membrane voltage in distal dendrites in acute mouse brain slices and in spontaneously active neurons in vivo.

SUBMITTER: Chien MP 

PROVIDER: S-EPMC8099184 | biostudies-literature | 2021 May

REPOSITORIES: biostudies-literature

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Photoactivated voltage imaging in tissue with an archaerhodopsin-derived reporter.

Chien Miao-Ping MP   Brinks Daan D   Testa-Silva Guilherme G   Tian He H   Phil Brooks F F   Adam Yoav Y   Bloxham Blox B   Gmeiner Benjamin B   Kheifets Simon S   Cohen Adam E AE  

Science advances 20210505 19


Photoactivated genetically encoded voltage indicators (GEVIs) have the potential to enable optically sectioned voltage imaging at the intersection of a photoactivation beam and an imaging beam. We developed a pooled high-throughput screen to identify archaerhodopsin mutants with enhanced photoactivation. After screening ~10<sup>5</sup> cells, we identified a novel GEVI, NovArch, whose one-photon near-infrared fluorescence is reversibly enhanced by weak one-photon blue or two-photon near-infrared  ...[more]

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