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Cell. 2019 Dec 12;179(7):1590-1608.e23. doi: 10.1016/j.cell.2019.11.004.

Ultrafast Two-Photon Imaging of a High-Gain Voltage Indicator in Awake Behaving Mice.

Author information

1
Institut de Biologie de l'École Normale Supérieure (IBENS), École Normale Supérieure, CNRS, INSERM, PSL Research University, Paris 75005, France.
2
Department of Neurobiology, Stanford University, Stanford, CA 94305, USA; Department of Bioengineering, Stanford University, Stanford, CA 94305, USA.
3
Department of Neurobiology, Stanford University, Stanford, CA 94305, USA; CNC Program, Stanford University, Stanford, CA 94305, USA.
4
Department of Neurobiology, Stanford University, Stanford, CA 94305, USA.
5
Department of Psychiatry and Neuroscience, CERVO Brain Research Centre, Université Laval, Quebec City, QC G1J 2G3, Canada.
6
Department of Neurobiology, Stanford University, Stanford, CA 94305, USA; School of Life Sciences, University of Science and Technology of China, Hefei 230026, China.
7
Department of Neurosurgery, Stanford University, Stanford, CA 94305, USA; Biology PhD Program, Stanford University, Stanford, CA 94305, USA.
8
Department of Bioengineering, Stanford University, Stanford, CA 94305, USA.
9
Université de Paris, SPPIN - Saints-Pères Paris Institute for the Neurosciences, CNRS, Paris F-75006, France.
10
Department of Neuroscience, Baylor College of Medicine, Houston, TX 77030, USA.
11
CNC Program, Stanford University, Stanford, CA 94305, USA; Howard Hughes Medical Institute, Stanford University, Stanford, CA 94305, USA.
12
School of Life Sciences, University of Science and Technology of China, Hefei 230026, China; CAS Center for Excellence in Brain Science and Intelligence Technology, Shanghai 20031, China.
13
Department of Neurosurgery, Stanford University, Stanford, CA 94305, USA.
14
Institut de Biologie de l'École Normale Supérieure (IBENS), École Normale Supérieure, CNRS, INSERM, PSL Research University, Paris 75005, France. Electronic address: dieudon@biologie.ens.fr.
15
Department of Neurobiology, Stanford University, Stanford, CA 94305, USA; Department of Bioengineering, Stanford University, Stanford, CA 94305, USA. Electronic address: mzlin@stanford.edu.

Abstract

Optical interrogation of voltage in deep brain locations with cellular resolution would be immensely useful for understanding how neuronal circuits process information. Here, we report ASAP3, a genetically encoded voltage indicator with 51% fluorescence modulation by physiological voltages, submillisecond activation kinetics, and full responsivity under two-photon excitation. We also introduce an ultrafast local volume excitation (ULoVE) method for kilohertz-rate two-photon sampling in vivo with increased stability and sensitivity. Combining a soma-targeted ASAP3 variant and ULoVE, we show single-trial tracking of spikes and subthreshold events for minutes in deep locations, with subcellular resolution and with repeated sampling over days. In the visual cortex, we use soma-targeted ASAP3 to illustrate cell-type-dependent subthreshold modulation by locomotion. Thus, ASAP3 and ULoVE enable high-speed optical recording of electrical activity in genetically defined neurons at deep locations during awake behavior.

KEYWORDS:

ASAP3; GEVI; RAMP; ULoVE; action potential; electrophysiology; electroporation; theta oscillation; two-photon; voltage imaging

PMID:
31835034
PMCID:
PMC6941988
[Available on 2020-12-12]
DOI:
10.1016/j.cell.2019.11.004

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