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Nat Methods. 2018 Dec;15(12):1108-1116. doi: 10.1038/s41592-018-0188-7. Epub 2018 Nov 12.

Fast, in vivo voltage imaging using a red fluorescent indicator.

Author information

1
The John B. Pierce Laboratory, New Haven, CT, USA.
2
Department of Cellular and Molecular Physiology, Yale University, New Haven, CT, USA.
3
Department of Neuroscience, Yale University, New Haven, CT, USA.
4
James H. Clark Center, Stanford University, Stanford, CA, USA.
5
CNC Program, Stanford University, Stanford, CA, USA.
6
Howard Hughes Medical Institute, Stanford University, Stanford, CA, USA.
7
The John B. Pierce Laboratory, New Haven, CT, USA. vincent.pieribone@yale.edu.
8
Department of Cellular and Molecular Physiology, Yale University, New Haven, CT, USA. vincent.pieribone@yale.edu.
9
Department of Neuroscience, Yale University, New Haven, CT, USA. vincent.pieribone@yale.edu.

Abstract

Genetically encoded voltage indicators (GEVIs) are emerging optical tools for acquiring brain-wide cell-type-specific functional data at unparalleled temporal resolution. To broaden the application of GEVIs in high-speed multispectral imaging, we used a high-throughput strategy to develop voltage-activated red neuronal activity monitor (VARNAM), a fusion of the fast Acetabularia opsin and the bright red fluorophore mRuby3. Imageable under the modest illumination intensities required by bright green probes (<50 mW mm-2), VARNAM is readily usable in vivo. VARNAM can be combined with blue-shifted optical tools to enable cell-type-specific all-optical electrophysiology and dual-color spike imaging in acute brain slices and live Drosophila. With enhanced sensitivity to subthreshold voltages, VARNAM resolves postsynaptic potentials in slices and cortical and hippocampal rhythms in freely behaving mice. Together, VARNAM lends a new hue to the optical toolbox, opening the door to high-speed in vivo multispectral functional imaging.

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PMID:
30420685
PMCID:
PMC6516062
DOI:
10.1038/s41592-018-0188-7
[Indexed for MEDLINE]
Free PMC Article

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