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Cell Rep. 2016 Dec 20;17(12):3385-3394. doi: 10.1016/j.celrep.2016.12.004.

Long-Term Optical Access to an Estimated One Million Neurons in the Live Mouse Cortex.

Author information

1
James H. Clark Center for Biomedical Engineering and Sciences, Stanford University, Stanford, CA 94305, USA; CNC Program, Stanford University, Stanford, CA 94305, USA.
2
James H. Clark Center for Biomedical Engineering and Sciences, Stanford University, Stanford, CA 94305, USA; CNC Program, Stanford University, Stanford, CA 94305, USA; Howard Hughes Medical Institute, Stanford University, Stanford, CA 94305, USA.
3
James H. Clark Center for Biomedical Engineering and Sciences, Stanford University, Stanford, CA 94305, USA; CNC Program, Stanford University, Stanford, CA 94305, USA; Allen Institute for Brain Science, Seattle, WA 98109, USA.
4
James H. Clark Center for Biomedical Engineering and Sciences, Stanford University, Stanford, CA 94305, USA.
5
Allen Institute for Brain Science, Seattle, WA 98109, USA.
6
Institute of Neuroscience, University of Oregon, Eugene, OR 97403, USA.
7
James H. Clark Center for Biomedical Engineering and Sciences, Stanford University, Stanford, CA 94305, USA; CNC Program, Stanford University, Stanford, CA 94305, USA; Howard Hughes Medical Institute, Stanford University, Stanford, CA 94305, USA. Electronic address: mschnitz@stanford.edu.

Abstract

A major technological goal in neuroscience is to enable the interrogation of individual cells across the live brain. By creating a curved glass replacement to the dorsal cranium and surgical methods for its installation, we developed a chronic mouse preparation providing optical access to an estimated 800,000-1,100,000 individual neurons across the dorsal surface of neocortex. Post-surgical histological studies revealed comparable glial activation as in control mice. In behaving mice expressing a Ca2+ indicator in cortical pyramidal neurons, we performed Ca2+ imaging across neocortex using an epi-fluorescence macroscope and estimated that 25,000-50,000 individual neurons were accessible per mouse across multiple focal planes. Two-photon microscopy revealed dendritic morphologies throughout neocortex, allowed time-lapse imaging of individual cells, and yielded estimates of >1 million accessible neurons per mouse by serial tiling. This approach supports a variety of optical techniques and enables studies of cells across >30 neocortical areas in behaving mice.

KEYWORDS:

brain imaging; calcium imaging; dendrites; dendritic spines; fluorescence imaging; mouse behavior; neocortex; neural ensembles; two-photon microscopy

PMID:
28009304
PMCID:
PMC5459490
DOI:
10.1016/j.celrep.2016.12.004
[Indexed for MEDLINE]
Free PMC Article

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