title: Long-Term Optical Access to an Estimated One Million Neurons in the Live Mouse Cortex.
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.
authors.affiliation: James H. Clark Center for Biomedical Engineering and Sciences, Stanford University, Stanford, CA 94305, USA; CNC Program, Stanford University, Stanford, CA 94305, USA.
authors.forename: Tony Hyun
authors.initials: TH
authors.lastname: Kim
authors.affiliation: 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.
authors.forename: Yanping
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: 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.
authors.forename: Jérôme
authors.initials: J
authors.lastname: Lecoq
authors.affiliation: James H. Clark Center for Biomedical Engineering and Sciences, Stanford University, Stanford, CA 94305, USA; CNC Program, Stanford University, Stanford, CA 94305, USA.
authors.forename: Juergen C
authors.initials: JC
authors.lastname: Jung
authors.affiliation: James H. Clark Center for Biomedical Engineering and Sciences, Stanford University, Stanford, CA 94305, USA.
authors.forename: Jane
authors.initials: J
authors.lastname: Li
authors.affiliation: Allen Institute for Brain Science, Seattle, WA 98109, USA.
authors.forename: Hongkui
authors.initials: H
authors.lastname: Zeng
authors.affiliation: Institute of Neuroscience, University of Oregon, Eugene, OR 97403, USA.
authors.forename: Cristopher M
authors.initials: CM
authors.lastname: Niell
authors.affiliation: 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.
authors.forename: Mark J
authors.initials: MJ
authors.lastname: Schnitzer
mesh_terms: D000818:Animals; D002118:Calcium / Q000737:chemistry; D003712:Dendrites / Q000648:ultrastructure*; D049229:Dendritic Spines / Q000648:ultrastructure*; D051379:Mice; D008856:Microscopy, Fluorescence; D019579:Neocortex / Q000648:ultrastructure*; D017966:Pyramidal Cells / Q000648:ultrastructure*; D059010:Single-Cell Analysis; D059008:Time-Lapse Imaging
chemical_list: D002118:Calcium
keywords: brain imaging; calcium imaging; dendrites; dendritic spines; fluorescence imaging; mouse behavior; neocortex; neural ensembles; two-photon microscopy