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

Publication date: 20 December 2016
Source:Cell Reports, Volume 17, Issue 12
Author(s): Tony Hyun Kim, Yanping Zhang, Jérôme Lecoq, Juergen C. Jung, Jane Li, Hongkui Zeng, Cristopher M. Niell, Mark J. Schnitzer
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.

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Teaser

Kim et al. present a preparation for long-term imaging in which a curved glass window replaces the mouse dorsal cranium. This method enables large-scale Ca²⁺ imaging of neuronal dynamics across neocortex in behaving mice and yields an estimated >1 million optically accessible neurons by two-photon microscopy.


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