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Nature Protocols

Publication date: 2014-11-01
Pages: 2515 - 2538
Publisher: Nature Publishing Group

Author:

Goldey, G
Roumis, D ; Glickfeld, L ; Kerlin, A ; Reid, R ; Bonin, Vincent ; Schafer, D ; Andermann, M

Keywords:

Science & Technology, Life Sciences & Biomedicine, Biochemical Research Methods, Biochemistry & Molecular Biology, MOUSE VISUAL-CORTEX, NEURAL ACTIVITY, PROJECTIONS, RESOLUTION, Animals, Cerebral Cortex, Craniotomy, Diagnostic Imaging, Electroencephalography, Equipment Design, Implants, Experimental, Mice, Inbred C57BL, Mice, Transgenic, Skull, Wakefulness, 03 Chemical Sciences, 06 Biological Sciences, 11 Medical and Health Sciences, Bioinformatics

Abstract:

Cranial window implants in head-fixed rodents are becoming a preparation of choice for stable optical access to large areas of the cortex over extended periods of time. Here we provide a highly detailed and reliable surgical protocol for a cranial window implantation procedure for chronic wide-field and cellular imaging in awake, head-fixed mice, which enables subsequent window removal and replacement in the weeks and months after the initial craniotomy. This protocol has facilitated awake, chronic imaging in adolescent and adult mice over several months from a large number of cortical brain regions; targeted virus and tracer injections from data obtained using prior awake functional mapping; and functionally targeted two-photon imaging across all cortical layers in awake mice using a microprism attachment to the cranial window. Collectively, these procedures extend the reach of chronic imaging of cortical function and dysfunction in behaving animals.