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Nat Protoc. 2010 Feb;5(2):201-8. doi: 10.1038/nprot.2009.222. Epub 2010 Jan 14.

Thinned-skull cranial window technique for long-term imaging of the cortex in live mice.

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  • 1Molecular Neurobiology Program, Department of Physiology and Neuroscience, Skirball Institute, New York University School of Medicine, New York, New York, USA.


Imaging neurons, glia and vasculature in the living brain has become an important experimental tool for understanding how the brain works. Here we describe in detail a protocol for imaging cortical structures at high optical resolution through a thinned-skull cranial window in live mice using two-photon laser scanning microscopy (TPLSM). Surgery can be performed within 30-45 min and images can be acquired immediately thereafter. The procedure can be repeated multiple times allowing longitudinal imaging of the cortex over intervals ranging from days to years. Imaging through a thinned-skull cranial window avoids exposure of the meninges and the cortex, thus providing a minimally invasive approach for studying structural and functional changes of cells under normal and pathological conditions in the living brain.

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