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

摘要

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