Protocol for constructing an extensive cranial window utilizing a PEO-CYTOP nanosheet for in vivo wide-field imaging of the mouse brain

Large-scale optical measurements have revealed the anatomical and functional connectivity among brain regions underlying brain functions. Here, we describe how to construct a cranial window utilizing a polyethylene-oxide-coated CYTOP (PEO-CYTOP) nanosheet that suppresses bleeding on the brain surface of mice. We demonstrate in vivo two-photon imaging through the PEO-CYTOP nanosheet at the subcellular resolution in the parietal region of the mouse brain. This protocol improves the surgical procedure and expands the optically observable regions, thereby promoting understanding of brain function. For complete details on the use and execution of this protocol, please refer to Takahashi et al. (2020).