Abstract
Chronic cranial window surgery is a critical procedure for in vivo imaging in neuroscience. Here, we describe our surgical protocol with several subtle improvements that increase the success rate significantly. The window allows high-quality imaging in head-fixed behaving mice within the first week after the surgical procedure and remains clear for months. We used this procedure to prepare mice for intrinsic signal imaging and two-photon imaging of layer 6 neurons in visual cortex. For complete details on the use and execution of this protocol, please refer to Augustinaite and Kuhn (2020).