ABSTRACT
The glass-window method might induce astrocytic and microglial activation, while the thin-skull method is considered more physiological, as the brain is left untouched. One of the major goals of neuroscience is to understand how the brain works. Two-photon in vivo imaging is a suitable method for monitoring neuronal function and circuits over long timescales, as it has high spatial and temporal resolution. point spread function (PSF) enlargements due to optical aberrations become problematic when the sizes of observation targets are similar, as subtle changes in PSF size can greatly affect measurements, while large targets, such as cell bodies, are affected. To overcome issues of speed, several solutions have been proposed such as imaging cells of interest via random access scanning, or performing volumetric imaging with Bessel beam or special devices. The craniotomy setup provides blood and oxygen supply from brain tissue and cerebrospinal fluid, mechanical protection by the skull, an immune-privileged environment, optical access that is suitable for chronic observation.
