High-resolution two-photon transcranial imaging of brain using direct wavefront sensing

Imaging of the brain in its native state at high spatial resolution poses major challenges to visualization techniques. Two-photon microscopy integrated with thinned-skull or optical clearing skull technique provides a minimally invasive tool for vivo imaging cortex mice without activating immune response and inducing injury. However, contrast are severely compromised by heterogeneity skull, limiting depth superficial layer. In this work, an optimized configuration adaptive optics two-photon microscope system improved wavefront sensing algorithm proposed accurate correction aberrations induced window tissue. Using system, we achieved subcellular transcranial layer 5 pyramidal neurons up 700 ?m below pia living mice. addition, investigated microglia–plaque interaction Alzheimer’s disease demonstrated high-precision laser dendrotomy single-spine ablation.