Automatic wavefront engeneering for intravital two-photon imaging.

Two-photon microscopy is invaluable for $in vivo$ imaging, although its performance is hindered by aberrations induced by thick biological samples. Adaptive optics (AO) introduces a compensatory distortion of the laser wavefront to minimize aberrations and improve image quality. To improve the accessibility of indirect AO, we developed an automatic wavefront correction tool based on genetic algorithm that allows simple control of the compensation process. We showed that, without AO, more laser power is required to obtain the same intensity signal in mice brain samples. This suggests that our tool may reduce the use of high laser power which increases photodamage. Then we exploited our AO solution while performing single penetrating vessel occlusions. The significant improvement of the signal intensity collected from the sample allowed us to better evaluate microglia surveillance.