Scattering correcting wavefront shaping for three-photon microscopy

Three-photon (3P) microscopy is getting traction due to its superior performance in deep tissues. Yet, aberrations and light scattering still pose one of the main limitations in the attainable depth ranges for high-resolution imaging. Here, we show scattering correcting wavefront shaping with a simple continuous optimization algorithm, guided by the integrated 3P fluorescence signal. We demonstrate focusing and imaging behind scattering layers and investigate convergence trajectories for different sample geometries and feedback non-linearities. Furthermore, we show imaging through a mouse skull and demonstrate a novel, to the best of our knowledge, fast phase estimation scheme that substantially increases the speed at which the optimal correction can be found. (c) 2022 Optica Publishing Group

Automated summary

This study demonstrates scattering correcting wavefront shaping using a simple continuous optimization algorithm to improve the performance of three-photon microscopy in deep tissue imaging. The research shows the ability to focus and image behind scattering layers and explores convergence trajectories for different sample geometries and feedback non-linearities. Additionally, a novel fast phase estimation scheme is presented for imaging through a mouse skull.