Fast wavefront shaping for two-photon brain imaging with large field of view correction

Abstract

Introduction paragraphIn-vivo optical imaging with diffraction-limited resolution deep inside scattering biological tissues is obtained by non-linear fluorescence microscopy1. Active compensation of tissue-induced aberrations and light scattering through adaptive wavefront correction2,3 further extends depth penetration by restoring high resolution at large depth. However, at large depths those corrections are only valid over a very limited field of view within the angular memory effect4. To overcome this limitation, we introduce an acousto-optic light modulation technique for fluorescence imaging with simultaneous wavefront correction at pixel scan speed. Biaxial wavefront corrections are first learned by adaptive optimization at multiple locations in the image field. During image acquisition, the learned corrections are then switched on-the-fly according to the position of the excitation focus during the raster scan. The proposed microscope is applied to in-vivo transcranial neuron imaging and demonstrates correction of skull-induced aberrations and scattering across large fields of view at 40 kHz data acquisition speed.