Adaptive optics correction for Bessel beam propagating through spatially varying biological aberrations

Due to its long illumination depth and non-diffractive propagation property, the Bessel beam shows efficient imaging potential for two-photon light sheet microscopy in live biological applications. However, with the increase of illumination depth, the wavefront distortion induced by the organism cannot be ignored anymore, which seriously affects the imaging resolution. In this work, a new adaptive optical correction method is introduced to compensate for the space-variant aberration caused by the organism. A single spatial light modulator device is used to generate the Bessel beam and correct the aberration at the same time. The experiment results show that the axial resolution can be improved from 3.1μm to 2.3μm for a 140 μm Bessel beam. High-resolution 3D imaging of zebrafish is achieved with a field of view as large as 140 μm × 140 μm × 40 μm, demonstrating the potential for fast, high-resolution, large-field 3D imaging, which has important prospects for studying dynamic biological processes, such as embryonic development and neuronal activity.