The small correction volume for conventional wavefront shaping methods limits their application in biological imaging through scattering media. In this paper, we take advantage of conjugate adaptive optics (CAO) and remote focusing (CAORF) to achieve three-dimensional (3D) scanning through a scattering layer with a single correction. Our results show that the proposed system can provide 10 times wider axial field of view compared with a conventional conjugate AO system when 16,384 segments are used on a spatial light modulator. We demonstrate two-photon imaging with CAORF through mouse skull. The fluorescent microspheres embedded under the scattering layers can be clearly observed after applying the correction.
We developed a three-photon adaptive optics add-on to a commercial two-photon laser scanning microscope. We
demonstrated its capability for structural and functional imaging of neurons labeled with genetically encoded red
fluorescent proteins or calcium indicators deep in the living mouse brain with cellular and subcellular resolution.