We demonstrate transcutical structural and functional imaging of neurons labeled with genetically encoded red fluorescent proteins and calcium indicators in the living Drosophila brain with cellular and subcellular resolution.

The small correction volume for conventional wavefront shaping methods limits their application in biological imaging through scattering media. We demonstrate large volume wavefront shaping through a scattering layer with a single correction by conjugate adaptive optics and remote focusing (CAORF). The remote focusing module can maintain the conjugation between the adaptive optical (AO) element and the scattering layer during three-dimensional scanning. This new configuration provides a wider correction volume by better utilization of the memory effect in a fast three-dimensional laser scanning microscope. 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 also demonstrate three-dimensional fluorescence imaging, multi-spot patterning through a scattering layer and two-photon imaging through mouse skull tissue.