De-scattering with excitation patterning in temporally-focused microscopy (DEEP- TFM)

Point-scanning two-photon microscopy is used routinely for in vivo, volumetric biological imaging, especially in deep tissues. Despite the excellent penetration depth, a conventional point-scanning two-photon microscopy is slow due to the need for raster scanning and imaging time scales linearly with increasing volume, hampering studies of fast biological dynamics. An attractive alternative to point-scanning geometries is wide-field two-photon microscopy, typically called temporal focusing microscopy (TFM) since optical sectioning is achieved by focusing a beam temporally while maintaining wide-field illumination. However, TFM suffers from scattering in tissue resulting in limited imaging depth. Please click Additional Files below to see the full abstract