Hybrid spatio-spectral coherent adaptive compensation for line-scanning temporal focusing microscopy

Abstract

Line-scanning temporal focusing microscopy (LTFM) is promising for various biomedical studies, due to its higher imaging speed compared to the gold-standard point-scanning two-photon microscopy while maintaining the same axial confinement. However, LTFM is susceptible to deteriorated performances in excitation efficiency, penetration depth, and resolutions, as a result of wavefront distortions in deep-tissue imaging. Here, we report the hybrid spatio-spectral coherent adaptive compensation (HSSCAC), the first reported technique to fully correct the wavefront distortions in LTFM. Compared with the conventional hill-climbing method, HSSCAC can fully compensate the aberrations. We demonstrate the performance of HSSCAC in deep imaging of neurons in cleared mouse brains and in vivo dynamic imaging of microglias in living mouse brains.