HiLo Based Line Scanning Temporal Focusing Microscopy for High-Speed, Deep Tissue Imaging

High-speed, optical-sectioning imaging is highly desired in biomedical studies, as most bio-structures and bio-dynamics are in three-dimensions. Compared to point-scanning techniques, line scanning temporal focusing microscopy (LSTFM) is a promising method that can achieve high temporal resolution while maintaining a deep penetration depth. However, the contrast and axial confinement would still be deteriorated in scattering tissue imaging. Here, we propose a HiLo-based LSTFM, utilizing structured illumination to inhibit the fluorescence background and, thus, enhance the image contrast and axial confinement in deep imaging. We demonstrate the superiority of our method by performing volumetric imaging of neurons and dynamical imaging of microglia in mouse brains in vivo.

Automated summary

High-speed, optical-sectioning imaging is crucial in biomedical studies where most bio-structures are in three-dimensions. The HiLo-based LSTFM method proposed in this study enhances image contrast and axial confinement in deep imaging, demonstrating its superiority for imaging neurons and dynamic microglia in mouse brains.