We propose HiLo based line-scanning temporal focusing microscopy to enhance contrast and axial confinement in deep imaging, and demonstrate its superiority by volumetric imaging of microglia and neurons in mouse brains in vivo.
To stimulate neural ensemble selectively at high resolution, we propose non-convex optimization with spherical aberration compensation algorithm for two-photon optogenetics, which achieves high excitation efficiency and precise axial positioning.
Wide-field fluorescence microscopy (WFFM) is widely adopted in biomedical studies. However, axial resolution in most WFFM is poor due to the absence of optical-sectioning capability. To achieve wide-field optical-sectioning, several methods have been proposed, most of which need at least two images to reconstruct one optical sectioning image. So, the frame rate of current wide-field optical sectioning microscopy is no more than half of that of conventional WFFM, which may not meet the speed requirement of fast biodynamic studies. We introduce a novel high-speed, wide-field optical sectioning method based on local contrast weighting function and two-dimensional Hilbert-Huang transform, in which only one structured image is required to reconstruct an optical sectioning image. In this way, the loss of temporal resolution in conventional wide-field optical sectioning microscopy is compensated. We validated this method with the imaging of mouse brain slices.
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