Journal
MEMBRANES
Volume 11, Issue 8, Pages -Publisher
MDPI
DOI: 10.3390/membranes11080634
Keywords
HiLo microscopy; line scanning temporal focusing microscopy; deep tissue imaging; contrast enhancement; axial confinement enhancement
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Funding
- National Natural Science Foundation of China (NSFC) [61831014, 61771287, 32021002]
- Tsinghua University Initiative Scientific Research Program [20193080076]
- Graduate Education Innovation Grants, Tsinghua University [201905J003]
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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.
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.
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