| Citation: | LANG Song, ZHANG Yan-wei, ZHENG Han-qing, XU Lin-yu, WANG Lu-han, GONG Yan. Wide-field-of-view and high-resolution HiLo optical sectioning microscopy system[J].Chinese Optics, 2022, 15(6): 1302-1312.doi:10.37188/CO.2022-0087
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The fields of modern biology and biomedicine urgently need wide-field-of-view (FOV), high-resolution microscopic technology and instruments for trans-scale observation of biological samples to meet the requirement of major scientific for research. Limited by the spatial bandwidth product, traditional commercial microscopes cannot meet this demand. Besides, the existing high spatial bandwidth product microscopy systems have problems such as bulky volume and high implementation costs. In this paper, based on the HiLo optical sectioning technology and the self-designed wide-field-of-view and high-resolution objective, a wide-field-of-view and high-resolution HiLo optical sectioning microscopy system was developed. The FOV and imaging resolution of this system were tested. Brightfield imaging experiments were carried out on mouse brain slices by this system and the results were compared with that of OLYMPUS commercial microscope. At the same time, wide-field fluorescence imaging comparison experiments were carried out on wheat seed fluorescent slices. The experiment results show that the FOV of this system reaches 4.8 mm×3.6 mm (the diagonal FOV is 6.0 mm), the lateral resolution reaches 0.74 μm, and the axial resolution reaches 4.16 μm. The comparative experiment proved that this system has the advantages of wide FOV, high resolution and the ability of fast optical sectioning imaging simultaneously. This system can carry out rapid 3D imaging of large-volume biological samples, which will provide strong technical support for researches such as embryonic development, brain imaging, and digital pathology diagnosis.
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