Abstract
In light-sheet fluorescence microscopy (LSFM) a thin light sheet is used to excite the specimen from the side and imaging is performed in the direction perpendicular to the light-sheet. It has the advantages of fast imaging speed, high optical sectioning capability and low photobleaching and phototoxicity to samples. Therefore, it is suitable for high-quality, long-term three-dimensional dynamic observation of large living biological samples. However, the traditional Gaussian light sheet illumination microscopy technology has the problems of small imaging field of view and low spatial resolution. Based on the existing dual-sided illumination LSFM, a large field of view and high resolution LSFM combined with virtual single-pixel imaging deconvolution is presented in this paper, which improves the field of view and resolution of LSFM simultaneously. The relevant microscope is designed and built, and three-dimensional optical sectioning imaging experiments on fluorescent beads and transgenic zebrafish standard samples are carried out. The experimental results prove the three-dimensional high resolution imaging capability of the microscope, which is of great significance in developing the large field of view and high resolution LSFM.
Highlights
a thin light sheet is used to excite the specimen from the side and imaging is performed in the direction
It has the advantages of fast imaging speed
it is suitable for high-quality
Summary
基于希尔伯特变换的结构光照明快速三维彩色显微成像方法 Fast structured illumination three-dimensional color microscopic imaging method based on Hilbert-transform 物理学报. 基于电动可调焦透镜的大范围快速光片显微成像 High-speed and large-scaled light-sheet microscopy with electrically tunable lens 物理学报. 基于Hadamard矩阵优化排序的快速单像素成像 Fast single-pixel imaging based on optimized reordering Hadamard basis 物理学报. 基于数字微镜器件的数字线扫描荧光显微成像技术 Digital line scanning fluorescence microscopy based on digital micromirror device 物理学报. 光片荧光显微术 (light-sheet fluorescence microscopy, LSFM) 采用薄片光束从侧面激发样品, 在垂直于 光片方向上进行成像, 具有成像速度快、光学层析能力强以及光漂白和光毒性低等优点, 适用于对较大活体 生物样品进行高质量、长时间三维动态观测.
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