Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, United States.
Lyda Hill Department of Bioinformatics, University of Texas Southwestern Medical Center, Dallas, United States.
Elife. 2020 Nov 12;9:e57681. doi: 10.7554/eLife.57681.
We present an oblique plane microscope (OPM) that uses a bespoke glass-tipped tertiary objective to improve the resolution, field of view, and usability over previous variants. Owing to its high numerical aperture optics, this microscope achieves lateral and axial resolutions that are comparable to the square illumination mode of lattice light-sheet microscopy, but in a user friendly and versatile format. Given this performance, we demonstrate high-resolution imaging of clathrin-mediated endocytosis, vimentin, the endoplasmic reticulum, membrane dynamics, and Natural Killer-mediated cytotoxicity. Furthermore, we image biological phenomena that would be otherwise challenging or impossible to perform in a traditional light-sheet microscope geometry, including cell migration through confined spaces within a microfluidic device, subcellular photoactivation of Rac1, diffusion of cytoplasmic rheological tracers at a volumetric rate of 14 Hz, and large field of view imaging of neurons, developing embryos, and centimeter-scale tissue sections.
我们提出了一种斜平面显微镜（OPM），它使用定制的玻璃尖端三级物镜，在分辨率、视野和易用性方面优于以前的变体。由于其高数值孔径光学器件，该显微镜实现了与晶格光片显微镜的正方形照明模式相当的横向和轴向分辨率，但采用了用户友好和多功能的格式。鉴于这种性能，我们演示了网格蛋白介导的内吞作用、波形蛋白、内质网、膜动力学和自然杀伤介导的细胞毒性的高分辨率成像。此外，我们还可以对一些在传统光片显微镜几何结构中难以或不可能进行的生物学现象进行成像，包括细胞通过微流控装置内的狭窄空间的迁移、Rac1 的亚细胞光激活、细胞质流变学示踪剂以 14 Hz 的体率扩散，以及神经元、发育中的胚胎和厘米级组织切片的大视野成像。
