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A versatile oblique plane microscope for large-scale and high-resolution imaging of subcellular dynamics

2020; eLife Sciences Publications Ltd; Volume: 9; Linguagem: Inglês

10.7554/elife.57681

2050-084X

Etai Sapoznik, Bo-Jui Chang, Jaewon Huh, Robert J. Ju, Evgenia V. Azarova, Theresa Pohlkamp, Erik S. Welf, David Broadbent, Alexandre F. Carisey, Samantha J. Stehbens, Kyung Min Lee, Arnaldo Marín, Ariella B. Hanker, Jens C. Schmidt, Carlos L. Arteaga, Bin Yang, Yoshihiko Kobayashi, Purushothama Rao Tata, Rory Kruithoff, Konstantin Doubrovinski, Douglas P. Shepherd, Alfred Millett-Sikking, Andrew G. York, Kevin M. Dean, Reto Fiolka,

Cell Image Analysis Techniques

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.