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

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

Research output: Contribution to journalArticlepeer-review

Abstract

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.

Original languageEnglish (US)
Article numbere57681
Pages (from-to)1-39
Number of pages39
JournaleLife
Volume9
DOIs
StatePublished - Oct 2020

ASJC Scopus subject areas

  • Neuroscience(all)
  • Immunology and Microbiology(all)
  • Biochemistry, Genetics and Molecular Biology(all)

Fingerprint Dive into the research topics of 'A versatile oblique plane microscope for large-scale and high-resolution imaging of subcellular dynamics'. Together they form a unique fingerprint.

Cite this