eLife (Nov 2020)
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,
- 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 P Fiolka
Affiliations
- Etai Sapoznik
- ORCiD
- 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
- Bo-Jui Chang
- ORCiD
- Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, United States
- Jaewon Huh
- ORCiD
- 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
- Robert J Ju
- ORCiD
- Institute for Molecular Bioscience, University of Queensland, Queensland, Australia
- Evgenia V Azarova
- ORCiD
- Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, United States
- Theresa Pohlkamp
- ORCiD
- Department of Molecular Genetics, University of Texas Southwestern Medical Center, Dallas, United States
- Erik S Welf
- 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
- David Broadbent
- ORCiD
- Institute for Quantitative Health Sciences and Engineering, Michigan State University, East Lansing, United States
- Alexandre F Carisey
- ORCiD
- William T. Shearer Center for Human Immunobiology, Baylor College of Medicine and Texas Children’s Hospital, Houston, United States
- Samantha J Stehbens
- ORCiD
- Institute for Molecular Bioscience, University of Queensland, Queensland, Australia
- Kyung-Min Lee
- Harold C. Simmons Comprehensive Cancer Center and the Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, United States
- Arnaldo Marín
- Harold C. Simmons Comprehensive Cancer Center and the Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, United States; Department of Basic and Clinical Oncology, Faculty of Medicine, University of Chile, Santiago, Chile
- Ariella B Hanker
- ORCiD
- Harold C. Simmons Comprehensive Cancer Center and the Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, United States
- Jens C Schmidt
- ORCiD
- Institute for Quantitative Health Sciences and Engineering, Michigan State University, East Lansing, United States; Department of Obstetrics, Gynecology, and Reproductive Biology, Michigan State University, East Lansing, United States
- Carlos L Arteaga
- Harold C. Simmons Comprehensive Cancer Center and the Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, United States
- Bin Yang
- Chan Zuckerberg Biohub, San Francisco, United States
- Yoshihiko Kobayashi
- ORCiD
- Department of Cell Biology, Duke University School of Medicine, Durham, United States
- Purushothama Rao Tata
- ORCiD
- Department of Cell Biology, Duke University School of Medicine, Durham, United States
- Rory Kruithoff
- Center for Biological Physics and Department of Physics, Arizona State University, Tempe, United States
- Konstantin Doubrovinski
- Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, United States; Cecil H. and Ida Green Comprehensive Center for Molecular, Computational and Systems Biology, University of Texas Southwestern Medical Center, Dallas, United States
- Douglas P Shepherd
- ORCiD
- Center for Biological Physics and Department of Physics, Arizona State University, Tempe, United States
- Alfred Millett-Sikking
- Calico Life Sciences LLC, South San Francisco, United States
- Andrew G York
- Calico Life Sciences LLC, South San Francisco, United States
- Kevin M Dean
- ORCiD
- Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, United States
- Reto P Fiolka
- ORCiD
- 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
- DOI
- https://doi.org/10.7554/eLife.57681
- Journal volume & issue
-
Vol. 9
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.
Keywords
