A versatile oblique plane microscope for large-scale and high-resolution imaging of subcellular dynamics
Author
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Sapoznik, Etai
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Chang, Bo-Jui
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Huh, Jaewon
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Ju, Robert J.
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Azarova, Evgenia
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Pohlkamp, Theresa
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Welf, Erik S.
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Broadbent, David
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Carisey, Alexandre F.
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Stehbens, Samantha J.
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Lee, Kyung-Min
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Marín Valdés, Arnaldo
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Hanker, Ariella B.
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Schmidt, Jens C.
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Arteaga, Carlos L.
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Bin, Yang
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Kobayashi, Yoshihiko
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Tata, Purushothama Rao
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Kruithoff, Rory
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Doubrovinski, Konstantin
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Shepherd, Douglas P.
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Millett Sikking, Alfred
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York, Andrew G.
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Dean, Kevin M.
Author
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Fiolka, Reto P.
Admission date
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2021-06-09T20:31:05Z
Available date
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2021-06-09T20:31:05Z
Publication date
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2020
Cita de ítem
dc.identifier.citation
eLife 2020;9:e57681
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Identifier
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10.7554/eLife.57681
Identifier
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https://repositorio.uchile.cl/handle/2250/180073
Abstract
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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.
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Patrocinador
dc.description.sponsorship
Cancer Prevention and Research Institute of Texas
RR160057
United States Department of Health & Human Services
National Institutes of Health (NIH) - USA
R00 GM120386
R01HL068702
R33CA235254
R35GM133522
K25 CA204526
P30 CA142543
1R01MH120131-01A1
1R34NS121873
5P30CA142543
R01GM110066
Damon Runyon Cancer Research Foundation
DFS-24-17
Chan Zuckerberg Initiative
HCA3-0000000196
Australian Research Council
FT190100516
Rebecca L. Cooper Medical Research Foundation
PG2018168
University of Queensland
RM2018002613
Company of Biologists
JCSTF1903138
The Welch Foundation
I-1950-20180324
Human Frontier Science Program
LT000911/2018C