TY - JOUR
T1 - A versatile oblique plane microscope for large-scale and high-resolution imaging of subcellular dynamics
AU - Sapoznik, Etai
AU - Chang, Bo Jui
AU - Huh, Jaewon
AU - Ju, Robert J.
AU - Azarova, Evgenia V.
AU - Pohlkamp, Theresa
AU - Welf, Erik S.
AU - Broadbent, David
AU - Carisey, Alexandre F.
AU - Stehbens, Samantha J.
AU - Lee, Kyung Min
AU - Marín, Arnaldo
AU - Hanker, Ariella B.
AU - Schmidt, Jens C.
AU - Arteaga, Carlos L.
AU - Yang, Bin
AU - Kobayashi, Yoshihiko
AU - Tata, Purushothama Rao
AU - Kruithoff, Rory
AU - Doubrovinski, Konstantin
AU - Shepherd, Douglas P.
AU - Millett-Sikking, Alfred
AU - York, Andrew G.
AU - Dean, Kevin M.
AU - Fiolka, Reto P.
N1 - Funding Information:
Ariella B Hanker: Receives research grant support from Takeda. Carlos L Arteaga: Serves in an advisory role for Novartis, which has an investment interest in alpelisib; receives or has received research grants from Puma Biotechnology, Pfizer, Lilly, Bayer, Takeda, and Radius; holds stock options in Pro-vista and Y-TRAP; serves or has served in an advisory role to Novartis, Immunomedics, Merck, Lilly, Symphogen, Daiichi Sankyo, Radius, Taiho Oncology, H3Biomedicine, OrigiMed, Puma Biotechnology, and Sanofi; and reports scientific advisory board renumeration from the Komen Foundation. Alfred Millett-Sikking, Andrew G York: Employee of Calico Life Sciences LLC. Kevin M Dean, Reto P Fiolka: Has an investment interest in Discovery Imaging Systems, LLC. The other authors declare that no competing interests exist.
Publisher Copyright:
© Sapoznik et al.
PY - 2020/10
Y1 - 2020/10
N2 - 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.
AB - 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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U2 - 10.7554/eLife.57681
DO - 10.7554/eLife.57681
M3 - Article
C2 - 33179596
AN - SCOPUS:85097112672
SN - 2050-084X
VL - 9
SP - 1
EP - 39
JO - eLife
JF - eLife
M1 - e57681
ER -