Abstract
We present an Oblique Plane Microscope 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.
Competing Interest Statement
K.M.D. and R.F. have an investment interest in Discovery Imaging Systems, LLC. C.L.A. serves in an advisory role for Novartis, which has an investment interest in alpelisib. A.B.H. receives research grant support from Takeda. C.L.A. receives or has received research grants from Puma Biotechnology, Pfizer, Lilly, Bayer, Takeda, and Radius; holds stock options in Provista 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. There are no other competing interests.
Footnotes
Author list extended. New resolution measurements added. PSF comparison of OPM, square lattice and spinning disk added. PSF over depth figure added. Optical transfer functions added. New transmission measurements added. Wavefront measurements added. New optogenetics experiments and statistical analysis added. Drosophila embryo imaging added. Neuron imaging added. New appendices added. Imaging of mouse brain and lung tissue added. Updated schematic of illumination train added.