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Multiview super-resolution microscopy

bioRxiv (Cold Spring Harbor Laboratory)(2021)

Laboratory of High Resolution Optical Imaging

Cited 62|Views39
Abstract
We enhance the performance of confocal microscopy over imaging scales spanning tens of nanometers to millimeters in space and milliseconds to hours in time, improving volumetric resolution more than 10-fold while simultaneously reducing phototoxicity. We achieve these gains via an integrated, four-pronged approach: 1) developing compact line-scanners that enable sensitive, rapid, diffraction-limited imaging over large areas; 2) combining line-scanning with multiview imaging, developing reconstruction algorithms that improve resolution isotropy and recover signal otherwise lost to scattering; 3) adapting techniques from structured illumination microscopy, achieving super-resolution imaging in densely labeled, thick samples; 4) synergizing deep learning with these advances, further improving imaging speed, resolution and duration. We demonstrate these capabilities on more than twenty distinct fixed and live samples, including protein distributions in single cells; nuclei and developing neurons in Caenorhabditis elegans embryos, larvae, and adults; myoblasts in Drosophila wing imaginal disks; and mouse renal, esophageal, cardiac, and brain tissues. ### Competing Interest Statement Y.W., X.H., P.J.L., and H.S. have filed invention disclosures covering aspects of this work. M.G. and J.D. are employees of Applied Scientific Instrumentation, which manufactures the line-scanning units used in this work.
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