3D single-molecule super-resolution microscopy with a tilted light sheet

Anna-Karin Gustavsson, Petar N. Petrov, Maurice Y. Lee, Yoav Shechtman and W. E. Moerner ()
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Anna-Karin Gustavsson: Stanford University
Petar N. Petrov: Stanford University
Maurice Y. Lee: Stanford University
Yoav Shechtman: Stanford University
W. E. Moerner: Stanford University

Nature Communications, 2018, vol. 9, issue 1, 1-8

Abstract: Abstract Tilted light sheet microscopy with 3D point spread functions (TILT3D) combines a novel, tilted light sheet illumination strategy with long axial range point spread functions (PSFs) for low-background, 3D super-localization of single molecules as well as 3D super-resolution imaging in thick cells. Because the axial positions of the single emitters are encoded in the shape of each single-molecule image rather than in the position or thickness of the light sheet, the light sheet need not be extremely thin. TILT3D is built upon a standard inverted microscope and has minimal custom parts. The result is simple and flexible 3D super-resolution imaging with tens of nm localization precision throughout thick mammalian cells. We validate TILT3D for 3D super-resolution imaging in mammalian cells by imaging mitochondria and the full nuclear lamina using the double-helix PSF for single-molecule detection and the recently developed tetrapod PSFs for fiducial bead tracking and live axial drift correction.

Date: 2018
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DOI: 10.1038/s41467-017-02563-4

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