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Nat Commun. 2018 Jan 9;9(1):123. doi: 10.1038/s41467-017-02563-4.

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

Author information

1
Department of Chemistry, Stanford University, Stanford, CA, 94305, USA.
2
Department of Biosciences and Nutrition, Karolinska Institutet, Stockholm, SE-17177, Sweden.
3
Biophysics Program, Stanford University, Stanford, CA, 94305, USA.
4
Biomedical Engineering Department, Technion, Israel Institute of Technology, Haifa, 3200003, Israel.
5
Department of Chemistry, Stanford University, Stanford, CA, 94305, USA. wmoerner@stanford.edu.
6
Biophysics Program, Stanford University, Stanford, CA, 94305, USA. wmoerner@stanford.edu.

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.

PMID:
29317629
PMCID:
PMC5760554
DOI:
10.1038/s41467-017-02563-4
[Indexed for MEDLINE]
Free PMC Article

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