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

Author

Listed:
  • Anna-Karin Gustavsson

    (Stanford University
    Karolinska Institutet)

  • Petar N. Petrov

    (Stanford University)

  • Maurice Y. Lee

    (Stanford University
    Stanford University)

  • Yoav Shechtman

    (Stanford University
    Technion, Israel Institute of Technology)

  • W. E. Moerner

    (Stanford University
    Stanford University)

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.

Suggested Citation

  • Anna-Karin Gustavsson & Petar N. Petrov & Maurice Y. Lee & Yoav Shechtman & W. E. Moerner, 2018. "3D single-molecule super-resolution microscopy with a tilted light sheet," Nature Communications, Nature, vol. 9(1), pages 1-8, December.
    • Handle: RePEc:nat:natcom:v:9:y:2018:i:1:d:10.1038_s41467-017-02563-4
    DOI: 10.1038/s41467-017-02563-4
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    Cited by:

    1. Rong Chen & Xiao Tang & Yuxuan Zhao & Zeyu Shen & Meng Zhang & Yusheng Shen & Tiantian Li & Casper Ho Yin Chung & Lijuan Zhang & Ji Wang & Binbin Cui & Peng Fei & Yusong Guo & Shengwang Du & Shuhuai Y, 2023. "Single-frame deep-learning super-resolution microscopy for intracellular dynamics imaging," Nature Communications, Nature, vol. 14(1), pages 1-17, December.

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