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Direct supercritical angle localization microscopy for nanometer 3D superresolution

Author

Listed:
  • Anindita Dasgupta

    (European Molecular Biology Laboratory
    Friedrich-Schiller-University
    Leibniz Institute of Photonic Technology)

  • Joran Deschamps

    (European Molecular Biology Laboratory)

  • Ulf Matti

    (European Molecular Biology Laboratory)

  • Uwe Hübner

    (Leibniz Institute of Photonic Technology)

  • Jan Becker

    (Leibniz Institute of Photonic Technology)

  • Sebastian Strauss

    (Ludwig Maximilian University
    Max Planck Institute of Biochemistry)

  • Ralf Jungmann

    (Ludwig Maximilian University
    Max Planck Institute of Biochemistry)

  • Rainer Heintzmann

    (Leibniz Institute of Photonic Technology
    Friedrich-Schiller-University
    Friedrich-Schiller-University)

  • Jonas Ries

    (European Molecular Biology Laboratory)

Abstract

3D single molecule localization microscopy (SMLM) is an emerging superresolution method for structural cell biology, as it allows probing precise positions of proteins in cellular structures. In supercritical angle localization microscopy (SALM), z-positions of single fluorophores are extracted from the intensity of supercritical angle fluorescence, which strongly depends on their distance to the coverslip. Here, we realize the full potential of SALM and improve its z-resolution by more than four-fold compared to the state-of-the-art by directly splitting supercritical and undercritical emission, using an ultra-high NA objective, and applying fitting routines to extract precise intensities of single emitters. We demonstrate nanometer isotropic localization precision on DNA origami structures, and on clathrin coated vesicles and microtubules in cells, illustrating the potential of SALM for cell biology.

Suggested Citation

  • Anindita Dasgupta & Joran Deschamps & Ulf Matti & Uwe Hübner & Jan Becker & Sebastian Strauss & Ralf Jungmann & Rainer Heintzmann & Jonas Ries, 2021. "Direct supercritical angle localization microscopy for nanometer 3D superresolution," Nature Communications, Nature, vol. 12(1), pages 1-9, December.
    • Handle: RePEc:nat:natcom:v:12:y:2021:i:1:d:10.1038_s41467-021-21333-x
    DOI: 10.1038/s41467-021-21333-x
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    Cited by:

    1. Yiming Li & Wei Shi & Sheng Liu & Ivana Cavka & Yu-Le Wu & Ulf Matti & Decheng Wu & Simone Koehler & Jonas Ries, 2022. "Global fitting for high-accuracy multi-channel single-molecule localization," Nature Communications, Nature, vol. 13(1), pages 1-11, December.

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