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MINFLUX nanometer-scale 3D imaging and microsecond-range tracking on a common fluorescence microscope

Author

Listed:
  • Roman Schmidt

    (Abberior Instruments GmbH)

  • Tobias Weihs

    (Abberior Instruments GmbH)

  • Christian A. Wurm

    (Abberior Instruments GmbH
    Abberior GmbH)

  • Isabelle Jansen

    (Abberior Instruments GmbH)

  • Jasmin Rehman

    (Abberior GmbH)

  • Steffen J. Sahl

    (Max Planck Institute for Biophysical Chemistry)

  • Stefan W. Hell

    (Max Planck Institute for Biophysical Chemistry
    Max Planck Institute for Medical Research)

Abstract

The recently introduced minimal photon fluxes (MINFLUX) concept pushed the resolution of fluorescence microscopy to molecular dimensions. Initial demonstrations relied on custom made, specialized microscopes, raising the question of the method’s general availability. Here, we show that MINFLUX implemented with a standard microscope stand can attain 1–3 nm resolution in three dimensions, rendering fluorescence microscopy with molecule-scale resolution widely applicable. Advances, such as synchronized electro-optical and galvanometric beam steering and a stabilization that locks the sample position to sub-nanometer precision with respect to the stand, ensure nanometer-precise and accurate real-time localization of individually activated fluorophores. In our MINFLUX imaging of cell- and neurobiological samples, ~800 detected photons suffice to attain a localization precision of 2.2 nm, whereas ~2500 photons yield precisions

Suggested Citation

  • Roman Schmidt & Tobias Weihs & Christian A. Wurm & Isabelle Jansen & Jasmin Rehman & Steffen J. Sahl & Stefan W. Hell, 2021. "MINFLUX nanometer-scale 3D imaging and microsecond-range tracking on a common fluorescence microscope," Nature Communications, Nature, vol. 12(1), pages 1-12, December.
    • Handle: RePEc:nat:natcom:v:12:y:2021:i:1:d:10.1038_s41467-021-21652-z
    DOI: 10.1038/s41467-021-21652-z
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    Cited by:

    1. Jan-Erik Bredfeldt & Joanna Oracz & Kamila A. Kiszka & Thea Moosmayer & Michael Weber & Steffen J. Sahl & Stefan W. Hell, 2024. "Bleaching protection and axial sectioning in fluorescence nanoscopy through two-photon activation at 515 nm," Nature Communications, Nature, vol. 15(1), pages 1-13, December.
    2. Elias Amselem & Bo Broadwater & Tora Hävermark & Magnus Johansson & Johan Elf, 2023. "Real-time single-molecule 3D tracking in E. coli based on cross-entropy minimization," Nature Communications, Nature, vol. 14(1), pages 1-11, December.
    3. Andrea Bucci & Giorgio Tortarolo & Marcus Oliver Held & Luca Bega & Eleonora Perego & Francesco Castagnetti & Irene Bozzoni & Eli Slenders & Giuseppe Vicidomini, 2024. "4D Single-particle tracking with asynchronous read-out single-photon avalanche diode array detector," Nature Communications, Nature, vol. 15(1), pages 1-12, December.

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