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

Roman Schmidt (), Tobias Weihs, Christian A. Wurm, Isabelle Jansen, Jasmin Rehman, Steffen J. Sahl and Stefan W. Hell ()
Additional contact information
Roman Schmidt: Abberior Instruments GmbH
Tobias Weihs: Abberior Instruments GmbH
Christian A. Wurm: Abberior Instruments 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

Nature Communications, 2021, vol. 12, issue 1, 1-12

Abstract: 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

Date: 2021
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DOI: 10.1038/s41467-021-21652-z

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