Direct supercritical angle localization microscopy for nanometer 3D superresolution

Anindita Dasgupta, Joran Deschamps, Ulf Matti, Uwe Hübner, Jan Becker, Sebastian Strauss, Ralf Jungmann, Rainer Heintzmann and Jonas Ries ()
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Anindita Dasgupta: European Molecular Biology Laboratory
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
Ralf Jungmann: Ludwig Maximilian University
Rainer Heintzmann: Leibniz Institute of Photonic Technology
Jonas Ries: European Molecular Biology Laboratory

Nature Communications, 2021, vol. 12, issue 1, 1-9

Abstract: 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.

Date: 2021
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DOI: 10.1038/s41467-021-21333-x

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