3D particle averaging and detection of macromolecular symmetry in localization microscopy

Hamidreza Heydarian, Maarten Joosten, Adrian Przybylski, Florian Schueder, Ralf Jungmann, Ben van Werkhoven, Jan Keller-Findeisen, Jonas Ries, Sjoerd Stallinga, Mark Bates and Bernd Rieger ()
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Hamidreza Heydarian: Delft University of Technology
Maarten Joosten: Delft University of Technology
Adrian Przybylski: Max Planck Institute for Biophysical Chemistry
Florian Schueder: Ludwig Maximilian University
Ralf Jungmann: Ludwig Maximilian University
Ben van Werkhoven: Netherlands eScience Center
Jan Keller-Findeisen: Max Planck Institute for Biophysical Chemistry
Jonas Ries: Cell Biology and Biophysics Unit, European Molecular Biology Laboratory (EMBL)
Sjoerd Stallinga: Delft University of Technology
Mark Bates: Max Planck Institute for Biophysical Chemistry
Bernd Rieger: Delft University of Technology

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

Abstract: Abstract Single molecule localization microscopy offers in principle resolution down to the molecular level, but in practice this is limited primarily by incomplete fluorescent labeling of the structure. This missing information can be completed by merging information from many structurally identical particles. In this work, we present an approach for 3D single particle analysis in localization microscopy which hugely increases signal-to-noise ratio and resolution and enables determining the symmetry groups of macromolecular complexes. Our method does not require a structural template, and handles anisotropic localization uncertainties. We demonstrate 3D reconstructions of DNA-origami tetrahedrons, Nup96 and Nup107 subcomplexes of the nuclear pore complex acquired using multiple single molecule localization microscopy techniques, with their structural symmetry deducted from the data.

Date: 2021
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DOI: 10.1038/s41467-021-22006-5

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