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

Heydarian, Hamidreza; Joosten, Maarten; Przybylski, Adrian; Schueder, Florian; Jungmann, Ralf; Werkhoven, Ben van; Keller-Findeisen, Jan; Ries, Jonas; Stallinga, Sjoerd; Bates, Mark und Rieger, Bernd (2021): 3D particle averaging and detection of macromolecular symmetry in localization microscopy. In: Nature Communications, Bd. 12, Nr. 1, 2847

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. Adaptation of current algorithms to 3D SMLM data is currently problematic. Here the authors report a method that increases the signal-to-noise ratio and resolution of 3D single particle analysis in localization microscopy and enables determination of the symmetry groups of macromolecular complexes.

• Dokumententyp: Zeitschriftenartikel
• Fakultät: Physik
• Themengebiete: 500 Naturwissenschaften und Mathematik > 530 Physik
• ISSN: 2041-1723
• Sprache: Englisch