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

Nat Commun. 2021 May 14;12(1):2847. doi: 10.1038/s41467-021-22006-5.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Algorithms
  • Cell Line
  • Computer Simulation
  • DNA / chemistry
  • DNA / ultrastructure
  • Humans
  • Imaging, Three-Dimensional
  • Macromolecular Substances / chemistry*
  • Macromolecular Substances / ultrastructure*
  • Molecular Conformation
  • Nuclear Pore / chemistry
  • Nuclear Pore / ultrastructure
  • Nuclear Pore Complex Proteins / chemistry
  • Nuclear Pore Complex Proteins / ultrastructure
  • Signal-To-Noise Ratio
  • Single Molecule Imaging / methods*
  • Single Molecule Imaging / statistics & numerical data

Substances

  • Macromolecular Substances
  • NUP107 protein, human
  • Nuclear Pore Complex Proteins
  • nuclear pore complex protein 96
  • DNA