Addressing preferred specimen orientation in single-particle cryo-EM through tilting

Nat Methods. 2017 Aug;14(8):793-796. doi: 10.1038/nmeth.4347. Epub 2017 Jul 3.


We present a strategy for tackling preferred specimen orientation in single-particle cryogenic electron microscopy by employing tilts during data collection. We also describe a tool to quantify the resulting directional resolution using 3D Fourier shell correlation volumes. We applied these methods to determine the structures at near-atomic resolution of the influenza hemagglutinin trimer, which adopts a highly preferred specimen orientation, and of ribosomal biogenesis intermediates, which adopt moderately preferred orientations.

MeSH terms

  • Algorithms
  • Cryoelectron Microscopy / methods*
  • Hemagglutinin Glycoproteins, Influenza Virus / ultrastructure*
  • Image Enhancement / methods*
  • Imaging, Three-Dimensional / methods*
  • Molecular Imaging / methods*
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Specimen Handling / methods*


  • Hemagglutinin Glycoproteins, Influenza Virus