High-resolution cryo-EM reconstructions in the presence of substantial aberrations

Research output: Contribution to journalArticle

Abstract

Here, an analysis is performed of how uncorrected antisymmetric aberrations, such as coma and trefoil, affect cryo-EM single-particle reconstruction (SPR) results, and an analytical formula quantifying information loss owing to their presence is inferred that explains why Fourier-shell coefficient-based statistics may report significantly overestimated resolution if these aberrations are not fully corrected. The analysis is validated with reference-based aberration refinement for two cryo-EM SPR data sets acquired with a 200kV microscope in the presence of coma exceeding 40μm, and 2.3 and 2.7Å reconstructions for 144 and 173kDa particles, respectively, were obtained. The results provide a description of an efficient approach for assessing information loss in cryo-EM SPR data acquired in the presence of higher order aberrations, and address inconsistent guidelines regarding the level of aberrations that is acceptable in cryo-EM SPR experiments.

Original languageEnglish (US)
Pages (from-to)445-452
Number of pages8
JournalIUCrJ
Volume7
DOIs
StatePublished - May 1 2020

Keywords

  • 3D reconstruction and image processing
  • automation
  • axial aberrations
  • coma
  • cryo-EM
  • imaging
  • resolution
  • singleparticle cryo-EM
  • structure determination.
  • trefoil
  • validation

ASJC Scopus subject areas

  • Chemistry(all)
  • Biochemistry
  • Materials Science(all)
  • Condensed Matter Physics

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