High-resolution cryo-EM reconstructions in the presence of substantial aberrations.
3D reconstruction and image processing
automation
axial aberrations
coma
cryo-EM
imaging
resolution
single-particle cryo-EM
structure determination
trefoil
validation
Journal
IUCrJ
ISSN: 2052-2525
Titre abrégé: IUCrJ
Pays: England
ID NLM: 101623101
Informations de publication
Date de publication:
01 May 2020
01 May 2020
Historique:
received:
24
12
2019
accepted:
20
02
2020
entrez:
21
5
2020
pubmed:
21
5
2020
medline:
21
5
2020
Statut:
epublish
Résumé
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 200 kV microscope in the presence of coma exceeding 40 µm, and 2.3 and 2.7 Å reconstructions for 144 and 173 kDa 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.
Identifiants
pubmed: 32431828
doi: 10.1107/S2052252520002444
pii: fq5012
pmc: PMC7201289
doi:
Types de publication
Journal Article
Langues
eng
Pagination
445-452Subventions
Organisme : NIGMS NIH HHS
ID : R01 GM117080
Pays : United States
Organisme : NIGMS NIH HHS
ID : R01 GM118619
Pays : United States
Organisme : NIGMS NIH HHS
ID : R21 GM126406
Pays : United States
Informations de copyright
© Raquel Bromberg et al. 2020.
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