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

Bromberg, Raquel; Guo, Yirui; Borek, Dominika; Otwinowski, Zbyszek

doi:10.1107/S2052252520002444

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

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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 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.

Authors:: Bromberg, Raquel; Guo, Yirui; Borek, Dominika; Otwinowski, Zbyszek

Publication Date:: 2020-03-26

Research Org.:: Ligo Analytics, Dallas, TX (United States)

Sponsoring Org.:: USDOE Office of Science (SC); National Institute of Allergy and Infectious Diseases, National Institutes of Health, Department of Health and Human Services; UT Southwestern Medical Center

OSTI Identifier:: 1616493

Alternate Identifier(s):: OSTI ID: 1773805

Grant/Contract Number:: SC0019600; HHSN272201700060C; RP170644

Resource Type:: Published Article

Journal Name:: IUCrJ

Additional Journal Information:: Journal Name: IUCrJ Journal Volume: 7 Journal Issue: 3; Journal ID: ISSN 2052-2525

Publisher:: International Union of Crystallography (IUCr)

Country of Publication:: United Kingdom

Language:: English

Subject:: 59 BASIC BIOLOGICAL SCIENCES; cryo-EM; axial aberrations; coma; trefoil; resolution; validation; 3D reconstruction and image processing; automation; single-particle cryo-EM; imaging; structure determination

Citation Formats


                    Bromberg, Raquel, Guo, Yirui, Borek, Dominika, and Otwinowski, Zbyszek. High-resolution cryo-EM reconstructions in the presence of substantial aberrations.  United Kingdom: N. p., 2020. 
Web.  doi:10.1107/S2052252520002444.

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                    Bromberg, Raquel, Guo, Yirui, Borek, Dominika, & Otwinowski, Zbyszek. High-resolution cryo-EM reconstructions in the presence of substantial aberrations.  United Kingdom.  https://doi.org/10.1107/S2052252520002444

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                    Bromberg, Raquel, Guo, Yirui, Borek, Dominika, and Otwinowski, Zbyszek. Thu .  
"High-resolution cryo-EM reconstructions in the presence of substantial aberrations".  United Kingdom.  https://doi.org/10.1107/S2052252520002444.

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@article{osti_1616493,

  title        = {High-resolution cryo-EM reconstructions in the presence of substantial aberrations},

  author       = {Bromberg, Raquel and Guo, Yirui and Borek, Dominika and Otwinowski, Zbyszek},

  abstractNote = {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.},

  doi          = {10.1107/S2052252520002444},

  journal      = {IUCrJ},

  number       = 3,

  volume       = 7,

  place        = {United Kingdom},

  year         = {2020},

  month        = {3}

}

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