High-resolution cryo-EM using beam-image shift at 200 keV.

IUCrJ
Jennifer N CashMichael A Cianfrocco

Abstract

Recent advances in single-particle cryo-electron microscopy (cryo-EM) data collection utilize beam-image shift to improve throughput. Despite implementation on 300 keV cryo-EM instruments, it remains unknown how well beam-image-shift data collection affects data quality on 200 keV instruments and the extent to which aberrations can be computationally corrected. To test this, a cryo-EM data set for aldolase was collected at 200 keV using beam-image shift and analyzed. This analysis shows that the instrument beam tilt and particle motion initially limited the resolution to 4.9 Å. After particle polishing and iterative rounds of aberration correction in RELION, a 2.8 Å resolution structure could be obtained. This analysis demonstrates that software correction of microscope aberrations can provide a significant improvement in resolution at 200 keV.

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Citations

May 6, 2021·Acta Crystallographica. Section D, Structural Biology·Jan RheinbergerCristina Paulino
May 11, 2021·Microscopy·Radostin DanevMasahide Kikkawa
Jul 3, 2021·International Journal of Molecular Sciences·Chun-Hsiung WangWei-Hau Chang

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Datasets Mentioned

BETA
EMPIAR-10519

Software Mentioned

MicAssess
ChimeraX
CTFFIND
MotionCor
real
PyMOL Molecular Graphics System
Appion
crYOLO
Phenix
EMPIAR

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