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Anisotropic Correction of Beam-induced Motion for Improved Single-particle Electron Cryo-microscopy

By Shawn Q. Zheng, Eugene Palovcak, Jean-Paul Armache, Yifan Cheng, David A. Agard

Posted 04 Jul 2016
bioRxiv DOI: 10.1101/061960 (published DOI: 10.1038/nmeth.4193)

Correction of electron beam-induced sample motion is one of the major factors contributing to the recent resolution breakthroughs in cryo-electron microscopy. Improving the accuracy and efficiency of motion correction can lead to further resolution improvement. Based on observations that the electron beam induces doming of the thin vitreous ice layer, we developed an algorithm to correct anisotropic image motion at the single pixel level across the whole frame, suitable for both single particle and tomographic images. Iterative, patch-based motion detection is combined with spatial and temporal constraints and dose weighting. The multi-GPU accelerated program, MotionCor2, is sufficiently fast to keep up with automated data collection. The result is an exceptionally robust strategy that can work on a wide range of data sets, including those very close to focus or with very short integration times, obviating the need for particle polishing. Application significantly improves Thon ring quality and 3D reconstruction resolution.

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