Automatic estimation and correction of anisotropic magnification distortion in electron microscopes

Publication Type

Journal Article

Year of Publication

2015

Journal

J Struct Biol

Volume

192

Pagination

204-208

Abstract

We demonstrate a significant anisotropic magnification distortion, found on an FEI Titan Krios microscope and affecting magnifications commonly used for data acquisition on a Gatan K2 Summit detector. We describe a program (mag_distortion_estimate) to automatically estimate anisotropic magnification distortion from a set of images of a standard gold shadowed diffraction grating. We also describe a program (mag_distortion_correct) to correct for the estimated distortion in collected images. We demonstrate that the distortion present on the Titan Krios microscope limits the resolution of a set of rotavirus VP6 images to ~7 Å, which increases to ~3 Å following estimation and correction of the distortion. We also use a 70S ribosome sample to demonstrate that in addition to affecting resolution, magnification distortion can also interfere with the classification of heterogeneous data.

Keywords

Electron Cryo-Microscopy; Resolution; Magnification Anisotropy; Image Correction

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