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Effect of fringe-artifact correction on sub-tomogram averaging from Zernike phase-plate cryo-TEM

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Danev,  Radostin
Baumeister, Wolfgang / Molecular Structural Biology, Max Planck Institute of Biochemistry, Max Planck Society;

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Citation

Kishchenko, G. P., Danev, R., Fisher, R., He, J., Hsieh, C., Marko, M., et al. (2015). Effect of fringe-artifact correction on sub-tomogram averaging from Zernike phase-plate cryo-TEM. JOURNAL OF STRUCTURAL BIOLOGY, 191(3), 299-305. doi:10.1016/j.jsb.2015.07.009.


Cite as: https://hdl.handle.net/11858/00-001M-0000-0028-79B3-F
Abstract
Zernike phase-plate (ZPP) imaging greatly increases contrast in cryo-electron microscopy, however fringe artifacts appear in the images. A computational de-fringing method has been proposed, but it has not been widely employed, perhaps because the importance of de-fringing has not been clearly demonstrated. For testing purposes, we employed Zernike phase-plate imaging in a cryo-electron tomographic study of radial-spoke complexes attached to microtubule doublets. We found that the contrast enhancement by ZPP imaging made nonlinear denoising insensitive to the filtering parameters, such that simple low-frequency band-pass filtering made the same improvement in map quality. We employed sub-tomogram averaging, which compensates for the effect of the "missing wedge" and considerably improves map quality. We found that fringes (caused by the abrupt cut-on of the central hole in the phase plate) can lead to incorrect representation of a structure that is well-known from the literature. The expected structure was restored by amplitude scaling, as proposed in the literature. Our results show that de-fringing is an important part of image-processing for cryo-electron tomography of macromolecular complexes with ZPP imaging. (C) 2015 Elsevier Inc. All rights reserved.