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

机译：条纹伪影校正对Zernike相位板冷冻TEm的亚层断层图像平均化的影响

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摘要

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.
机译：Zernike相板（ZPP）成像可大大提高冷冻电子显微镜的对比度，但是图像中会出现条纹伪影。已经提出了一种计算去模糊方法，但是由于没有清楚地证明去模糊的重要性，所以它尚未被广泛采用。出于测试目的，我们在低温电子层析成像研究中将Zernike相板成像技术应用于附着在微管双峰上的放射状辐照复合物。我们发现通过ZPP成像增强对比度使非线性降噪对滤波参数不敏感，因此简单的低频带通滤波对地图质量也有相同的改善。我们使用了子断层图平均，可以补偿“缺失楔形”的影响并显着提高地图质量。我们发现条纹（由相板中中心孔的突然切开引起）可能导致结构的错误表示，这在文献中是众所周知的。如文献所述，通过幅度缩放恢复了预期的结构。我们的研究结果表明，去模糊是大分子配合物ZPP成像的低温电子层析成像图像处理的重要组成部分。 （C）2015 Elsevier Inc.保留所有权利。

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