Zernike Phase Contrast Cryo-Electron Microscopy and Tomography for Structure Determination at Nanometer and Subnanometer Resolutions
Zernike Phase Contrast Cryo-Electron Microscopy and Tomography for Structure Determination at Nanometer and Subnanometer Resolutions
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DOI：
10.1016/j.str.2010.06.006
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发表时间：
2010-08-11
期刊：
影响因子：
5.7
通讯作者：
Chiu, Wah
中科院分区：
文献类型：
--
作者：
Murata, Kazuyoshi;Liu, Xiangan;Chiu, Wah
泽尼克相衬冷冻电子显微镜（ZPC-cryoEM）是一种新兴的技术，能够产生比传统冷冻电镜更高的图像对比度。通过将该技术与先进的图像处理方法相结合，我们实现了二维细菌视紫红质晶体和epsilon15噬菌体两种生物标本的亚纳米分辨率。对于epsilon15噬菌体的非对称重建，ZPC-cryoEM与传统的cryoEM相比，可以将所需的数据量减少约3倍。在不需要校正对比度传递函数的情况下，以13埃分辨率进行重建。epsilon15噬菌体门静脉顶点的新结构特征在此重建中被揭示。使用ZPC低温电子断层扫描技术（ZPC-cryoET），可以获得与传统低温电子断层扫描相比具有相似水平的数据缩减和更高分辨率的epsilon15噬菌体结构。这些结果显示了ZPC-cryoEM和ZPC-cryoET在纳米和亚纳米分辨率下对大分子机器结构测定的好处。
Zernike phase contrast cryo-electron microscopy (ZPC-cryoEM) is an emerging technique that is capable of producing higher image contrast than conventional cryoEM. By combining this technique with advanced image processing methods, we achieved subnanometer resolution for two biological specimens: 2D bacteriorhodopsin crystal and epsilon15 bacteriophage. For an asymmetric reconstruction of epsilon15 bacteriophage, ZPC-cryoEM can reduce the required amount of data by a factor of similar to 3, compared with conventional cryoEM. The reconstruction was carried out to 13 angstrom resolution without the need to correct the contrast transfer function. New structural features at the portal vertex of the epsilon15 bacteriophage are revealed in this reconstruction. Using ZPC cryo-electron tomography (ZPC-cryoET), a similar level of data reduction and higher resolution structures of epsilon15 bacteriophage can be obtained relative to conventional cryoET. These results show quantitatively the benefits of ZPC-cryoEM and ZPC-cryoET for structural determinations of macromolecular machines at nanometer and subnanometer resolutions.