Near-Atomic Resolution Structure Determination in Over-Focus with Volta Phase Plate by Cs-Corrected Cryo-EM
Near-Atomic Resolution Structure Determination in Over-Focus with Volta Phase Plate by Cs-Corrected Cryo-EM
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DOI：
10.1016/j.str.2017.08.008
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发表时间：
2017-10-03
期刊：
影响因子：
5.7
通讯作者：
Wang, Hong-Wei
中科院分区：
文献类型：
--
作者：
Fan, Xiao;Zhao, Lingyun;Wang, Hong-Wei
Volta相板（VPP）是最近发展起来的一种透射电子显微镜（TEM）设备，它可以显著提高生物样品在低温电子显微镜下的图像对比度，从而提供了以高分辨率解决相对较小的大分子结构的可能性。在这项工作中，我们进行了理论分析，发现在物镜球面像差（Cs）校正的TEM上使用相位板可以获得一些有趣的光学特性，包括大分子的过焦成像。随后，我们在cs校正的TEM上评估了冷冻水合载铁蛋白与VPP的成像策略，并从低聚焦和过聚焦数据集中获得了近原子分辨率的载铁蛋白结构，说明了VPP与cs校正的TEM结合进行高分辨率冷冻电镜的可行性和新的潜力。
Volta phase plate (VPP) is a recently developed transmission electron microscope (TEM) apparatus that can significantly enhance the image contrast of biological samples in cryoelectron microscopy, and therefore provide the possibility to solve structures of relatively small macromolecules at high-resolution. In this work, we performed theoretical analysis and found that using phase plate on objective lens spherical aberration (Cs)-corrected TEM may gain some interesting optical properties, including the over-focus imaging of macromolecules. We subsequently evaluated the imaging strategy of frozen-hydrated apo-ferritin with VPP on a Cs-corrected TEM and obtained the structure of apo-ferritin at near-atomic resolution from both under-and over-focused dataset, illustrating the feasibility and new potential of combining VPP with Cs-corrected TEM for high-resolution cryo-EM.