使用升级版透射电子显微镜对包膜病毒进行冷冻电镜观察:甲型流感、乙型流感病毒和 SARS-CoV-2

L. Kordyukova, A. V. Moiseenko, T.A. Timofeeva, I.T. Fedyakina
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引用次数: 0

摘要

冷冻电镜(cryo-EM)是研究包膜病毒(人类和动物的危险病原体)结构不可或缺的工具。然而,它需要高度专业化的设备和精心的样品制备。在这项工作中,利用配备了低温转移支架的 JEOL JEM-2100 透射电子显微镜的功能,展示了甲型和乙型流感病毒株以及用 beta-丙内酯灭活的 SARS-CoV-2 的初步低温电子显微镜数据。通过图像分析可以(1) 区分 "空 "病毒颗粒和 "满 "病毒颗粒(包含核头壳);(2) 观察病毒包膜的脂质双分子层;(3) 识别流感病毒表面抗原和与内部脂质单分子层相结合的 M1 蛋白层;(4) 区分灭活的 SARS-CoV-2 病毒表面上不同形态的 S-尖峰。所开发的方法为基础研究和应用研究提供了良好的图像质量。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Cryo-electron microscopy of enveloped viruses using upgraded transmission electron microscope: Influenza type A, B viruses and SARS-CoV-2
Cryo-electron microscopy (cryo-EM) is indispensable for the structural studies of enveloped viruses – dangerous pathogens of humans and animals. Yet, it requires highly specialized equipment as well as careful sample preparation. In this work, the capabilities of transmission electron microscope JEOL JEM-2100 equipped with cryo-transfer holder are used, and preliminary cryo-EM data for influenza A and B virus strains and SARS-CoV-2 inactivated with beta-propiolactone are presented. Image analysis allows: (1) to distinguish “empty” viral particles from “full” ones (containing nucleocapsid); (2) to visualize the lipid bilayer of the viral envelope; (3) identify influenza virus surface antigens and the M1 protein layer combined with the inner lipid monolayer; (4) distinguish different morphology of S-spikes on the surface of inactivated SARS-CoV-2 virions. The developed approach provides good image quality for both fundamental and applied research.
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