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用于研究病毒-宿主相互作用的3D电子显微镜(EM)和相关光电子显微镜(CLEM)方法。

3D Electron Microscopy (EM) and Correlative Light Electron Microscopy (CLEM) Methods to Study Virus-Host Interactions.

作者信息

Romero-Brey Inés

机构信息

Department of Infectious Diseases, Molecular Virology, University of Heidelberg, Heidelberg, Germany.

出版信息

Methods Mol Biol. 2018;1836:213-236. doi: 10.1007/978-1-4939-8678-1_11.

Abstract

Viruses use different strategies to interact with their host and perform a successful viral infection that results in the formation of new infectious viral particles and their propagation to new hosts. Understanding how viruses interact with their hosts requires the use of high-resolution techniques for the direct visualization of these interactions. Here electron microscopy (EM) methods are described that allow the 3D ultrastructural analysis of virus-infected cells. These methods can be implemented with light microscopy (LM) to certainly allocate virus-infected cells or cells displaying a specific/interesting phenotype caused by the interaction of viral proteins with the cellular machinery. Some sample preparation procedures where LM is integrated, known as correlative light electron microscopy (CLEM), are also explained in this chapter. All of these methods are applicable to any kind of cultured cells, including influenza virus-infected cells.

摘要

病毒采用不同策略与宿主相互作用,以实现成功的病毒感染,从而形成新的传染性病毒颗粒并传播至新宿主。了解病毒如何与宿主相互作用需要使用高分辨率技术直接观察这些相互作用。本文介绍了电子显微镜(EM)方法,可对病毒感染细胞进行三维超微结构分析。这些方法可与光学显微镜(LM)结合使用,以准确识别病毒感染细胞或因病毒蛋白与细胞机制相互作用而呈现特定/有趣表型的细胞。本章还解释了一些整合了LM的样品制备程序,即相关光电子显微镜(CLEM)。所有这些方法都适用于任何类型的培养细胞,包括流感病毒感染的细胞。

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