Miranda-Saksena Monica, Boadle Ross, Cunningham Anthony L
Centre for Virus Research, Westmead Millennium Institute, The University of Sydney, Darcy Road, Westmead, NSW, 2145, Australia,
Methods Mol Biol. 2014;1144:223-34. doi: 10.1007/978-1-4939-0428-0_15.
Transmission electron microscopy (TEM) provides the resolution necessary to identify both viruses and subcellular components of cells infected with many types of viruses, including herpes simplex virus. Recognized as a powerful tool in both diagnostic and research-based virology laboratories, TEM has made possible the identification of new viruses and has contributed to the elucidation of virus life cycle and virus-host cell interaction. Whilst there are many sample preparation techniques for TEM, conventional processing using chemical fixation and resin embedding remains a useful technique, available in virtually all EM laboratories, for studying virus/cell ultrastructure. In this chapter, we describe the preparation of herpes simplex virus-infected primary neurons, grown on plastic cover slips, to allow sectioning of neurons and axons in their growth plane. This technique allows TEM examination of cell bodies, axons, growth cones, and varicosities, providing powerful insights into virus-cell interaction.
透射电子显微镜（TEM）提供了识别病毒以及感染多种病毒（包括单纯疱疹病毒）的细胞亚细胞成分所需的分辨率。TEM在诊断和基于研究的病毒学实验室中被公认为一种强大的工具，它使鉴定新病毒成为可能，并有助于阐明病毒生命周期和病毒与宿主细胞的相互作用。虽然有许多用于TEM的样品制备技术，但使用化学固定和树脂包埋的传统处理方法仍然是一种有用的技术，几乎所有电子显微镜实验室都可使用，用于研究病毒/细胞超微结构。在本章中，我们描述了在塑料盖玻片上生长的单纯疱疹病毒感染的原代神经元的制备方法，以便在其生长平面上对神经元和轴突进行切片。该技术允许对细胞体、轴突、生长锥和膨体进行TEM检查，从而深入了解病毒与细胞的相互作用。
