Walther Paul, Wang Li, Liessem Sandra, Frascaroli Giada
Central Electron Microscopy Facility, Ulm University, Ulm D-89081, Germany.
Methods Cell Biol. 2010;96:603-18. doi: 10.1016/S0091-679X(10)96025-1.
In this article we present and discuss three electron microscopical approaches to investigate the pathway of human cytomegalovirus (Herpesviridae) infection in cultivated human macrophages and fibroblasts. These methods include surface scanning electron microscopy and high-pressure freezing combined with freeze-substitution and electron tomography. With this set of methods, basically all steps of the viral cycle can be investigated and documented. These methods are especially helpful to investigate membrane structures, since viral particles of a diameter of 150 nm fit well into a semi-thin section of about 500 nm that can be imaged in three dimensions with STEM tomography.
在本文中，我们展示并讨论了三种电子显微镜方法，用于研究人巨细胞病毒（疱疹病毒科）在培养的人巨噬细胞和成纤维细胞中的感染途径。这些方法包括表面扫描电子显微镜以及高压冷冻结合冷冻置换和电子断层扫描。通过这套方法，基本上可以研究和记录病毒循环的所有步骤。这些方法对于研究膜结构特别有帮助，因为直径为150 nm的病毒颗粒非常适合在约500 nm的半薄切片中成像，而该切片可以通过扫描透射电子显微镜断层扫描进行三维成像。
