Griffiths G, Roos N, Schleich S, Locker J K
European Molecular Biology Laboratory, 69117 Heidelberg, Germany.
J Virol. 2001 Nov;75(22):11056-70. doi: 10.1128/JVI.75.22.11056-11070.2001.
In the preceding study (see accompanying paper), we showed by a variety of different techniques that intracellular mature vaccinia virus (vaccinia IMV) is unexpectedly complex in its structural organization and that this complexity also extends to the underlying viral core, which is highly folded. With that analysis as a foundation, we now present different thin-section electron microscopy approaches for analyzing the IMV and the processes by which it is assembled in infected HeLa cells. We focus on conventional epoxy resin thin sections as well as cryosections to describe key intermediates in the assembly process. We took advantage of streptolysin O's ability to selectively permeabilize the plasma membrane of infected cells to improve membrane contrast, and we used antibodies against bone fide integral membrane proteins of the virus to unequivocally identify membrane profiles in thin sections. All of the images presented here can be rationalized with respect to the model put forward for the assembly of the IMV in the accompanying paper.
在之前的研究（见随附论文）中，我们通过多种不同技术表明，细胞内成熟痘苗病毒（痘苗IMV）的结构组织出人意料地复杂，而且这种复杂性还延伸至高度折叠的潜在病毒核心。基于该分析，我们现在展示用于分析IMV及其在感染的HeLa细胞中组装过程的不同超薄切片电子显微镜方法。我们着重于传统环氧树脂超薄切片以及冷冻切片，以描述组装过程中的关键中间体。我们利用链球菌溶血素O选择性通透感染细胞的质膜以改善膜对比度的能力，并使用针对该病毒真正整合膜蛋白的抗体来明确识别超薄切片中的膜轮廓。此处呈现的所有图像都可以根据随附论文中提出的IMV组装模型进行合理阐释。
