Hyatt A D, Eaton B T
Commonwealth Scientific and Industrial Research Organisation, Australian Animal Health Laboratory, Victoria.
Electron Microsc Rev. 1990;3(1):1-27. doi: 10.1016/0892-0354(90)90011-g.
Whole mounts of intact virus-infected cells have been used for several decades to examine virus-cell relationships and virus structure. The general concept of studying virus structure in association with the host cell has recently been expanded to reveal interactions between viruses and the cytoskeleton. The procedure permits utilization of immuno-gold protocols using both the transmission and scanning electron microscopes. The grid-cell-culture technique is reviewed to explain how it can be exploited to provide valuable information about virus structure and replication in both diagnostic and research laboratories. The use of the technique at the research level is discussed using bluetongue virus as a model. The procedure can provide basic structural information about intact virions and additional data on the intracellular location of viruses and virus-specific structures and about the mode of virus release from infected cells. Application of immunoelectron microscopy reveals information on the protein composition of not only released virus particles but also cell surface and cytoskeletal-associated viruses and virus-specific structures. Collectively, this simple and physically gentle technique has provided information which would otherwise be difficult to obtain.
几十年来,完整的病毒感染细胞整装标本一直被用于研究病毒与细胞的关系以及病毒结构。最近,将病毒结构与宿主细胞联系起来进行研究的总体概念已得到扩展,以揭示病毒与细胞骨架之间的相互作用。该方法允许在透射电子显微镜和扫描电子显微镜下使用免疫金技术。本文对网格细胞培养技术进行了综述,以解释如何利用该技术在诊断和研究实验室中提供有关病毒结构和复制的有价值信息。以蓝舌病毒为模型讨论了该技术在研究层面的应用。该方法可以提供关于完整病毒粒子的基本结构信息,以及关于病毒在细胞内的定位、病毒特异性结构,以及病毒从感染细胞中释放方式的额外数据。免疫电子显微镜的应用不仅揭示了释放出的病毒粒子的蛋白质组成信息,还揭示了细胞表面和细胞骨架相关病毒及病毒特异性结构的蛋白质组成信息。总的来说,这种简单且对细胞损伤较小的技术提供了用其他方法难以获得的信息。