Hiramatsu Y, Uno F, Yoshida M, Hatano Y, Nii S
Department of Virology, Okayama University Medical School, Japan.
J Electron Microsc (Tokyo). 1999;48(6):937-46. doi: 10.1093/oxfordjournals.jmicro.a023768.
Virions of vaccinia and orf viruses were examined by ultrahigh-resolution scanning electron microscopy using a non-coating method. Intracellular mature particles of vaccinia virus appeared to be covered with a net and ultrastructurally their surface consists of many fine ridges and globules, while the surfaces of orf virus mature particles recovered from infected cells consist of spirally running protrusions. The ridge-like structures of vaccinia virus were presumed to correspond to surface tubules shown by negative staining of this virus, while the spiral protrusions of orf virus were presumed to correspond to spiral threads having a criss-cross appearance by the same staining. Using scanning electron microscopy in which the samples were prepared by the conventional method, we observed: (i) many virions, i.e. one or two hundreds, or occasionally more reaching about one thousand particles, of the IHD strain of vaccinia virus, (ii) many or a moderate number of virions, i.e. about one hundred or fewer particles, of the 58 strain of cowpox virus and (iii) rather few virions, i.e. several tens or fewer particles, of the Iwate strain of orf virus on the free surface of each cell infected with these viruses. It must be noted that the number of virions detected considerably differed in respective cells examined. Virus budding was frequently observed at the cell surface of monolayer cells infected with vaccinia virus but it was never detected with cowpox or orf virus, indicating a difference in the mechanism of virus release between vaccinia and the other two viruses. When whole cells infected with vaccinia virus were examined by a combination of high-voltage and scanning electron microscopies, virions on the cell surface and those inside the cells were clearly differentiated. All virions on the cell surface had an envelope, and some of the envelopes had a slack and/or one or more bulges.
采用非镀膜法,通过超高分辨率扫描电子显微镜对痘苗病毒和羊口疮病毒的病毒粒子进行了检测。痘苗病毒的细胞内成熟粒子似乎被一层网覆盖,从超微结构来看,其表面由许多细嵴和小球组成,而从感染细胞中回收的羊口疮病毒成熟粒子的表面则由螺旋状排列的突起组成。痘苗病毒的嵴状结构被认为与该病毒负染显示的表面小管相对应,而羊口疮病毒的螺旋状突起被认为与相同染色下呈现交叉外观的螺旋线相对应。使用传统方法制备样品的扫描电子显微镜,我们观察到:(i)在感染痘苗病毒IHD株的每个细胞的自由表面上有许多病毒粒子(即一百或两百个,偶尔更多,可达约一千个粒子);(ii)在感染牛痘病毒58株的细胞中有许多或中等数量(即约一百个或更少粒子)的病毒粒子;(iii)在感染岩手株羊口疮病毒的细胞中有相当少(即几十或更少粒子)的病毒粒子。必须指出的是,在所检测的各个细胞中检测到的病毒粒子数量差异很大。在感染痘苗病毒的单层细胞的细胞表面经常观察到病毒出芽,但在牛痘或羊口疮病毒感染的细胞中从未检测到,这表明痘苗病毒与其他两种病毒在病毒释放机制上存在差异。当用高压和扫描电子显微镜联合检查感染痘苗病毒的整个细胞时,细胞表面的病毒粒子和细胞内的病毒粒子能明显区分开来。细胞表面的所有病毒粒子都有包膜,并且一些包膜松弛和/或有一个或多个凸起。