Popov V L, Viron A, Eb F, Léfebvre J F, Orfila J
Zh Mikrobiol Epidemiol Immunobiol. 1979 Mar(3):75-9.
The method of cryoultramicrotomy was adapted for the study of the ultrastructure of HeLa and McCoy cells in monolayer cultures infected with Chlamydia, obligatory intracellular procaryotic parasites, the causative agents of ornithosis (strain Loth) and paratrachoma (strain LB 1). The cryosections were obtained by the fixation of the monolayer with 2.5% glutaraldehyde, by the gradual infiltration of precipitated cells with sucrose (0.6--1.2--1.8--2.3 M) prior to freezing in liquid nitrogen, and by the treatment of sections with 1% aqueous methyl cellulose solution before drying. This method ensured good preservation of both Chlamydia, in intracytoplasmic inclusions and host cells, as well as regular reproducibility of the results. Ultrathin sections showed a considerable polymorphism in the vegetative forms of Chlamydia, which was probably due to the structure of their cell walls. Chlamydia, were found to form small vesicle-like structures in the cavities of inclusions. The cell walls and granules inside the elementary bodies of the causative agent of ornithosis were stained with the use of phosphotungstic acid--HCl, pH 0.5.
冷冻超薄切片法被用于研究感染衣原体的单层培养的海拉细胞和 McCoy 细胞的超微结构。衣原体是专性细胞内寄生的原核生物寄生虫,是鸟疫(Loth 菌株)和副沙眼(LB 1 菌株)的病原体。冷冻切片的制备过程如下:先用 2.5%戊二醛固定单层细胞,在液氮中冷冻前,用蔗糖(0.6 - 1.2 - 1.8 - 2.3 M)逐步渗透沉淀的细胞,干燥前用 1%甲基纤维素水溶液处理切片。该方法确保了衣原体在胞质内包涵体和宿主细胞中的良好保存,以及结果的可重复性。超薄切片显示衣原体营养体形式具有相当大的多态性,这可能归因于其细胞壁的结构。发现衣原体在包涵体腔内形成小泡状结构。用 pH 0.5 的磷钨酸 - HCl 对鸟疫病原体原体内部的细胞壁和颗粒进行染色。