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首页> 外文期刊>Journal of bacteriology >Freeze-substitution of gram-negative eubacteria: general cell morphology and envelope profiles.
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Freeze-substitution of gram-negative eubacteria: general cell morphology and envelope profiles.

机译：革兰氏阴性真细菌的冷冻替代：一般细胞形态和包膜特征。

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Freeze-substitution was performed on strains of Escherichia coli, Pasteurella multocida, Campylobacter fetus, Vibrio cholerae, Pseudomonas aeruginosa, Pseudomonas putida, Aeromonas salmonicida, Proteus mirabilis, Haemophilus pleuropneumoniae, Caulobacter crescentus, and Leptothrix discophora with a substitution medium composed of 2% osmium tetroxide and 2% uranyl acetate in anhydrous acetone. A thick periplasmic gel ranging from 10.6 to 14.3 nm in width was displayed in E. coli K-12, K30, and His 1 (a K-12 derivative containing the K30 capsule genes), P. multocida, C. fetus, P. putida, A. salmonicida, H. pleuropneumoniae, and P. mirabilis. The other bacteria possessed translucent periplasms in which a thinner peptidoglycan layer was seen. Capsular polysaccharide, evident as electron-dense fibers radiating outward perpendicular to the cell surface, was observed on E. coli K30 and His 1 and P. mirabilis cells. A more random arrangement of fibers forming a netlike structure was apparent surrounding cells of H. pleuropneumoniae. For the first time a capsule, distinct from the sheath, was observed on L. discophora. In all instances, capsular polysaccharide was visualized in the absence of stabilizing agents such as homologous antisera or ruthenium red. Other distinct envelope structures were observed external to the outer membrane including the sheath of L. discophora and the S layers of A. salmonicida A450 and C. crescentus CB15A. We believe that the freeze-substitution technique presents a more accurate image of the structural organization of these cells and that it has revealed complex ultrastructural relationships between cell envelope constituents previously difficult to visualize by more conventional means of preparation.
机译：对大肠杆菌，多杀巴斯德氏菌，弯曲杆菌，胎儿，霍乱弧菌，铜绿假单胞菌，恶臭假单胞菌，鲑鱼气单胞菌，奇异变形杆菌，嗜血性肺炎性肺炎杆菌，弯月孢杆菌的腐殖质（含2％的盘状杆菌）的菌株进行冷冻替代。四氧化物和2％乙酸铀酰在无水丙酮中的溶液。在大肠杆菌K-12，K30和His 1（含有K30胶囊基因的K-12衍生物），多杀疟原虫，胎儿梭状芽胞杆菌（P. multocida），C。fetus，P。恶臭假单胞菌，鲑鱼假单胞菌，胸膜肺炎支原体和奇异假单胞菌。其他细菌具有半透明的周质，其中可见较薄的肽聚糖层。在大肠杆菌K30以及His 1和P. mirabilis细胞上观察到荚膜多糖，表现为电子致密纤维垂直于细胞表面向外辐射。胸膜肺炎链球菌周围的细胞中形成网状结构的纤维的排列更加随机。第一次在盘状乳杆菌上观察到不同于鞘的荚膜。在所有情况下，在没有稳定剂（例如同源抗血清或钌红）的情况下，都可以观察到荚膜多糖。在外膜外部观察到其他明显的包膜结构，包括盘状乳杆菌的鞘和鲑鱼曲霉A450和新月形梭菌CB15A的S层。我们相信，冷冻替代技术提供了这些细胞的结构组织的更准确的图像，并且它揭示了以前难以通过更常规的制备方法观察到的细胞包膜成分之间的复杂超微结构关系。

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