Wolosewick J J
J Cell Biol. 1980 Aug;86(2):675-61. doi: 10.1083/jcb.86.2.675.
The cytoplasm of cells from a variety of tissues has been viewed in sections (0.25-1 micrometers) devoid of any embedding resin. Glutaraldehyde- and osmium tetroxide-fixed tissues were infiltrated and embedded in a water-miscible wax, polyethylene glycol (PEG), and subsequently sectioned on dry glass or diamond knives. The PEG matrix was removed and the sections were placed on Formvarcarbon-polylysine-coated grids, dehydrated, dried by the critical-point method, and observed in either the high- or low-voltage electron microscope. Stereoscopic views of cells devoid of embedding resin present an image of cell utrastructure unobscured by electron-scattering resins similar to the image of whole, unembedded critical-point-dried or freeze-dried cultured cells observed by transmission electron microscopy. All organelles, including the cytoskeletal structures, are identified and appear not to have been damaged during processing, although membrane components appear somewhat less distinct. The absence of an embedding matrix eliminates the need for additional staining to increase contrast, unlike the situation with specimens embedded in standard electron-scattering resins. The PEG technique thus appears to be a valuable adjunct to conventional methods for ultrastructural analysis.
已在不含任何包埋树脂的切片(0.25 - 1微米)中观察了来自多种组织的细胞的细胞质。用戊二醛和四氧化锇固定的组织先经渗透,然后包埋于一种与水混溶的蜡——聚乙二醇(PEG)中,随后用干玻璃刀或钻石刀切片。去除PEG基质,将切片置于经福尔马林 - 碳 - 聚赖氨酸包被的载网上,脱水,用临界点干燥法干燥,然后在高电压或低电压电子显微镜下观察。不含包埋树脂的细胞的立体图像呈现出细胞超微结构的图像,该图像未被电子散射树脂遮挡,类似于通过透射电子显微镜观察到的完整的、未包埋的临界点干燥或冷冻干燥的培养细胞的图像。所有细胞器,包括细胞骨架结构,都能被识别,并且在处理过程中似乎未受到损伤,尽管膜成分看起来不太清晰。与包埋于标准电子散射树脂的标本不同,不存在包埋基质就无需额外染色来增加对比度。因此,PEG技术似乎是传统超微结构分析方法的一种有价值的辅助手段。