Baigent C L, Müller G
Bernhard-Nocht-Institute, Department of Virology, Hamburg, F.R.G.
J Microsc. 1990 Apr;158(Pt 1):73-80.
A carbon-based immunocytochemistry (CarBIC) method, whereby the epoxy resin is completely removed from a section prior to immunocytochemistry (IC), is described. The resulting absence of embedding medium allows for an optimal access of both primary antibodies and marker systems to structures at various levels throughout the depth of the section. A carbon film evaporated onto the surface of the section before extraction of the resin forms a base to which the section adheres and maintains structural integrity during subsequent processing. The conventional and accepted electron microscopical (EM) appearance for epoxy-resin-embedded material is regained by re-embedding in Epon before contrasting in the normal manner. The technique is demonstrated using human cytomegalovirus (HCMV)-infected human fibroblasts. HCMV+ serum is the source of primary antibodies and 10-nm protein A-gold (pA-G10) together with rabbit anti-human IgG + IgM is the marker for IC.
本文描述了一种基于碳的免疫细胞化学（CarBIC）方法，即在免疫细胞化学（IC）之前从切片中完全去除环氧树脂。由于不存在包埋介质，使得一抗和标记系统能够在切片整个深度的不同层面上最佳地接触到结构。在提取树脂之前蒸发到切片表面的碳膜形成了一个基底，切片附着在该基底上，并在后续处理过程中保持结构完整性。通过以常规方式进行对比之前重新包埋在Epon中，可恢复环氧树脂包埋材料的传统且公认的电子显微镜（EM）外观。使用人巨细胞病毒（HCMV）感染的人成纤维细胞演示了该技术。HCMV +血清是一抗的来源，10纳米蛋白A-金（pA-G10）与兔抗人IgG + IgM一起是IC的标记物。
