Keller G A, Tokuyasu K T, Dutton A H, Singer S J
Proc Natl Acad Sci U S A. 1984 Sep;81(18):5744-7. doi: 10.1073/pnas.81.18.5744.
Ultrathin frozen sections are ideal substrates with which to carry out immunolabeling experiments in electron microscopy. However, the ultrastructural delineation in positively stained frozen sections has not been as detailed as in conventionally osmium-stained and plastic-embedded sections. We now describe a simple technique in which immunolabeled ultrathin frozen sections are subsequently treated with osmium tetroxide, dehydrated, and then embedded in plastic by impregnation with a monomer to the thickness of the section, followed by polymerization of the monomer. By this technique ultrastructural definition as good as that of conventional plastic sections is achieved, while the high density and specificity of immunolabeling characteristic of ultrathin frozen sections are retained.
超薄冰冻切片是在电子显微镜下进行免疫标记实验的理想底物。然而，阳性染色冰冻切片中的超微结构描绘不如传统锇染色和塑料包埋切片那样详细。我们现在描述一种简单的技术，即对免疫标记的超薄冰冻切片随后用四氧化锇处理、脱水，然后通过用单体浸渍至切片厚度将其包埋在塑料中，随后单体聚合。通过这种技术，可获得与传统塑料切片一样好的超微结构清晰度，同时保留了超薄冰冻切片特有的免疫标记的高密度和特异性。
