Schwarz Heinz, Humbel Bruno M
Max Planck Institute for Developmental Biology, Tübingen, Germany.
Methods Mol Biol. 2014;1117:559-92. doi: 10.1007/978-1-62703-776-1_25.
In correlative microscopy, light microscopy provides the overview and orientation of the complex cells and tissue, while electron microscopy offers the detailed localization and correlation of subcellular structures. In this chapter we offer detailed high-quality electron microscopical preparation methods for optimum preservation of the cellular ultrastructure. From such preparations serial thin sections are collected and used for comparative histochemical, immunofluorescence, and immunogold staining.In light microscopy histological stains identify the orientation of the sample and immunofluorescence labeling facilitates to find the region of interest, namely, the labeled cells expressing the macromolecule under investigation. Sections, labeled with immunogold are analyzed by electron microscopy in order to identify the label within the cellular architecture at high resolution.
在相关显微镜检查中,光学显微镜提供复杂细胞和组织的整体概况及定位,而电子显微镜则能实现亚细胞结构的详细定位及关联。在本章中,我们提供了详细的高质量电子显微镜制备方法,以实现细胞超微结构的最佳保存。从这些制备物中收集连续的薄切片,并用于比较组织化学、免疫荧光和免疫金染色。在光学显微镜下,组织学染色可确定样品的方向,免疫荧光标记有助于找到感兴趣的区域,即表达所研究大分子的标记细胞。用免疫金标记的切片通过电子显微镜进行分析,以便在细胞结构内高分辨率地识别标记。