McDonald Kent L, Sharp David J, Rickoll Wayne
Cold Spring Harb Protoc. 2012 Jul 1;2012(7):810-2. doi: 10.1101/pdb.prot068437.
Postembedding immunolabeling using resin sections is the recommended method for beginners carrying out electron microscopy (EM) immunolabeling. Postembedding labeling refers to labeling on sections, which is a method of gaining access to the interior of the cell without the harshness of detergent or ionic extraction as is performed with preembed labeling. Investigators already familiar with routine EM-sectioning techniques find EM immunolabeling using resin sections easiest to do, as procedures are similar to those used when performing light microscopy (LM) immunolabeling, but using a different resin. In addition, the overall preservation of structure is best in resin compared to use of cryosections or preembed labeling. The most critical component of immunoEM (iEM) is what primary antibody to use. This protocol descibes antibody labeling procedures for postembedding iEM using thin sections of Drosophila tissues.
对于刚开始进行电子显微镜（EM）免疫标记的初学者来说，使用树脂切片进行包埋后免疫标记是推荐的方法。包埋后标记是指在切片上进行标记，这是一种无需像包埋前标记那样使用去污剂或离子提取的苛刻方法就能进入细胞内部的方法。已经熟悉常规EM切片技术的研究人员发现，使用树脂切片进行EM免疫标记最容易操作，因为其步骤与进行光学显微镜（LM）免疫标记时相似，但使用的是不同的树脂。此外，与使用冷冻切片或包埋前标记相比，树脂对结构的整体保存效果最佳。免疫电子显微镜（iEM）最关键的部分是使用哪种一抗。本方案描述了使用果蝇组织薄片进行包埋后iEM的抗体标记程序。
