Goldberg Martin W
Department of Biological and Biomedical Sciences, University of Durham, Durham DH1 3LE, United Kingdom.
Methods Cell Biol. 2008;88:109-30. doi: 10.1016/S0091-679X(08)00407-X.
Scanning electron microscopy (SEM) is a high resolution surface imaging technique. Many biological process and structures occur at surfaces and if antibodies are available, their components can be located within the surface structure. This is usually done in a similar way to immuno-fluorescence, using an unconjugated primary antibody followed by a tagged secondary antibody against the primary. In this case the tag is usually a colloidal gold particle instead of a fluorophore. Therefore it is quite straightforward to adapt an immuno-fluorescence procedure for SEM, as long as certain precautions are followed, as discussed here. Progressing from immuno-fluorescence, which essentially only indicates the position of a protein within the volume of a cell, to immuno-SEM, puts the labeling into the context of cellular structures. The principles and practices of sample preparation, labeling and imaging are described here.
扫描电子显微镜（SEM）是一种高分辨率的表面成像技术。许多生物过程和结构发生在表面，如果有抗体可用，其成分可以定位在表面结构内。这通常以与免疫荧光类似的方式进行，使用未结合的一抗，然后是针对一抗的标记二抗。在这种情况下，标记物通常是胶体金颗粒而不是荧光团。因此，只要遵循这里讨论的某些预防措施，就可以很容易地将免疫荧光程序改编用于扫描电子显微镜。从本质上仅指示蛋白质在细胞体积内位置的免疫荧光发展到免疫扫描电子显微镜，将标记置于细胞结构的背景中。这里描述了样品制备、标记和成像的原理及实践。
