Scothern Anthea, Garrod David
Faculty of Life Sciences, University of Manchester, Manchester, UK.
Methods Cell Biol. 2008;88:347-66. doi: 10.1016/S0091-679X(08)00418-4.
Desmosomes are intercellular junctions found in epithelia and some other tissues. Their primary function is strong cell-cell adhesion. They also link the intermediate filament (IF) cytoskeletons between cells and have roles in cell signaling, tissue morphogenesis, and wound repair. Because of their size (0.2-0.5 microm), details of their ultrastructure can only be resolved at the electron microscope (EM) level. Desmosomes have been visualized using a variety of ultrastructural techniques including lanthanum infiltration, freeze-fracture, electron tomography, cryo-electron microscopy and immunogold labeling. This chapter describes protocols for conventional transmission electron microscopy and for immunogold labeling of ultrathin cryosections. We also discuss the statistical analysis of immunogold particle distribution for low resolution molecular mapping.
桥粒是存在于上皮组织和其他一些组织中的细胞间连接。它们的主要功能是实现细胞间的强黏附。它们还连接细胞间的中间丝(IF)细胞骨架,并在细胞信号传导、组织形态发生和伤口修复中发挥作用。由于其尺寸(0.2 - 0.5微米),其超微结构细节只能在电子显微镜(EM)水平上分辨出来。已使用多种超微结构技术观察到桥粒,包括镧渗透、冷冻断裂、电子断层扫描、冷冻电子显微镜和免疫金标记。本章描述了常规透射电子显微镜和超薄冷冻切片免疫金标记的实验方案。我们还讨论了用于低分辨率分子图谱分析的免疫金颗粒分布的统计分析。