Al-Amoudi Ashraf, Frangakis Achilleas S
Deutsches Zentrum für Neurodegenerative Erkrankungen e.V, Bonn, Germany.
Methods Mol Biol. 2013;961:97-117. doi: 10.1007/978-1-62703-227-8_4.
Cryo-electron tomography of vitreous sections is currently the only method for visualizing the eukaryotic ultrastructure at close to native state with molecular resolution. Here, we describe the detailed procedure of how to prepare suitable vitreous sections from mammalian skin for cryo-electron tomography, how to align the projection images of the tilt-series, and finally how to perform sub-tomogram averaging on macromolecular complexes with periodic arrangement such as desmosomes.
玻璃体切片的冷冻电子断层扫描是目前唯一一种以分子分辨率可视化接近天然状态的真核超微结构的方法。在此，我们描述了如何从哺乳动物皮肤制备适合用于冷冻电子断层扫描的玻璃体切片的详细步骤，如何对齐倾斜系列的投影图像，以及最后如何对具有周期性排列的大分子复合物（如桥粒）进行亚断层平均。
