He Wanzhong, He Yongning
National Institute of Biological Sciences, Beijing, China.
Methods Mol Biol. 2014;1117:445-83. doi: 10.1007/978-1-62703-776-1_20.
Electron tomography (ET) is an emerging electron microscopy (EM) technique for three-dimensional (3D) visualization of molecular arrangements and ultrastructural architectures in organelles, cells, and tissues at 2-10 nm resolution. The 3D tomogram is reconstructed from a series of 2D EM images taken from a single specimen at different projecting orientations. The specimen for ET must be specially prepared to meet the ET imaging requirements, i.e., ultrastructural preservation, specimen thickness, tolerance of electron dose and vacuum, and image contrast. In this chapter, the strategies of specimen preparation of organelles, cells, and tissues and the corresponding EM imaging requirements for ET will be described in detail. In addition, the general procedures tomographic reconstruction and tomogram interpretation will be described.
电子断层扫描(ET)是一种新兴的电子显微镜(EM)技术,用于在2-10纳米分辨率下对细胞器、细胞和组织中的分子排列和超微结构进行三维(3D)可视化。三维断层图像是从单个标本在不同投影方向拍摄的一系列二维EM图像重建而来的。用于ET的标本必须经过特殊制备,以满足ET成像要求,即超微结构保存、标本厚度、电子剂量和真空耐受性以及图像对比度。在本章中,将详细描述细胞器、细胞和组织的标本制备策略以及ET相应的EM成像要求。此外,还将描述断层重建和断层图像解释的一般程序。