Tizro Parastou, Choi Cecilia, Khanlou Negar
Division of Neuropathology, Department of Pathology and Laboratory Medicine, David Geffen School of Medicine at UCLA, Los Angeles, CA, USA.
Methods Mol Biol. 2019;1897:417-424. doi: 10.1007/978-1-4939-8935-5_33.
Transmission electron microscopy (TEM) is an ideal device to study the internal structure of cells and different types of biological materials, but adverse conditions inside electron microscopes such as damage induced by electron bombardment and vacuum evaporation of structural water necessitates complex preparation methods to survive this environment. In order to introduce the sample into the evacuated microscope column, it should be stabilized and altered to small enough (about 3 mm in diameter) and thin enough parts to permit the transmission of electrons. Depending on applications different thicknesses are required; for example, in biological research studies usually 300-500 nm thickness is indicated. To stabilize the specimen and preserve the sample structures, different preparation methods are used involving different steps based on the type of study and the specimen, although the ultimate goal of all these preparation technics is to maintain the native structure of the sample. In this chapter, we try to explain the series of steps that involve in preparation. Virtually every step can affect the quality of sample, and therefore it is important to execute each step in detail.
透射电子显微镜（TEM）是研究细胞内部结构和不同类型生物材料的理想设备，但电子显微镜内部的不利条件，如电子轰击造成的损伤和结构水的真空蒸发，需要复杂的制备方法才能在这种环境中存活。为了将样品引入真空显微镜柱，应将其稳定并处理成足够小（直径约3毫米）且足够薄的部分，以允许电子透过。根据应用的不同，需要不同的厚度；例如，在生物学研究中，通常要求厚度为300-500纳米。为了稳定标本并保存样品结构，根据研究类型和标本的不同，会使用不同的制备方法，这些方法涉及不同的步骤，尽管所有这些制备技术的最终目标都是保持样品的天然结构。在本章中，我们将尝试解释制备过程中涉及的一系列步骤。实际上，每个步骤都可能影响样品质量，因此详细执行每个步骤非常重要。
