McDowall A W, Chang J J, Freeman R, Lepault J, Walter C A, Dubochet J
J Microsc. 1983 Jul;131(Pt 1):1-9. doi: 10.1111/j.1365-2818.1983.tb04225.x.
The preparation and high resolution observation of frozen hydrated thin sections has been studied by transmission electron microscopy (TEM and STEM) on model systems, including pure water, protein solutions, catalase crystals, myelin sheath and various tissues. The state of the ice is determined by electron diffraction. Mass measurement in the electron microscope is used to determine section thickness and control hydration. An adequate depth of vitrified material for sectioning can be obtained from many biological suspensions or untreated tissues. Frozen hydrated sections around 100 nm thick can be produced under optimal conditions from vitreous ice or from vitrified biological samples. Sectioning, transfer and observation in the electron microscope is feasible without alteration of the sample hydration or its initial vitrification. Biological structures can be preserved and observed down to 10 nm. Under favourable working conditions, specimen compression during sectioning and electron beam damage are the factors limiting high resolution observations.
通过透射电子显微镜（TEM和STEM）对包括纯水、蛋白质溶液、过氧化氢酶晶体、髓鞘和各种组织在内的模型系统进行了冷冻水合薄片的制备和高分辨率观察。冰的状态由电子衍射确定。电子显微镜中的质量测量用于确定切片厚度和控制水合作用。从许多生物悬浮液或未处理的组织中可以获得足够深度的用于切片的玻璃化材料。在最佳条件下，可以从玻璃态冰或玻璃化生物样品中制备出约100nm厚的冷冻水合切片。在不改变样品水合作用或其初始玻璃化状态的情况下，在电子显微镜中进行切片、转移和观察是可行的。生物结构可以保存并观察到10nm。在有利的工作条件下，切片过程中的样品压缩和电子束损伤是限制高分辨率观察的因素。
