Costello M Joseph
Department of Cell and Developmental Biology, University of North Carolina at Chapel Hill, 27599, USA.
Ultrastruct Pathol. 2006 Sep-Oct;30(5):361-71. doi: 10.1080/01913120600932735.
A brief summary of current cryo-electron microscopy methods for processing and imaging biological tissues is provided. The main emphasis is given to two preparation procedures: frozen-hydrated samples because of the remarkable success of cryo-electron crystallography in obtaining near atomic resolution of integral membrane proteins, and high-pressure freezing because of the wide applicability for vitrification of large samples of normal and diseased tissues for ultrastructural and immunolabeling analysis. Methods for examining certain samples with a TEM cryo-stage are summarized. This includes an introduction to the relatively new area of cryo-electron tomography, which offers the possibility to observe the three-dimensional structure of subcellular components using only their natural variations in composition to generate contrast.
本文简要总结了当前用于生物组织处理和成像的冷冻电子显微镜方法。主要重点介绍了两种制备方法：冷冻水合样品，这是因为冷冻电子晶体学在获得完整膜蛋白的近原子分辨率方面取得了显著成功；高压冷冻，这是因为它广泛适用于正常和患病组织的大样本玻璃化，用于超微结构和免疫标记分析。还总结了使用透射电子显微镜冷冻台检查某些样品的方法。这包括对冷冻电子断层扫描这一相对较新领域的介绍，该技术仅利用亚细胞成分组成的自然变化来产生对比度，从而有可能观察其三维结构。
