Heuser J
Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, Missouri 63110.
J Electron Microsc Tech. 1989 Nov;13(3):244-63. doi: 10.1002/jemt.1060130310.
Molecular imaging by freeze-drying of molecules adsorbed to a mica substrate often provides better images of molecules than those attainable with other methods; the images are easier to interpret than those obtained with frozen thin film or negative staining, and the 3-dimensional information content is greater. The complete procedure for the production and examination of platinum-carbon replicas of molecules is described. Topics include production of a mica flake suspension, chemical pretreatment of the flakes to enhance adsorption, quick-freezing of the samples on mica, optimal operation of the freeze-fracture equipment, and orientation of replica topography. The production of stereo micrographs is analyzed in detail, with emphasis on the photographic procedures necessary for interpretation and on the identification of correct micrograph orientation. Guidelines are provided for the extrapolation from observed molecular size in platinum replicas to expected molecular weight.
通过冷冻干燥吸附在云母基底上的分子进行分子成像，通常能提供比其他方法更好的分子图像；这些图像比通过冷冻薄膜或负染色获得的图像更易于解读，并且三维信息含量更高。本文描述了分子铂 - 碳复制品的制备和检测的完整程序。主题包括云母片悬浮液的制备、对云母片进行化学预处理以增强吸附、在云母上对样品进行快速冷冻、冷冻断裂设备的最佳操作以及复制品形貌的定向。详细分析了立体显微照片的制作，重点在于解读所需的摄影程序以及正确显微照片方向的识别。提供了从铂复制品中观察到的分子大小推断预期分子量的指导原则。
