PROTOCOL FOR 3-D VISUALIZATION OF MOLECULES ON MICA VIA THE QUICK-FREEZE, DEEP-ETCH TECHNIQUE
PROTOCOL FOR 3-D VISUALIZATION OF MOLECULES ON MICA VIA THE QUICK-FREEZE, DEEP-ETCH TECHNIQUE
复制标题
DOI：
10.1002/jemt.1060130310
复制
发表时间：
1989-11-01
期刊：
影响因子：
--
通讯作者：
HEUSER, J
中科院分区：
文献类型：
--
作者：
HEUSER, J
通过对吸附在云母基质上的分子进行冷冻干燥，分子成像通常比其他方法提供更好的分子图像；与冷冻薄膜或阴性染色相比，图像更容易解释，三维信息含量更高。完整的程序生产和检查的铂-碳分子复制品的描述。主题包括云母片悬浮液的生产，片的化学预处理以增强吸附，样品在云母上的快速冷冻，冷冻断裂设备的最佳操作，以及复制地形的定向。详细分析了立体显微照片的制作，重点是解释所需的摄影程序和正确显微照片方向的识别。提供了从观察到的铂复制品的分子大小到预期分子量的外推准则。
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.