Jaffe J S, Glaeser R M
Ultramicroscopy. 1984;13(4):373-7. doi: 10.1016/0304-3991(84)90003-2.
A method is presented for preserving the high resolution structure of biological membranes in a frozen-hydrated environment for electron microscopy. The technique consists of sandwiching a specimen between two carbon films and then waiting while some of the solvent evaporates. When the solvent layer is judged to be of an appropriate thickness, the specimen is then frozen in liquid nitrogen. The specimen can then be inserted into the precooled stage of an electron microscope. Electron diffraction studies of the purple membrane of Halobacterium halobium recorded at -120 degrees C have shown that the structure can be preserved to a resolution of 3.5 A. The main advantage of this method over previous techniques is that the hydrating conditions can be accurately controlled.
本文介绍了一种在冷冻水合环境中保存生物膜高分辨率结构以用于电子显微镜观察的方法。该技术包括将样品夹在两个碳膜之间，然后等待部分溶剂蒸发。当判断溶剂层厚度合适时，将样品在液氮中冷冻。然后可将样品插入电子显微镜的预冷载物台。在-120℃下对嗜盐菌紫膜进行的电子衍射研究表明，其结构可保存至3.5埃的分辨率。该方法相对于先前技术的主要优点是水合条件可得到精确控制。
