Fujimoto K, Noda T, Fujimoto T
Department of Anatomy, Faculty of Medicine, Kyoto University, Japan.
Histochem Cell Biol. 1997 Jan;107(1):81-4. doi: 10.1007/s004180050091.
We describe a simple method for the quick-freezing/freeze-fracturing of cells in tissues or culture monolayers. Tissue slices or cultured cells were covered with thin copper foil (10-micron-thick), and frozen by smashing them against a liquid helium-cooled copper block. Freeze-fracturing was accomplished by mechanically separating the copper foil from the frozen specimen. The fracture faces were replicated by platinum and carbon. Replicas were processed for conventional electron microscopic observation or cytochemical labeling. This method allows the ultrastructural and cytochemical examination of large areas of fractured membrane without chemical fixation.
我们描述了一种用于快速冷冻/冷冻断裂组织或培养单层细胞的简单方法。用薄铜箔（10微米厚）覆盖组织切片或培养细胞，然后将它们撞击到液氦冷却的铜块上进行冷冻。通过机械方式将铜箔与冷冻标本分离来完成冷冻断裂。用铂和碳对断裂面进行复制。复制物经过处理用于常规电子显微镜观察或细胞化学标记。这种方法无需化学固定即可对大面积断裂膜进行超微结构和细胞化学检查。
