The osmium maceration method is a powerful technique for observing the three-dimensional ultrastructure of cellular organelles by scanning electron microscopy. In the conventional osmium maceration method, tissues are immersed in a diluted osmium tetroxide solution for several days at 20°C to remove soluble cytosolic proteins from the freeze-cracked surface of cells, and the optimal duration of this process is dependent on the cell type. To improve the efficiency of the osmium maceration procedure, we have examined systematically the relationship between the reaction temperature and time of the osmium maceration procedure. Treatment at temperatures higher than 20°C drastically shortened the time required to remove cytosolic proteins from the freeze-cracked surface of specimens with optimal durations for the osmium maceration of hepatocytes at 30, 40, 50 and 60°C being 30, 15, 5 and 1 h, respectively. Considering the stability and reproducibility of the macerated specimens, we concluded that the most appropriate temperature was 30 to 40°C. This rapid osmium maceration procedure was used successfully to observe the 3D ultrastructure of Purkinje cells in the cerebellum and proximal convoluted tubule cells in the kidney. This simple and reproducible rapid osmium maceration protocol should find wide appeal for the 3D analysis of cellular organelles in various cell types.

scanning浸渍法是通过扫描电子显微镜观察细胞器的三维超微结构的有力技术。在传统的mac浸法中，将组织在20°C的四氧化稀释溶液中浸泡几天，以从冻裂的细胞表面去除可溶性胞质蛋白，该过程的最佳持续时间取决于细胞类型。为了提高the浸渍程序的效率，我们系统地检查了反应温度和time浸渍程序时间之间的关系。在高于20°C的温度下进行处理，大大缩短了从冷冻破裂的标本表面去除胞浆蛋白所需的时间，并为30分钟的浸透肝细胞提供了最佳持续时间，40、50和60°C分别为30、15、5和1小时。考虑到浸渍试样的稳定性和再现性，我们得出结论，最合适的温度是30至40°C。这种快速的mac浸渍程序已成功用于观察小脑Purkinje细胞和肾脏近曲小管细胞的3D超微结构。这种简单且可重现的快速浸方案对于各种细胞类型的细胞器的3D分析应具有广泛的吸引力。这种快速的mac浸渍程序已成功用于观察小脑Purkinje细胞和肾脏近曲小管细胞的3D超微结构。这种简单且可重现的快速浸方案对于各种细胞类型的细胞器的3D分析应具有广泛的吸引力。这种快速的mac浸渍程序已成功用于观察小脑Purkinje细胞和肾脏近曲小管细胞的3D超微结构。这种简单且可重现的快速浸方案对于各种细胞类型的细胞器的3D分析应具有广泛的吸引力。