Improved method for visualizing coated pits, microfilaments, and microtubules in cryofixed and freeze-substituted plant cells.
Improved method for visualizing coated pits, microfilaments, and microtubules in cryofixed and freeze-substituted plant cells.
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用于可视化冷冻固定和冷冻替代植物细胞中的包被凹坑、微丝和微管的改进方法。
DOI：
10.1093/jmicro/51.2.133
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发表时间：
2002
期刊：
影响因子：
--
通讯作者：
Y. Mineyuki
中科院分区：
文献类型：
--
作者：
T. Murata;I. Karahara;Toshiaki Kozuka;H. G. Thomas;L. Staehelin;Y. Mineyuki
我们已经优化了条件可视化微丝，微管，并在皮层细胞质的高压冷冻和冷冻取代的植物细胞，在烟草根尖和洋葱子叶，个别微丝和超分子结构的涂层坑可以清楚地看到在冷冻取代的样品OsO 4在40 ℃，然后由5%的醋酸铀酰处理。用醋酸双氧铀单独处理导致染色不佳的细胞质细胞器，而微丝是难以辨别单独与OsO 4处理的标本。40摄氏度OsO 4染色步骤，然后在4摄氏度的醋酸双氧铀染色的组合应证明是有用的，在未来更详细的植物细胞骨架/膜的研究。
We have optimized the conditions for visualizing microfilaments, microtubules, and coated pits in the cortical cytoplasm of high-pressure frozen and freeze-substituted plant cells, in both tobacco root tips and onion cotyledons, individual microfilaments and the supramolecular structure of coated pits can be seen clearly in freeze-substituted samples treated with OsO4 at 40 degrees C followed by 5% uranyl acetate. Treatment with uranyl acetate alone resulted in poorly stained cytoplasmic organelles, whereas microfilaments were difficult to discern in specimen treated with OsO4 alone. The combination of a 40 degrees C OsO4 staining step followed by staining with uranyl acetate at 4 degrees C should prove useful for more detailed plant cytoskeletal/membrane studies in the future.