Murata Takashi, Karahara Ichirou, Kozuka Toshiaki, Thomas H Giddings, Staehelin L Andrew, Mineyuki Yoshinobu
Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, Japan.
J Electron Microsc (Tokyo). 2002;51(2):133-6. doi: 10.1093/jmicro/51.2.133.
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.
我们优化了在高压冷冻和冷冻置换的植物细胞皮层细胞质中观察微丝、微管和被膜小窝的条件，在烟草根尖和洋葱子叶中，经40℃的OsO4处理后再用5%醋酸铀处理的冷冻置换样品中，可以清晰地看到单个微丝和被膜小窝的超分子结构。单独用醋酸铀处理导致细胞质细胞器染色不佳，而单独用OsO4处理的标本中微丝难以辨别。40℃的OsO4染色步骤后再在4℃用醋酸铀染色的组合，未来应会对更详细的植物细胞骨架/膜研究有用。
