Navigate this Article



How to cite Favorites Q&A Share your feedback Cited by


关键词: 细胞, 超微结构, 果实


注: 除特别说明,所用试剂均为购自sigma-Aldrich公司。

  1. 刀片 (普通双面刀片,用于切取果肉组织)
  2. 包埋盒
  3. Formvar
  4. 苹果
  5. 多聚甲醛
  6.  戊二醛
  7. PBS (磷酸缓冲液 pH 7.2)
  8. 锇酸
  9. 乙醇
  10. 丙酮
  11. Spur
  12. 前固定液 (见溶液配方)
  13. 1% (w/v) 锇酸 (蒸馏水配制)


  1. 4 °C 冰箱
  2. 70 °C 恒温箱
  3. LKB-8800型切片机
  4. 镍网
  5. 透射电镜



  1. 前固定:将苹果果肉组织切分成2-3 mm3的小块,放入到前固定液 (见溶液配方) 中,抽气并在4 °C下固定组织块4 h。
  2. 冲洗:用4 °C保存的0.1 M PBS 洗三次,每次30 min。
  3. 后固定:1%锇酸4 °C下固定1.5-2 h。
  4. 冲洗:用0.1 M PBS (pH 7.2) 4 °C洗三次,每次30 min。
  5. 脱水
    30%乙醇 (20min)→50%乙醇 (20min)→70%乙醇 (过夜)→80%乙醇 (20min)→90%乙醇 (60min)→100%乙醇 (60min)→100%乙醇 (60min)→100%乙醇 (20min)→丙酮 (20min)→丙酮 (20min) →丙酮 (20 min) 
  6. 置换
    丙酮:Spur 2:1 1 h
    丙酮:Spur 1:1 1 h
    丙酮:Spur 1:2 1 h
    纯Spur 1 h
  7. 浸透:纯Spur浸透24 h以上。
  8. 包埋:Spur包埋。
  9. 70 °C下聚合8 h以上。
  10. 切片:用LKB-8800型切片机制备超薄切片,切片捞在100目以0.3% Formvar膜覆盖的镍网上
  11. 用透射电镜进行细胞超微结构观察。



  1. 前固定液
    4% (v/v) 多聚甲醛
    2.5% (v/v) 戊二醛
    0.1 M PBS, pH 7.2 配制
  2. 1% (w/v) 锇酸


感谢国家自然科学基金 (30700646) 资助。本篇实验方案改编自先前发表的文章 Zhang et al. (2004)。


  1. Zhang, L. Y., Peng, Y. B., Pelleschi-Travier, S., Fan, Y., Lu, Y. F., Lu, Y. M., Gao, X. P., Shen, Y. Y., Delrot, S. and Zhang, D. P. (2004). Evidence for apoplasmic phloem unloading in developing apple fruit. Plant Physiol 135(1): 574-586.
Please login or register for free to view full text
Copyright: © 2018 The Authors; exclusive licensee Bio-protocol LLC.
引用格式:张凌云. (2018). 苹果果实细胞超薄切片的制备及超微结构透射电镜观察. Bio-101: e1010241. DOI: 10.21769/BioProtoc.1010241.
How to cite: Zhang, L. Y. (2018). (2018). Preparation of Ultrathin Section of Apple Fruit and Observation of Cellular Ultrastructure by TEM. Bio-101: e1010241. DOI: 10.21769/BioProtoc.1010241.

Please login to post your questions/comments. Your questions will be directed to the authors of the protocol. The authors will be requested to answer your questions at their earliest convenience. Once your questions are answered, you will be informed using the email address that you register with bio-protocol.
You are highly recommended to post your data including images for the troubleshooting.

You are highly recommended to post your data (images or even videos) for the troubleshooting. For uploading videos, you may need a Google account because Bio-protocol uses YouTube to host videos.

We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.