生物技术通报 ›› 2016, Vol. 32 ›› Issue (6): 130-134.doi: 10.13560/j.cnki.biotech.bull.1985.2016.06.018

• 研究报告 • 上一篇    下一篇

正庚烷代替二甲苯在拟南芥花药石蜡切片中的应用

涂磊, 张丽瑶,   

  1. 武汉大学生命科学学院,武汉 430000
  • 收稿日期:2015-09-25 出版日期:2016-06-27 发布日期:2016-06-28
  • 作者简介:涂磊,女,硕士研究生,研究方向:发育生物学;E-mail:1013773409@qq.com
  • 基金资助:
    国家自然科学基金项目(31070284)

Replacing Xylene with n-heptane for Paraffin Section of Arabidopsis thaliana Anther

TU Lei, ZHANG Li-yao   

  1. College of Life Sciences,Wuhan University,Wuhan 430000
  • Received:2015-09-25 Published:2016-06-27 Online:2016-06-28

摘要: 二甲苯是应用最广的一种透明剂,常用于石蜡切片中的脱蜡和透明,但由于其对人体危害大,广大科研工作者一直在探寻更加安全的二甲苯代替物。以不同时期的拟南芥花药为试材,脱蜡和透明均由正庚烷代替二甲苯完成,其余步骤与常规石蜡切片相同。结果显示,正庚烷透明力强而迅速,既能与乙醇、封藏树胶混合,又能溶解石蜡,可作为合适的透明剂和脱蜡剂;组织经正庚烷处理后物理性质良好,浸蜡充分,切片时软硬适中,容易连续切片,而且厚薄均匀,摊片时能在42℃水中自然展开,平整而无皱褶;经甲苯胺蓝染色,细胞着色好,核质分明,结构清晰,组织形态保持完好,与二甲苯作透明剂的切片效果无明显区别。因此,正庚烷可以作为一种新的、相对安全有效的二甲苯替代物,应用于植物组织切片中。

关键词: 正庚烷, 二甲苯, 石蜡切片, 透明剂

Abstract: Xylene is widely used as clearing agent for dewaxing and clearing in paraffin section. However,due to its severe harm to human health,researchers have been seeking adequate substitutes. In this study,Arabidopsis thaliana anthers in various periods were processed with n-heptane replacing xylene as a novel clearing and deparaffinizing agent in paraffin section,and the rest of steps were the same as the conventional paraffin section. The results showed that n-heptane with strong and rapid transparency was able to be mixed with ethanol and gum and dissolve paraffin,thus could be utilized as clearing and dewaxing agent. The tissues processed with n-heptane was in fine physic-chemical property and saturated with paraffin;due to the proper hardness of the section,continuously slicing was easy and slice thickness was uniform,moreover,the section was expanded naturally in the water of 42℃ and was flat and no folds. In addition,the cells stained with toluidine blue were in fine quality of staining,and had a clear definition of the nucleus and the cytoplasm,as well as the morphology and structure of the cells were maintained well,meaning that there were no significant differences from the section with xylene as clearing agent. In conclusion,n-heptane is a novel,safe and efficient substitute replacing xylene in the histological techniques for plants.

Key words: n-heptane, xylene, paraffin section, clearing agent