Biotechnology Bulletin ›› 2016, Vol. 32 ›› Issue (6): 130-134.doi: 10.13560/j.cnki.biotech.bull.1985.2016.06.018

• Research report • Previous Articles     Next Articles

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 Online:2016-06-27 Published:2016-06-28

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