实验动物与比较医学 ›› 2017, Vol. 37 ›› Issue (2): 155-159.DOI: 10.3969/j.issn.1674-5817.2017.02.014

• 论著 • 上一篇    下一篇

三种不同固定液对小鼠组织过碘酸雪夫染色效果比较

徐瑶瑶, 李思琪, 田雪梅   

  1. 华南师范大学生命科学学院, 广州510631
  • 收稿日期:2016-12-28 出版日期:2017-04-25 发布日期:2017-04-25
  • 作者简介:徐瑶瑶(1991-),女,硕士研究生,从事基因敲除小鼠免疫方面的研究,E-mail:xyyscnu@163.com
  • 基金资助:
    广东省重点基金资助项目(2014A020212504)

Comparison on Periodic Acid Schiff Staining Effect on Mice Tissues with Three Different Fixatives

XU Yao-yao, LI Si-qi   

  1. School of Life Sciences, South China Normal University, Guangzhou 510631, China
  • Received:2016-12-28 Online:2017-04-25 Published:2017-04-25

摘要: 目的 比较小鼠肝脏、肾脏、小肠三种组织在质量分数4%多聚甲醛液、Carnoy’s液、Bouin’s液三种固定液不同固定时间下的制片质量和过碘酸雪夫染色(PAS)效果,筛选出一种最为适用的固定方法。方法 取成年雄性C57BL/6小鼠肝脏、肾脏、小肠,分别固定于三种固定液,固定时间分别是质量分数4%多聚甲醛液: 24 h、48 h、72 h; Carnoy’s液: 4 h、8 h、12 h; Bouin’s液:12 h、24 h、48 h,充分固定后,常规制作石蜡切片,然后进行PAS染色,对比观察染色效果。结果 三种固定液对不同组织、不同固定时间下PAS染色结果存在差异。肝脏、肾脏、小肠经Carnoy’s液处理12 h后PAS效果最佳,质量分数4%多聚甲醛液固定24 h基本满足观察检测效果,而经Bouin’s液固定组织染色效果相对较差。结论 PAS反应对固定液和固定时间具有选择性, Carnoy’s 液是组织进行PAS染色的理想固定液,但从固定效果、重复性、安全性、成本等多因素综合考虑,在PAS反应中,质量分数4%多聚甲醛液可以替代Carnoy’s液。

关键词: Carnoy's液, Bouin's液, 多聚甲醛, 过碘酸雪夫(PAS)染色

Abstract: Objective Through comparing the slides qualities and periodic acid Schiff (PAS) staining effects with three different fixatives and fixed time on the normal liver, kidney, small intestine from mice, so as to select the optimum fixed methods. Methods The liver, kidney and small intestine tissue from adult male C57BL/6 mice were harvested, then fixed in three fixatives with different times respectively, the 4% paraformaldehyde fluid with 24 h, 48 h, 72 h, the Carnoy’s fluid with 4 h, 8 h, 12 h, and the Bouin’s fluid with 12 h, 24 h, 48 h. After fixed sufficiently, produced paraffin section and then stained routinely with PAS, to observe the staining effect under microscope. Result Different PAS staining results existed among three fixatives with different fixed time on three kind of tissues. The best staining result was observed in Carnoy’s fluid fixed for 12 h, the 4% paraformaldehyde fixed 48 h can basically met the criteria for observation. the staining result was relatively poor fixed in Bouin’s fluid. Conclusion PAS reaction was selective for fixative and fixed time. Carnoy’s fluid is an ideal fixative, but based on the fixed effect, repeatability, safety, and cost, 4% paraformaldehyde fluid can replace Carnoy’s liquid and widely used in PAS reaction.

Key words: Carnoy fluid, Bouin's fluid, Paraformaldehyde fluid, Periodic acid Schiff (PAS) staining

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