小鼠卵冠丘复合体固定和染色方法比较

Comparison of fixation and staining methods for mouse oocyte-corona-cumulus complex

目的寻找最佳小鼠卵冠丘复合体固定和染色方法。方法从小鼠输卵管壶腹部取卵冠丘复合体,分别采用丙酮、乙醚-95%乙醇、AF液和4%多聚甲醛固定卵冠丘复合体,分别进行HE、Masson和PAS染色,观察比较卵冠丘复合体形态和染色效果。结果丙酮固定后,卵母细胞和放射冠结构清晰;乙醚-95%乙醇固定,Masson和PAS染色透明带结构清晰;AF液固定,Masson和PAS染色卵母细胞和放射冠结构清晰;4%多聚甲醛固定,Masson和PAS染色透明带结构清晰。结论乙醚-95%乙醇和4%多聚甲醛固定对小鼠卵冠丘复合体固定效果较好,Masson和PAS对透明带染色效果最佳。

Objective To find a better fixation and staining method for mouse oocyte-corona-cumulus complex.Methods The oocyte-corona-cumulus complex were collected from the ampulla of the fallopian tube of mice,fixed with acetone,ether-95%ethanol,AF solution and 4%paraformaldehyde,respectively,and treated with HE,Masson and PAS staining to observe and compare oocyte-corona-cumulus complex morphology and staining effects.Results After acetone fixation,the structure of oocytes and corona radiata was clear;fixed with ether-95%ethanol,the structure of the zona pellucida stained with Masson and PAS was clear;fixed with AF solution,the structure of the oocytes and corona radiata stained with Masson and PAS was clear;4%paraformaldehyde fixation,Masson and PAS dyed zona pellucida has a clear structure.Conclusion The fixation with ether-95%ethanol and 4%paraformaldehyde has a better fixation effect on oocyte-corona-cumulus complex,and Masson and PAS have the best effect on zona pellucida staining.