三种不同固定液对豚鼠眼球的固定效果比较

摘要

目的:用过碘酸雪夫氏(Periodic Acid-Schiff,PAS)染色法比较4％多聚甲醛固定液、4％戊二醛固定液和Davidson固定液固定豚鼠眼球的固定效果,筛选最佳眼球固定液和固定时间.方法:取正常豚鼠眼球分6组,每组5只,Ⅰ组眼球放入4％多聚甲醛固定液固定24h、Ⅱ组眼球放入4％戊二醛固定液固定24h;Ⅲ～Ⅴ组眼球放入Davidson固定液分别固定3、6、24h;Ⅵ组眼球在Davidson固定液中固定3h后转移到10％中性甲醛中再固定48h.常规制片、PAS染色、显微镜观察,比较不同固定方法对组织的固定效果.结果:Davidson固定液固定3h的固定效果最为理想,Davidson固定液固定3h后转移到10％中性甲醛再固定48h的固定效果与Davidson固定液固定3h的固定效果接近,这两组固定液固定的眼球切片均结构完整、层次清晰,视网膜各层细胞排列整齐.结论:Davidson固定液对豚鼠眼球的固定效果明显优于其他两种固定液,豚鼠眼球用Davidson固定液固定后,可将其转移到中性甲醛中长期保存,其固定效果不受影响.%· AIM:To compare the effect of fixation and select the optimal fixation solution and condition for PAS staining of guinea pig eyes among three different fixation solution:4％ paraformaldehyde solution,4％ glutaraldehyde solution and Davidson solution.· METHODS:Totally 30 healthy guinea pigs were divided into 6 groups:Group Ⅰ-Ⅱ were fixed in 4％ paraformaldehyde solution and 4％ glutaraldehyde solution for 24h,respectively;Group Ⅲ-Ⅴ were fixed in Davidson solution for 3,6 and 24h,respectively;and Group Ⅵ were fixed in Davidson solution for 3h and then transferred into 10％ neutral formaldehyde solution for 48h.All groups were sectioned by routine section method and undergone PAS staining,and then observed by light microscope.· RESULTS:It was found that the group which was fixed by Davidson solution for 3h,remained the most complete structure for PAS staining (Group Ⅲ).While the effect of fixation for the group which was transferred into 10％ neutral formaldehyde solution for preserving for 48h after fixing in Davidson solution for 3h was also acceptable for PAS staining (Group Ⅵ).The retinal cells remained clear and in order for both groups which was mentioned above.· CONCLUSION:The best fixation condition for PAS staining for eyes of guinea pigs is fixation in Davidson solution for 3h among these fixation conditions,while it is also suitable to transfer the eyes into neutral formaldehyde solution after fixing in Davidson solution for 3h for preserving for long periods,which is not severely reduced the effect of fixation for PAS staining.