冰冻切片后剩余组织制作石蜡切片与常规石蜡切片的比较

The Paraffin Section Technique of the Remaining Tissue after Frozen Section

目的探讨利用不同温度的固定方法,固定冰冻组织对制作常规HE染色,甲苯胺蓝染色,免疫组化染色的影响。方法48只SD大鼠的结肠、胃冰冻切片后剩余组织,随机分成两组:低温预固定处理组和常温固定组。低温预固定处理组组织放入预冷的4%多聚甲醛固定,置于4℃冰箱1h,随后移入室温固定12h,常规脱水,石蜡包埋;常温固定组,常规脱水,石蜡包埋,两组分别进行HE染色、甲苯胺蓝染色、免疫组化染色,显微镜下观察。结果HE染色低温预固定处理组镜下组织细胞结构清晰可见,细胞核、细胞质对比鲜明,颜色鲜艳;常温固定组组织中可见较多冰晶造成的空隙。甲苯胺蓝染色、免疫组化染色效果无明显差别。结论低温预固定处理的冰冻组织制作石蜡切片,组织结构优于常温组。

Objective To compare the effects of low temperature fixation ordinary temperature fixation on freezing tissue. Methods The remaining anolonely tissue （colon and stomach） after frozen section of 48 SD rats were randomly divided into 2 groups： low temperature fixation （LTF） and ordinary temperature fixation （OTF）. LTF was treated by 4 ~C 4% paraformaldehyde for 1 hour, fixated in room temperature for 12 hours, dehydration and paraffin imbedding by routine method. OTF was fixation in room temperature, dehydration and paraffin imbed- ding by routine method. Both groups were stained by hematoxylin-eosin staining, oluidine blue staining and immunohistochemicat staining methods respectively, being observed by microscope. Results It was shown that the cellular stucture was clearer in LTF than that in OTF. The colour of the paraffin section was bright in LTF. The contrast of the nuclear and cytoplasm in LTF was clear. We could see more spacing caused by ice crystal in paraffin section of OTF. Conclusion There is no obviously differences in immunohistochemical slices.The study indi- cates that the effect of low temperature fixation of the freezing tissue of paraffin section is better than that of ordinary temperature fixation.