冷冻切片免疫组化染色最佳固定条件探讨  

Optimal fixation conditions for immunohistochemical staining of frozen sections

作　　者：王璇[1] 吴楠[1] 章如松[1] 魏雪[1] 杨万锐 饶秋[1] 马恒辉[1] WANG Xuan;WU Nan;ZHANG Ru-song;WEI Xue;YANG Wan-rui;RAO Qiu;MA Heng-hui(Department of Pathology,Chinese PLA Nanjing General Hospital,Nɑnjing 210002,China)

机构地区：[1]东部战区总医院(原南京军区南京总医院)病理科

出　　处：《诊断病理学杂志》2019年第7期416-418,422,共4页Chinese Journal of Diagnostic Pathology

基　　金：原南京军区南京总医院院管课题(2016052)

摘　　要：目的分析不同固定液及固定时间对冷冻切片免疫组化染色的影响,寻找冷冻切片免疫组化染色最佳固定条件。方法收集新鲜送检的乳腺、肺、甲状腺手术切除标本共15例,冷冻切片后分别置于4种不同固定液(冷丙酮、95%乙醇、4%中性甲醛液、AAF液)中固定5 min和2 h,并对经4%中性甲醛液和AAF液固定的各2张切片进行热修复,根据组织含有的特定抗原进行相应抗体的免疫组化染色。结果采用4种不同固定液固定冷冻切片,均可满足快速病理诊断的需要,但染色细节存在差异;从5个方面综合分析,AAF液的染色效果优于其他3种固定液,其次为95%乙醇、4%中性甲醛、冷丙酮;就阳性强度而言,冷丙酮最强,其次为AAF液、4%中性甲醛、95%乙醇;冷冻切片固定5 min和2 h,除固定2 h阳性强度略强于固定5 min外,其他方面均无明显差异;4%中性甲醛液固定的切片经热修复后与未修复相比组织脱片破损严重,组织结构不清晰,但细胞恢复正常大小;AAF液固定的切片经热修复后与未修复相比有轻微脱片,其他方面无明显差异。结论冷冻切片进行免疫组化染色,AAF液的固定效果优于其他3种固定液;采用AAF液对冷冻切片固定5 min且不修复,整体操作时间短、染色效果好,是冷冻切片免疫组化染色非常理想的固定条件。Objective To analyze the effect of different fixatives and fixation time on immunohistochemical staining of frozen sections and to find the best fixation conditions for frozen sections. Methods Fifteen fresh specimens of breast, lung and thyroid gland were collected. The frozen sections were fixed in four different fixatives(cold acetone, 95% ethanol, 10% neutral formalin, and AAF solution) for 5 minutes and 2 hours respectively, and two sections fixed with 10% neutral formalin and AFF solution were heat-repaired. Immunohistochemical staining was done for frozen sections according to the specific antigens contained in the tissues. Results Four different fixatives which were used to fix frozen sections could all meet the needs of rapid pathological diagnosis, but the details of immunohistochemical staining were different. From five aspects, the staining effect of AAF solution was better than that of the other three fixations, followed by 95% ethanol, 10% neutral formalin and cold acetone. In terms of positive intensity, cold acetone was the strongest, followed by AAF solution, 10% neutral formalin and 95% ethanol. There was no significant difference in other aspects between frozen sections fixed for 5 min and 2 h, except that the positive intensity of frozen sections fixed for 2 h was slightly stronger than that of frozen sections fixed for 5 min. Compared with the non-retrieval sections, the 10% formalin-fixed sections with antigen retrieval were severely damaged, and the tissue structure was not clear;but the cells returned to normal size. The sections fixed with AAF solution were slightly detached after antigen retrieval compared with those without retrieval, and there was no significant difference in other aspects. Conclusions Immunohistochemical staining of frozen section showed that the fixation effect of AAF solution is better than that of the other three kinds of fixatives. Using AAF solution to fix the frozen sections for 5 minutes without antigen retrieval, the whole operation time is short and the stain

关 键 词：冷冻切片 免疫组化染色 固定 

分 类 号：R446.8[医药卫生—诊断学]