摘要
目的探讨不同固定温度和固定时间对免疫组织化学(IHC)染色肝组织中广谱细胞角蛋白(CKpan)有效阳性表达的影响。方法新鲜肝组织标本切割成1.0cm×1.0cm×0.3cm的组织块。在4、25、45℃条件下,分别使用4%中性甲醛固定1、3、9、12、24、48、72h,脱水制备成石蜡切片并进行IHC染色,比较不同固定温度和固定时间的CKpan有效阳性率。结果4℃条件下固定1、3、9、12、24、48、72h的CKpan有效阳性率分别为38.95%、75.79%、86.32%、90.53%、92.63%、95.79%、97.89%(χ^(2)=167.37,P<0.01),25℃条件下固定1、3、9、12、24、48、72h的CKpan有效阳性率分别为70.53%、95.79%、96.84%、98.95%、97.89%、97.89%、98.95%(χ^(2)=105.32,P<0.01),45℃条件下固定1、3、9、12、24、48、72h的CKpan有效阳性率分别为90.53%、92.63%、75.79%、63.16%、54.74%、25.26%、16.84%(χ^(2)=211.31,P<0.01)。4、25℃条件下固定24、48、72h的CKpan有效阳性率均高于45℃条件下(P<0.01)。结论不同固定温度和固定时间对IHC染色肝组织中CKpan有效阳性表达有影响。为提高IHC染色质量,降低固定温度则需延长固定时间,提高固定温度则需缩短固定时间。
Objective To explore the effect of different fixed temperature and fixed time on effective positive expression of pan-cytokeratin(CKpan)in immunohistochemistry(IHC)stained liver tissues.Methods Fresh liver tissue specimens were cut into 1.0cm×1.0cm×0.3cm tissue blocks.At the fixed temperatures of 4,25 and 45℃,4%neutral formaldehyde was used to fix them for 1,3,9,12,24,48 and 72hours,respectively,and paraffin sections were prepared by dehydration and dyed,so as to IHC staining.The effective positive rates of CKpan at different fixed temperatures and fixed times were compared.Results The effective positive rates of CKpan fixed at 4℃for 1,3,9,12,24,48 and 72 hours were 38.95%,75.79%,86.32%,90.53%,92.63%,95.79%and 97.89%,respectively(χ~2=167.37,P<0.01).The effective positive rates of CKpan fixed at 25℃for 1,3,9,12,24,48 and 72 hours were 70.53%,95.79%,96.84%,98.95%,97.89%,97.89%and 98.95%,respectively(χ~2=105.32,P<0.01).The effective positive rates of CKpan fixed at 45℃for 1,3,9,12,24,48 and 72 hours were 90.53%,92.63%,75.79%,63.16%,54.74%,25.26%and 16.84%,respectively(χ~2=211.31,P<0.01).The positive rates of CKpan fixed for 24,48 and 72 hours at 4 and 25℃were higher than those at 45℃(P<0.01).Conclusion Different fixed temperatures and fixed times have an effect on the effective positive expression of CKpan in IHC stained liver tissues.In order to improve the quality of IHC staining,the fixed time should be extended if the fixed temperature is decreased,and the fixed time should be shortened if the fixed temperature is raised.
作者
付裕
韩雪
卓帅帅
杨春青
宋国新
FU Yu;HAN Xue;ZHUO Shuaishuai;YANG Chunqing;SONG Guoxin(Department of Pathology,First Affiliated Hospital,Nanjing Medical University,Nanjing 210029,CHINA)
出处
《江苏医药》
2025年第3期268-271,F0003,共5页
Jiangsu Medical Journal
