目的 探讨抗原修复时间及修复液pH值对Pax-8和CD10免疫组化染色效果的影响。方法 选取肾脏、肝脏手术标本各15份（15例）,采用免疫组化EnVision二步法对肾组织Pax-8和肝组织CD10分别进行pH8.0和pH9.0抗原修复,修复时间为1.5 min、2 min、2.5min及3 min,染色后由病理医师进行盲法阅片及评价。结果 Pax-8和CD10在两组修复液中细胞阳性表达均良好（P<0.05）。肾组织Pax-8抗体染色经双因素方差分析显示修复液pH值主效应显著（P<0.01,偏η²=0.424）,pH8.0修复液修复1.5 min、2 min、2.5 min及3 min组背景染色均浅于pH值9.0各组,差异具有统计学意义（P<0.01）,修复时间及交互效应差异均无统计学意义。肝组织CD10抗体染色经双因素方差分析显示修复液pH主效应显著（P<0.01,偏η²=0.076）,修复时间主效应不显著（P>0.05）,而两者交互效应显著（P<0.05,偏η²=0.068）,经pH8.0修复液修复1.5 min组背景着色显著浅于pH9.0修复1.5 min组（P<0.01）,pH值8.0修复液修复组各时间点间背景着色无统计学差异（P>0.05）,经pH9.0修复液修复时,随着修复时间的延长,染色背景逐渐降低,3 min组背景最浅（P<0.05）。结论 肾组织Pax-8染色背景与抗原修复时间无关,pH8.0修复液修复可降低染色背景。随着修复液pH值的变化,肝组织CD10染色时修复时间应进行适当调整,pH8.0和pH9.0的修复时间分别宜为1.5 min和3 min。
Abstract
Objective To investigate the effect of antigen retrieval time and pH value of the retrieval solution on the immunohistochemical staining results of Pax-8 and CD10. Methods Fifteen surgical specimens of kidneys and livers （15 cases respectively） were selected,and the EnVision two-step immunohistochemical method was used to perform antigen retrieval with pH8.0 and pH9.0 in renal tissue Pax-8 and liver tissue CD10, respectively. The retrieval time was 1.5 min,2 min,2.5 min,and 3 min. The staining was evaluated by the main pathologist in a blind review. Results The positive expression of Pax-8 and CD10 in both groups of retrieval solutions was good（P< 0.05）. The double-factor analysis of variance of renal tissue Pax-8 antibody staining showed that the main effect of retrieval solution pH was significant（P< 0.01, partial η²= 0.424）. Backgroundstaining in pH 8.0 retrieval solution groups of 1.5 min, 2 min, 2.5 min, and 3 min was significantly lighter than that in pH 9.0 groups,with statistically significant difference （P< 0.01）. There was no statistically significant difference in the interaction effect of the retrieval time and the interaction effect between the two factors. For hepatic tissue CD10 antibody staining,double-factor ANOVA indicated a significant main effect of pH （P< 0.01, partial η²= 0.076）, non-significant main effect of retrieval r time （P> 0.05）, but significant interaction effect （P< 0.05, partial η²= 0.068）. The pH 8.0-1.5 min group showed significantly lighter background staining than pH 9.0-1.5 min group （P< 0.01）. Within pH 8.0 groups,no significant differences were observed across time points （P> 0.05）. Under pH 9.0 conditions, background staining progressively decreased with extended repair time, reaching the lightest level in 3-min group. Conclusions The background of Pax-8 staining n renal tissue is not related to antigen retrieval time,while pH 8.0 retrieval solution can reduce the background staining. For hepatic tissue CD10 staining,antigen retrieval duration should be appropriately adjusted according to pH variation of retrieval solutions：1.5 min for pH 8.0 and 3 min for pH 9.0.
关键词
免疫组化 /
抗原修复 /
pH /
背景着色
Key words
immunohistochemistry /
antigen retrieval /
pH /
background coloration
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