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目的探讨不同抗原修复方法在陈旧性石蜡切片乳腺珠蛋白免疫组化中的效果。方法选取2010年1月至2012年4月本实验室保存的陈旧组织石蜡切片样品76例,所有样品组织均取自乳腺癌患者,各取连续3张切片分别归入试验组、高压组和微波组。微波组和高压组按照常规加热修复法处理相应患者石蜡切片,试验组在pH为3.5±0.1的柠檬酸盐缓冲液中室温修复15min。比较三组切片的抗原修复效果。结果三组切片经过不同的处理方法修复后,肿瘤细胞染色结果显示,试验组染色强度及高阳性比例分布优于高压组和微波组,差异具有统计学意义(χ~2=11.122、14.468、12.859、13.267,P<0.05)。综合阳性标记分数显示,试验组阳性分数多为++、+++,与高压组、微波组分布比较差异有统计学意义(χ~2=13.713、10.331,P<0.01)。试验组阳性率明显高于后两组,差异具有统计学意义(χ~2=6.199、4.475,P<0.05)。试验组经修复后的脱片率(χ~2=22.800、27.059,P<0.01)及折损率(χ~2=64.083、40.827,P<0.01)均明显低于高压组、微波组。结论常温下pH为3.5±0.1柠檬酸盐缓冲液进行抗原修复,能减少组织脱片及折损的发生率,提高切片修复效果。
Objective To investigate the effect of different antigen retrieval methods on the immunohistochemistry of old paraffin sections of mammaglobin. Methods From January 2010 to April 2012, 76 samples of paraffin sections of old tissue preserved in our laboratory were collected. All samples were taken from breast cancer patients. Three consecutive sections were divided into test group, high pressure group and microwave group. In the microwave group and the high-pressure group, paraffin sections of the corresponding patients were treated according to the conventional heat-repair method. The test group was repaired at pH 3.5 ± 0.1 in citrate buffer for 15 minutes at room temperature. The antigenic repair effects of three groups of sections were compared. Results After the three groups of sections were repaired by different treatment methods, the staining results of tumor cells showed that the staining intensity and the positive ratio distribution of the experimental group were better than those of the high-pressure group and the microwave group (χ ~ 2 = 11.122, 14.468, 12.859 , 13.267, P <0.05). The positive labeling score showed that the positive scores in the test group were mostly ++ and +++, which were significantly different from those in the high-pressure group and the microwave group (χ ~ 2 = 13.713,10.331, P <0.01). The positive rate of the experimental group was significantly higher than the latter two groups, the difference was statistically significant (χ ~ 2 = 6.199,4.475, P <0.05). The rate of deprotonation (χ ~ 2 = 22.800, 27.059, P <0.01) and the rate of damage (χ ~ 2 = 64.083, 40.827, P <0.01) in the experimental group were significantly lower than those in the high pressure group and the microwave group. Conclusion The pH value of 3.5 ± 0.1 citrate buffer for antigen retrieval at room temperature can reduce the incidence of tissue delamination and loss and improve the effect of section repair.