免疫组织化学技术关键步骤对组织抗原影响的定量分析

发布时间：2018-03-21 02:19

本文选题：免疫组织化学　切入点：抗原修复　出处：《大连医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:探讨免疫组织化学的前处理过程对组织抗原性的影响,寻找影响抗原性的因素并定量分析用于组织处理的试剂对抗原性造成的影响。方法:选取确诊为乳腺癌且雌激素受体免疫组化染色为强阳性的组织分别做冰冻切片和石蜡切片样本,用此样本做免疫组化染色,用磷酸盐缓冲液代替一抗作为阴性对照,最后对染色结果进行统计学分析;用乙肝表面抗原阳性的血清标本来模拟组织标本的组织处理过程,于血清中分别加入10%中性福尔马林、无水酒精、二甲苯、石蜡(模拟石蜡的熔点温度)、二甲苯及无水酒精,加生理盐水作对照组;处理后应用抗体吸收试验法待反应充分后测定其乙肝表面抗体浓度;所得数据用统计学软件进行统计学分析。结果:冰冻切片和经过抗原修复的石蜡切片的染色结果之间的差异无统计学意义,未经抗原修复的石蜡切片免疫组化染色结果均呈阴性,与前两者之间的差异有统计学意义;用10%中性福尔马林处理过的血清与对照组比较其抗原性无差异,而经过无水酒精处理的血清与对照组比较其抗原性有差异(P0.05),在未采用抗原修复处理的情况下其抗原活性损失90%。结论:本研究发现免疫组织化学的前处理过程中无水酒精试剂对抗原性的影响最大。因此免疫组织化学技术可能只适用于组织中含量较大且抗原性较强抗原的检测。
[Abstract]:Objective: to investigate the effect of the pretreatment process of immunohistochemistry on tissue antigenicity. To find the factors affecting antigenicity and quantitatively analyze the effect of the reagent used in tissue processing on antigenicity. Methods: the tissues diagnosed with breast cancer and strongly positive for estrogen receptor immunocytochemical staining were frozen respectively. Samples of slices and paraffin sections, This sample was used for immunohistochemical staining and phosphate buffer solution was used instead of the first antibody as negative control. Finally, the staining results were statistically analyzed, and the tissue processing process of tissue samples was simulated by using serum samples positive for hepatitis B surface antigen. 10% neutral formalin, anhydrous alcohol, xylene, paraffin (temperature of melting point of simulated paraffin), xylene and anhydrous alcohol were added to serum, and normal saline was added as control group. After treatment, the antibody concentration on hepatitis B surface was determined by antibody absorption test. Results: there was no significant difference between the staining results of frozen sections and paraffin sections repaired with antigen. The results of immunohistochemical staining of paraffin sections without antigen repair were all negative, and the difference was statistically significant between the two groups, and the antigenicity of serum treated with 10% neutral formalin was not different from that of the control group. The antigenicity of serum treated with anhydrous alcohol was different from that of control group (P 0.05). The antigenic activity of serum treated with anhydrous alcohol was 90% lower than that of control group. Conclusion: this study found that there was no antigenicity in the pretreatment of immunohistochemistry. Water alcohol reagents have the greatest influence on antigenicity, so the immunohistochemical technique may only be suitable for the detection of high content and strong antigenicity in tissues.
【学位授予单位】：大连医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R361.2

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