本文選題：免疫組織化學　切入點：抗原修復　出處：《大連醫(yī)科大學》2017年碩士論文　論文類型：學位論文

【摘要】：目的:探討免疫組織化學的前處理過程對組織抗原性的影響,尋找影響抗原性的因素并定量分析用于組織處理的試劑對抗原性造成的影響。方法:選取確診為乳腺癌且雌激素受體免疫組化染色為強陽性的組織分別做冰凍切片和石蠟切片樣本,用此樣本做免疫組化染色,用磷酸鹽緩沖液代替一抗作為陰性對照,最后對染色結果進行統(tǒng)計學分析;用乙肝表面抗原陽性的血清標本來模擬組織標本的組織處理過程,于血清中分別加入10%中性福爾馬林、無水酒精、二甲苯、石蠟(模擬石蠟的熔點溫度)、二甲苯及無水酒精,加生理鹽水作對照組;處理后應用抗體吸收試驗法待反應充分后測定其乙肝表面抗體濃度;所得數據用統(tǒng)計學軟件進行統(tǒng)計學分析。結果:冰凍切片和經過抗原修復的石蠟切片的染色結果之間的差異無統(tǒng)計學意義,未經抗原修復的石蠟切片免疫組化染色結果均呈陰性,與前兩者之間的差異有統(tǒng)計學意義;用10%中性福爾馬林處理過的血清與對照組比較其抗原性無差異,而經過無水酒精處理的血清與對照組比較其抗原性有差異(P0.05),在未采用抗原修復處理的情況下其抗原活性損失90%。結論:本研究發(fā)現免疫組織化學的前處理過程中無水酒精試劑對抗原性的影響最大。因此免疫組織化學技術可能只適用于組織中含量較大且抗原性較強抗原的檢測。
[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.
【學位授予單位】：大連醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R361.2

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