肺腺癌肿瘤相关巨噬细胞的免疫组织化学双标染色方法优化

发布时间：2018-04-14 07:31

本文选题：免疫组织化学染色 + 辣根过氧化酶　；参考：《第三军医大学学报》2017年11期

【摘要】：目的通过对现有的免疫组织化学双重标记方法(双标)的改进,探索一种稳定且易于辨认的显色方法,便于在普通光学显微镜下观察肺腺癌组织中肿瘤相关巨噬细胞的分布情况。方法改进的免疫组织化学双标染色分别使用二抗链接的碱性磷酸酶和辣根过氧化物酶作为底物显色酶,底物分别选用固红(AP-Red)和二氨基联苯胺(DAB),在显微镜下次序控制双标显色,观察肺癌组织中肿瘤相关巨噬细胞的分布情况。结果免疫组织化学双重标记中第一标记AP-Red的红色与第二标记DAB的棕黄色反差大,背景低,易于分辨。两种颜色标记的同一部位呈现砖红色,易于表示双标部位。统计显示在肺腺癌组织标本中,以M2型肿瘤相关巨噬细胞为主,约占80%。结论改进后的显色系统建立了简便、可靠的免疫组织化学双重标记新方法。标记结果可以清晰稳定显示肿瘤相关巨噬细胞在肺腺癌组织中分布情况,便于进一步研究其在肺癌发生、发展中的作用。
[Abstract]:Objective to explore a stable and easily recognizable method for color development by improving the existing double labeling method (double labeling) of immunohistochemistry.It is convenient to observe the distribution of tumor-associated macrophages in lung adenocarcinoma tissue under general optical microscope.Methods the modified double labeling immunohistochemical staining was performed by using the linked alkaline phosphatase and horseradish peroxidase as the substrate chromogenic enzyme, and the substrate AP-Red and the diaminobenzidine DABX, respectively. The double standard color was controlled in order under the microscope.The distribution of tumor associated macrophages in lung cancer tissues was observed.Results in the double labeling of immunohistochemistry, the red of the first label AP-Red and the brown yellow of the second label DAB had a big contrast, low background and easy to distinguish.The same part of the two color marks appears brick red, easy to represent the double-marked area.Statistics showed that M 2 tumor associated macrophages were predominant in lung adenocarcinoma tissues, accounting for about 80%.Conclusion the improved color system has established a simple and reliable double labeling method for immunohistochemistry.The results of labeling can clearly and stably show the distribution of tumor associated macrophages in lung adenocarcinoma tissues and facilitate further study of the role of tumor associated macrophages in the genesis and development of lung cancer.
【作者单位】：第三军医大学西南医院全军临床病理学研究所/西南癌症中心;
【基金】：国家重点研发计划重点专项(2016YFC201801) 国家自然科学基金面上项目(81372273) 西南医院生物强院科技创新计划(SWH2016JCZD-04)~~
【分类号】：R734.2

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