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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。
OBJECTIVE To explore a stable and easily identifiable chromogenic method by improving the existing immunohistochemical double labeling method (double labeling) and to facilitate the observation of the distribution of tumor-associated macrophages in lung adenocarcinoma tissues under an ordinary light microscope Happening. Methods Improved immunohistochemical double-stained staining using secondary antibody linked to alkaline phosphatase and horseradish peroxidase as substrate chromogenic enzymes were selected substrate red-AP (Red) and diaminobenzidine ( DAB) under the microscope in order to control the double standard color, observe the distribution of tumor-associated macrophages in lung cancer. Results The red of the first marker AP-Red in the immunohistochemical double marker was larger than that of the second marker DAB in brown background, and the background was low and easy to distinguish. The same part of the two color markers appear brick red, easy to represent double-standard parts. Statistics show that in lung adenocarcinoma tissue samples to M2 tumor-associated macrophages, accounting for about 80%. Conclusion The improved colorimetric system has established a simple and reliable new immunohistochemical double labeling method. The labeling results can clearly and stably display the distribution of tumor-associated macrophages in lung adenocarcinoma tissue for further study of its role in the occurrence and development of lung cancer.