van der Loos C M, Das P K, Houthoff H J
Department of Pathology, Academic Medical Center, Amsterdam, The Netherlands.
J Histochem Cytochem. 1987 Nov;35(11):1199-204. doi: 10.1177/35.11.2443555.
For immunohistological analysis, simultaneous detection of multiple cellular epitopes, as compared to single staining of serial sections, is sometimes needed. Therefore, immunoenzyme triple-staining protocols were tested with polyclonal and monoclonal antibodies on tissue sections and cytospin preparations. Various immunoconjugates were used in different combinations of methods, of which not all proved to be suitable. Of the tested protocols, one yielded superior results for both monoclonal and polyclonal antibodies, with optimal preservation of their original avidity. The method consists of a combination of indirect, direct, and avidin-biotin complex technique. The three antigens can be distinguished clearly and selectively by the reaction products of the enzyme activities of beta-galactosidase (green), alkaline phosphatase (blue), and horseradish peroxidase (red).
对于免疫组织学分析，与连续切片的单染色相比，有时需要同时检测多个细胞表位。因此，在组织切片和细胞涂片制备物上用多克隆和单克隆抗体对免疫酶三重染色方案进行了测试。各种免疫缀合物以不同的方法组合使用，但并非所有组合都证明是合适的。在测试的方案中，有一种方案对单克隆和多克隆抗体都产生了优异的结果，同时能最佳地保留它们原来的亲和力。该方法由间接法、直接法和抗生物素蛋白-生物素复合物技术组合而成。通过β-半乳糖苷酶（绿色）、碱性磷酸酶（蓝色）和辣根过氧化物酶（红色）的酶活性反应产物，可以清晰且选择性地区分三种抗原。
