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Abstract：
Spatial relations between tumor cells and host-infiltrating cells are increasingly important in both basic science and clinical research. In this study, we have tested the feasibility of using standard methods of immunohistochemistry (IHC) in a multiplex staining system using a newly developed set of chromogenic substrates for the peroxidase and alkaline phosphatase enzymes. Using this approach, we have developed a set of chromogens characterized by (1) providing fine cellular detail, (2) non-overlapping spectral profiles, (3) an absence of interactions between chromogens, (4) stability when stored, and (5) compatibility with current standard immunohistochemistry practices. When viewed microscopically under brightfield illumination, the chromogens yielded the following colors: red, black, blue, yellow, brown, and green. By selecting compatible color combinations, we have shown feasibility for four-color multiplex staining. Depending on the particular type of analysis being performed, visual analysis, without the aid of computer-assisted image analysis, was sufficient to differentiate up to four different markers.
摘要：
肿瘤细胞和宿主浸润细胞之间的空间关系在基础科学和临床研究中越来越重要。在这项研究中,我们已经测试了在多重染色系统中使用标准免疫组织化学（IHC）方法的可行性，该方法使用了一组新开发的过氧化物酶和碱性磷酸酶的显色底物。使用这种方法，我们开发了一套色原，其特征是(1)提供精细的细胞细节，(2)非重叠光谱轮廓，（3）色原之间没有相互作用，（4）储存时的稳定性，和(5)与当前标准免疫组织化学实践的相容性。在明场照明下显微观察时，色原产生以下颜色：红色，黑色,蓝色,黄色,棕色,和绿色。通过选择兼容的颜色组合，我们已经证明了四色多重染色的可行性。根据所执行的特定分析类型，可视化分析,没有计算机辅助图像分析的帮助，足以区分多达四种不同的标记。
