Vosse Bettine A H, Seelentag Walter, Bachmann Astrid, Bosman Fred T, Yan Pu
Faculty of Medicine, University of Maastricht, Maastricht, The Netherlands.
Appl Immunohistochem Mol Morphol. 2007 Mar;15(1):103-7. doi: 10.1097/01.pai.0000213102.33816.13.
The aim of this study was to evaluate specific immunostaining and background staining in formalin-fixed, paraffin-embedded human tissues with the 2 most frequently used immunohistochemical detection systems, Avidin-Biotin-Peroxidase (ABC) and EnVision+. A series of fixed tissues, including breast, colon, kidney, larynx, liver, lung, ovary, pancreas, prostate, stomach, and tonsil, was used in the study. Three monoclonal antibodies, 1 against a nuclear antigen (Ki-67), 1 against a cytoplasmic antigen (cytokeratin), and 1 against a cytoplasmic and membrane-associated antigen and a polyclonal antibody against a nuclear and cytoplasmic antigen (S-100) were selected for these studies. When the ABC system was applied, immunostaining was performed with and without blocking of endogenous avidin-binding activity. The intensity of specific immunostaining and the percentage of stained cells were comparable for the 2 detection systems. The use of ABC caused widespread cytoplasmic and rare nuclear background staining in a variety of normal and tumor cells. A very strong background staining was observed in colon, gastric mucosa, liver, and kidney. Blocking avidin-binding capacity reduced background staining, but complete blocking was difficult to attain. With the EnVision+ system no background staining occurred. Given the efficiency of the detection, equal for both systems or higher with EnVision+, and the significant background problem with ABC, we advocate the routine use of the EnVision+ system.
本研究的目的是使用两种最常用的免疫组织化学检测系统，即抗生物素蛋白-生物素-过氧化物酶（ABC）和EnVision+，评估福尔马林固定、石蜡包埋的人体组织中的特异性免疫染色和背景染色。该研究使用了一系列固定组织，包括乳腺、结肠、肾脏、喉、肝脏、肺、卵巢、胰腺、前列腺、胃和扁桃体。选择了三种单克隆抗体，一种针对核抗原（Ki-67），一种针对细胞质抗原（细胞角蛋白），一种针对细胞质和膜相关抗原，以及一种针对核和细胞质抗原的多克隆抗体（S-100）用于这些研究。当应用ABC系统时，在有和没有阻断内源性抗生物素蛋白结合活性的情况下进行免疫染色。两种检测系统的特异性免疫染色强度和染色细胞百分比相当。使用ABC在各种正常细胞和肿瘤细胞中导致广泛的细胞质和罕见的核背景染色。在结肠、胃黏膜、肝脏和肾脏中观察到非常强烈的背景染色。阻断抗生物素蛋白结合能力可减少背景染色，但难以实现完全阻断。使用EnVision+系统未出现背景染色。鉴于两种系统检测效率相同或EnVision+更高，且ABC存在明显的背景问题，我们提倡常规使用EnVision+系统。
