Hashizume K, Hatanaka Y, Kamihara Y, Tani Y
Department of Immunohistochemistry, DAKO Japan, Kyoto.
Appl Immunohistochem Mol Morphol. 2001 Mar;9(1):54-60.
An immunohistochemical assay using catalyzed signal amplification (CSA), which is based on the peroxidase catalyzed deposition of biotinylated tyramide, is a highly sensitive method to visualize weak immunohistochemical signals originating from rare antigens or masked antigens in formalin-fixed, paraffin-embedded (FFPE) tissues. However, CSA methods are hampered by poor reproducibility and the complexity of their staining procedures. In this study, we aimed to apply the CSA procedure to a capillary gap-based, automated immunostainer, TechMate Horizon, to perform immunohistochemical signal amplification effectively and reproducibly. A variety of cellular antigens previously considered to be undetectable in FFPE human specimens were selected and examined with the automated immunostainer. Compared with the manual CSA staining method that takes more than 2 hours, the automated CSA method took less than 2 hours to complete. The staining results from the automated CSA method presented higher reproducibility, as well as lower background owing to well-regulated, punctual staining and washing at every step of the procedure. Conclusively, the automation of the CSA method enabled us to perform the time-consuming and complicated CSA amplification technique with minimal effort in an accurate, consistent, and reproducible manner.
基于过氧化物酶催化生物素化酪胺沉积的催化信号放大（CSA）免疫组织化学检测法，是一种用于可视化源自福尔马林固定、石蜡包埋（FFPE）组织中稀有抗原或隐蔽抗原的微弱免疫组织化学信号的高灵敏度方法。然而，CSA方法受到重现性差和染色程序复杂的阻碍。在本研究中，我们旨在将CSA程序应用于基于毛细间隙的自动化免疫染色仪TechMate Horizon，以有效且可重复地进行免疫组织化学信号放大。选择了多种先前认为在FFPE人类标本中无法检测到的细胞抗原，并使用该自动化免疫染色仪进行检测。与耗时超过2小时的手动CSA染色方法相比，自动化CSA方法完成检测所需时间不到2小时。自动化CSA方法的染色结果具有更高的重现性，并且由于在程序的每个步骤中染色和洗涤均得到良好调控、精准到位，背景也更低。总之，CSA方法的自动化使我们能够以准确、一致且可重复的方式，以最小的工作量完成耗时且复杂的CSA放大技术。
