Yamashina M, Takami T, Kanemura T, Orii T, Ojima A
Department of Pediatrics, Gifu University School of Medicine, Japan.
Lab Invest. 1989 Feb;60(2):311-6.
Although there are many articles describing the detection of complement components in formalin-fixed and paraffin-embedded renal tissues, it is still difficult to obtain reproducible results. The authors determined the optimal conditions for detecting complement components in routine paraffin sections by the avidin-biotin-peroxidase complex method and concluded that proteolytic digestion of the deparaffinized sections was crucial to detect complement and to preserve tissue structures. The optimal proteolytic digestion was a 15-minute incubation at 37 degrees C in trypsin solution, 0.5 mg/ml, pH 7.6. Under these conditions, all of the complement components so far studied (C1q, C1s, C4, C3c, C3b, C5, C6, C9, and properdin) were detected without significant destruction of tissue structures in 52 renal biopsy specimens from patients with various forms of glomerulonephritis. Immunohistochemistry using the avidin-biotin-peroxidase complex method was equivalent to indirect immunofluorescence and superior to direct immunofluorescence in utility and was useful in the diagnosis of glomerular diseases.
虽然有许多文章描述了在福尔马林固定、石蜡包埋的肾组织中检测补体成分，但仍难以获得可重复的结果。作者通过抗生物素蛋白-生物素-过氧化物酶复合物法确定了在常规石蜡切片中检测补体成分的最佳条件，并得出结论，对脱石蜡切片进行蛋白水解消化对于检测补体和保存组织结构至关重要。最佳的蛋白水解消化是在37℃下于0.5mg/ml、pH7.6的胰蛋白酶溶液中孵育15分钟。在这些条件下，在来自各种形式肾小球肾炎患者的52份肾活检标本中，检测到了迄今为止研究的所有补体成分（C1q、C1s、C4、C3c、C3b、C5、C6、C9和备解素），且组织结构未受到明显破坏。使用抗生物素蛋白-生物素-过氧化物酶复合物法的免疫组织化学在实用性上等同于间接免疫荧光，且优于直接免疫荧光，对肾小球疾病的诊断很有用。
