Utsunomiya Y
Department of Pediatrics, Faculty of Medicine, Tottori University, Yonago, Japan.
Nihon Jinzo Gakkai Shi. 1991 Oct;33(10):949-59.
We devised a periodic acid thionine schiff (PATS)-chromotrope method to detect the glomerular deposits more distinctly than conventional staining methods. The PATS-chromotrope method was compared with other immunological staining methods, such as immunofluorescence method and avidin biotin complex method. Formalin-fixed, and paraffin-embedded renal tissues were obtained from 26 patients with IgA nephropathy, 8 with lupus nephritis, 4 with minimal change nephrotic syndrome, 3 with membrano-proliferative glomerulonephritis, and 3 with hepatitis-B associated nephropathy. Thionine Schiff reagent was used instead of fuchsin-schiff reagent to stain the basement membrane blue. Subsequently, chromotrope 2R was used to stain the glomerular deposits. For immunofluorescence method, frozen renal tissues were stained with FITC-labelled anti-human IgG, IgA, C3 and fibrinogen. For avidin biotin complex method, the same sections as PATS-chromotrope method were stained with anti-human IgG, IgA, and C3. In PATS-chromotrope method, deposits were identified in 9 of 26 specimens with IgA nephropathy, 3 of 8 specimens with lupus nephritis, and one of 3 specimens with hepatitis-B associated nephropathy. Localization of deposits in PATS-chromotrope method was identified more distinctly than immunofluorescence method or avidin biotin complex method. PATS-chromotrope method is useful to detect the deposition of immune complex on routine light microscopy in human glomerular disease.
我们设计了一种过碘酸硫堇席夫(PATS)-变色酸法,以比传统染色方法更清晰地检测肾小球沉积物。将PATS-变色酸法与其他免疫染色方法进行比较,如免疫荧光法和抗生物素蛋白-生物素复合物法。从26例IgA肾病患者、8例狼疮性肾炎患者、4例微小病变肾病综合征患者、3例膜增生性肾小球肾炎患者和3例乙型肝炎相关性肾病患者中获取福尔马林固定、石蜡包埋的肾组织。使用硫堇席夫试剂代替品红-席夫试剂将基底膜染成蓝色。随后,用变色酸2R对肾小球沉积物进行染色。对于免疫荧光法,用异硫氰酸荧光素标记的抗人IgG、IgA、C3和纤维蛋白原对冷冻肾组织进行染色。对于抗生物素蛋白-生物素复合物法,与PATS-变色酸法相同的切片用抗人IgG、IgA和C3进行染色。在PATS-变色酸法中,26例IgA肾病标本中有9例、8例狼疮性肾炎标本中有3例以及3例乙型肝炎相关性肾病标本中有1例检测到沉积物。PATS-变色酸法中沉积物的定位比免疫荧光法或抗生物素蛋白-生物素复合物法更清晰。PATS-变色酸法有助于在人类肾小球疾病的常规光学显微镜检查中检测免疫复合物的沉积。