Fogazzi G B, Bajetta M, Banfi G, Mihatsch M
Division of Nephrology and Dialysis, Maggiore Hospital, Milan, Italy.
Pathol Res Pract. 1989 Aug;185(2):225-30. doi: 10.1016/S0344-0338(89)80256-0.
Two types of tissue preparation for immunofluorescent staining were compared with sections of the same kidney. In one, formalin-fixed paraffin-embedded kidney sections were incubated with pronase (IF-PRON). The optimal pronase exposure for bright, specific IF and a mild background IF, suitable for the whole set of antisera was determined (pronase 0.75 g/l of Tris buffer for 60 min) and this was used for the subsequent steps of the study. Snap-frozen sections were also stained (IF-FROZ). Positive and negative cases, IF intensity, distribution and location in IgA nephropathy, membranous nephropathy, proliferative lupus nephritis were compared by the 2 methods. The main antigens for each disease were adequately revealed by IF-PRON, so that a correct diagnosis was possible in all cases. IF-PRON was also applied to sections before and after prolonged storage of blocks in the files, to see whether or not retrospective analysis is possible. Only minor differences were found between the 2 series of sections. Finally, the exposure of the sections to the continuous fluorescent light showed that IF fading was less in fixed sections. We conclude that IF-PRON is a reliable method for renal pathology with some advantages over the IF-FROZ.
将两种用于免疫荧光染色的组织制备方法与取自同一肾脏的切片进行了比较。一种方法是,将福尔马林固定石蜡包埋的肾脏切片用链霉蛋白酶处理(IF-PRON)。确定了能产生明亮、特异性免疫荧光且背景免疫荧光较弱的链霉蛋白酶最佳处理条件,该条件适用于整套抗血清(链霉蛋白酶0.75 g/l Tris缓冲液,处理60分钟),并将其用于后续研究步骤。还对速冻切片进行了染色(IF-FROZ)。采用这两种方法比较了IgA肾病、膜性肾病、增殖性狼疮性肾炎中的阳性和阴性病例、免疫荧光强度、分布及定位情况。IF-PRON能充分显示每种疾病的主要抗原,因此在所有病例中均能做出正确诊断。还将IF-PRON应用于切片在档案中长期保存前后,以观察是否可行回顾性分析。两组切片之间仅发现微小差异。最后,将切片暴露于持续荧光下显示,固定切片中的免疫荧光褪色较少。我们得出结论,IF-PRON是一种可靠的肾脏病理学方法,相对于IF-FROZ具有一些优势。