石蜡切片免疫荧光染色在肾活检组织病理诊断中的应用

Application of immunofluorescence staining of paraffin embedded human renal biopsy samples in pathological diagnosis

目的探讨使用10%中性福尔马林固定、石蜡包埋的肾活检组织做免疫荧光染色在病理诊断应用中的可行性。方法选择慢性肾脏病患者的肾穿刺活检组织标本,分别留取作石蜡切片及冰冻切片。对石蜡切片进行脱蜡、抗原修复及直接免疫荧光染色,并将染色结果与冰冻切片免疫荧光染色结果进行比较。结果石蜡切片经高温修复联合酶消化法进行免疫荧光染色,C1q及C3阳性率、分布部位和强度与冰冻切片免疫荧光染色结果相一致;石蜡切片IgA、IgM及IgG染色分布部位与冰冻切片无差别,阳性率和染色强度略低于冰冻切片,但不影响诊断,其结果与冰冻切片染色的诊断符合率吻合度较高。且石蜡切片具有组织结构保存好、免疫沉积物清晰、易于判断结果等优点。结论石蜡切片与冰冻切片的免疫荧光染色效果相当,可在肾脏病理诊断中作为冰冻切片免疫荧光染色一种较好的技术补充方法。

Objective To investigate the feasibility of immunofluoreseent staining （IF） in formalin-fixed and paraffin-embedded human renal tissue. Methods Renal biopsy tissue samples from the patients with chronic kidney diseases were obtained for making paraffin sections and frozen sections. The paraffin sections were treated by deparaffinization, antigen retrieval and direct IF, and the results were compared. Results Paraffin-embedded sections of renal biopsy tissues were processed by heat antigen retrieval combined with pancreatin digestion and immunofluorescence staining of IgA, IgM,IgG,C3 and Clq. The staining of Clq and C3 was in accordance with that of frozen sections in positive rate, distribution pattern and staining intensity. The IgA, IgM, IgG were the same with those by IF of frozen section in distribution pattern, but the intensity and positive rate were lower than those by frozen section, which did not influence diagnosis. Furthermore, the structure of paraffin sections was kept better and clearer than that of frozen sections, which led to the diagnosis easier. Conclusion The IF on paraffin embedded section may become a complementary method of IF on frozen section in renal pathology diagnosis.