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《Chinese Journal of Integrated Traditional and Western Nephrology》 2008-07
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The Application of Microwave-assisted Multicolor Fluorescence Iimmunos-taining in Paraffin-embedded Renal Biopsy Tissues

SHI Suozhu,CHEN Xiangmei,CHEN Pu,et al Department of Nephrology,General Hospital of Chinese PLA,Beijing(100853)  
Objective:to explore the practicality of multiple stainig with immunofluorescence in formalin-fixed and paraffin-embedded human renal tissue slices.Methods:Microwave technology permitted the development of new antigen labeling techniques.The present study tested multicolor labeling of three antibodies in routinely processed paraffin-embedded renal biopsy tissue sections using a microwave-assisted immunofluorescence technique.Biopsy samples from patients with IgA nephropathy(n=5),lupus nephritis(n=5),membranous nephropathy(n=5)and HBV related nephritis(n=5)were taken for simultaneous staining of IgG/Fn/Hoechst,C3/Fn/Hoechst,IgA/Fn/Hoechst and HBsAg/Fn/Hoechst.Antigen unmasking was performed with microwave treatment.The paraffin sections and frozen sections were analyzed by fluorescence microscope.Results:The multicolor fluorescence immunostaining of microwave-assisted paraffin sections showed that the stainings of IgG,C3,IgA and HBsAg were green,and FN was red.The coexpression of two different antigens was orange with blue nucleus.The results were in accordance with that of frozen sections.Furthermore,the paraffin sections were thinner and clearer,which lead to more specific location of immune deposits and easier judgment.Conclusion:The application of microwave-assisted immunofluorescence technique can show more than two antigens simultaneously in one paraffin section.This method is effective for revealing more accurate and direct details and useful for immune pathologic diagnosis of renal biopsy tissue.
【Fund】: 国家自然科学基金资助项目(No.30630033);; 国家自然科学基金面上资助项目(No.30570856)
【CateGory Index】: R446.8
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