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Immunohistochemistry of deparaffinised sections using antigen retrieval with microwave combined pressure cooking versus immunofluorescence in the assessment of human renal biopsies
  1. Suozhu Shi1,
  2. Ping Zhang2,
  3. Qingli Cheng3,
  4. Jie Wu1,
  5. Jing Cui1,
  6. Ying Zheng1,
  7. Xue-Yuan Bai1,
  8. Xiangmei Chen1
  1. 1Department of Nephrology, State Key Laboratory of Kidney Diseases, Chinese PLA General Hospital, Beijing, China
  2. 2Department of Pathology, Wang Jing Hospital, Chinese Academy of Chinese Medical Sciences, Beijing, China
  3. 3Department of Geriatric Nephrology, Chinese PLA General Hospital, Beijing, China
  1. Correspondence to Professor Xue-Yuan Bai and Xiangmei Chen, Department of Nephrology, State Key Lab of Kidney Diseases, Chinese PLA General Hospital, 28 Fuxing Road, Beijing 100853, China; xueyuan_bai{at}yahoo.com.cn, xmchen301{at}126.com

Abstract

Background Immunofluorescence of frozen tissue sections (IF-F) is a traditional technique used in renal biopsy. However, IF-F has certain disadvantages, such as a few or even no glomeruli in the section, and limited long-term preservation of the fluorescently labelled samples.

Methods We compared two-step immunohistochemistry (IHC) staining of deparaffinised sections for antigen retrieval with microwave combined high-pressure cooking to IF-F used to detect antigens of IgG, IgA, IgM, C3, C1q, κ and λ in patient renal biopsy samples. The number of glomeruli detected, sensitivity and specificity of positive staining, tissue structure, and location staining of the antigens were determined using the two methods in 285 patients diagnosed with different renal diseases.

Results Concordant observations between IF-F and IHC were 99% for all antigen staining (1969 of 1995 observations) and 100% for IgG, IgA and IgM (all 285 observations). The number of glomeruli in IHC sections was significantly greater compared with IF-F sections (p<0.001). IHC provided clearer images of tissue structure, more precise localisation of positive-staining antigens, and IHC staining allowed simultaneous evaluation of tissue by light microscopy. Correlation between tissue structure and immune deposits are not readily attained by IF-F.

Conclusions IHC is superior to IF-F for immunopathological diagnosis of renal biopsy tissue and is a reliable replacement for the more traditional IF-F method.

  • Antigen Retrieval
  • Diagnosis
  • Immunohistochemistry
  • Kidney
  • Microwave

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