Effects of antigen retrieval by microwave heating in formalin-fixed tissue sections on a broad panel of antibodies.

Author(s): von Wasielewski R, Werner M, Nolte M, Wilkens L, Georgii A

Publication: Histochemistry, 1994, Vol. 102, Page 165-72

PubMed ID: 7532634 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to identify an optimal antigen retrieval technique for immunohistochemistry of FFPE normal and cancerous tissue specimens. A total of 52 markers were investigated in nine tissue types.

   Summary of Findings:

   The authors identified boiling slides for 20-35 min in citric acid monohydrate as the superior antigen retrieval protocol, although immunostaining was enhanced for only 9 of the 52 antigens investigated when specimens were fixed for 24 h. Tissue morphology was impaired to some degree, with effects noted in connective tissue and bone marrow biopsies. Heating via microwave or a hot plate produced equivalent results.

   Biospecimens

   • Tissue - Small Bowel
   • Tissue - Bone Marrow
   • Tissue - Tonsil
   • Tissue - Skin
   • Tissue - Kidney
   • Tissue - Lymph Node
   • Tissue - Liver
   • Tissue - Pancreas
   • Tissue - Nerve

   Preservative Types

   • Formalin

   Diagnoses:

   • Neoplastic - Carcinoma
   • Neoplastic - Lymphoma
   • Neoplastic - Melanoma
   • Normal

   Platform:

   Analyte	Technology Platform
   Protein	Immunohistochemistry

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Analyte Extraction and Purification	Antigen retrieval	Citric buffer Bidistilled water Citric acid monohydrate Antigen retrieval solution (BioGenex) No treatment Heating via a microwave Heating via a hot plate
   Biospecimen Acquisition	Biospecimen location	Small intestine Bone marrow Tonsil Skin Nerve Lymph node Liver Pancreas Kidney

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2. Study Purpose

   The aim of this study was to assess the effects of over-fixation (ranging from 1 week to 3 years or longer) on immunohistochemistry of FFPE normal and cancerous tissue specimens. A total of 52 markers were investigated in nine tissue types. Results within author specified groups differing in the duration of fixation were compared with and without antigen retrieval.

   Summary of Findings:

   Antigen retrieval (boiling in citric acid monohydrate) improved immunostaining in tissue specimens fixed in formalin for 1 week or longer, with 50-58% of the 52 antigens evaluated exhibiting enhanced staining after antigen retrieval compared to 17% in specimens fixed for 24 h. False positive staining after antigen retrieval was limited to a single antigen, CD3, and was consistently observed.

   Biospecimens

   • Tissue - Tonsil
   • Tissue - Skin
   • Tissue - Nerve
   • Tissue - Lymph Node
   • Tissue - Bone Marrow
   • Tissue - Pancreas
   • Tissue - Kidney
   • Tissue - Small Bowel
   • Tissue - Liver

   Preservative Types

   • Formalin

   Diagnoses:

   • Normal
   • Neoplastic - Carcinoma
   • Neoplastic - Lymphoma
   • Neoplastic - Melanoma

   Platform:

   Analyte	Technology Platform
   Protein	Immunohistochemistry

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Preservation	Time in fixative	24 h 1-6 weeks 3 y and longer
   Analyte Extraction and Purification	Antigen retrieval	Citric acid monohydrate No treatment

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