Immunohistochemical detection of p53--effect of fixation and methods of antigen retrieval.

Author(s): Bassarova AV, Popov AA

Publication: Folia Histochem Cytobiol, 1998, Vol. 36, Page 127-32

PubMed ID: 9773296 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   Immunohistochemical detection of p53 was evaluated in breast tumor tissue using four different methods of fixation.

   Summary of Findings:

   After fixation in 70% ethanol or in Carnoy solution, strong staining of p53 was detected in the nuclei and/or cytoplasm of breast tumor cells. Immunopositive staining was relatively weak, hard to detect due to strong background, or absent when tissue was fixed in 10% NBF or in Bouin solution.

   Biospecimens

   • Tissue - Breast

   Preservative Types

   • Ethanol
   • Other Preservative
   • Formalin

   Diagnoses:

   • Neoplastic - Carcinoma
   • Neoplastic - Benign

   Platform:

   Analyte	Technology Platform
   Protein	Immunohistochemistry

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Preservation	Type of fixation/preservation	Bouin's fixative Carnoy's solution Ethanol Formalin (buffered)

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2. Study Purpose

   The purpose of this study was to determine if antigen unmasking was able to improve the quality of immunohistochemical staining of p53 in breast tumor tissue fixed in 10% NBF or in Bouin solution.

   Summary of Findings:

   Antigen retrieval with microwaves yielded increased p53 staining and decreased background on samples fixed in 10% NBF or Bouin solution. Antigen unmasking with 0.1% trypsin resulted in weak or no staining when applied for 10 minutes and high background staining impeding morphological assessment when application time was increased to 20 minutes.

   Biospecimens

   • Tissue - Breast

   Preservative Types

   • Formalin
   • Other Preservative

   Diagnoses:

   • Neoplastic - Carcinoma
   • Neoplastic - Benign

   Platform:

   Analyte	Technology Platform
   Protein	Immunohistochemistry

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Analyte Extraction and Purification	Antigen retrieval	10 mM Citrate buffer 0.1% Trypsin solution 10 min 0.1% Trypsin solution 20 min
   Analyte Extraction and Purification	Temperature of heat-induced retrieval	Room temperature Microwave heating
   Biospecimen Preservation	Type of fixation/preservation	Formalin (buffered) Bouin's fixative

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