Cancer Diagnosis Program | Biorepositories and Biospecimen Research Branch

Search BRD Papers

Influence of fixation and decalcification on the immunohistochemical staining of cell-specific markers in paraffin-embedded human bone biopsies.

Author(s): Mullink H, Henzen-Logmans SC, Tadema TM, Mol JJ, Meijer CJ

Publication: J Histochem Cytochem, 1985, Vol. 33, Page 1103-9

PubMed ID: 2414361 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to determine the effects of fixation and decalcification solutions on IHC staining of bone marrow specimens. In addition, the optimum trypsin concentration for antigen unmasking was determined.

   Summary of Findings:

   A comparison between fixation with 4% neutral buffered formaldehyde and formol sublimate revealed superior immunostaining for most antigens when tissue was fixed in formol sublimate. Trephine biopsies fixed with formol sublimate and decalcified with acetic acid-formol saline generally showed better immunostaining than acetic acid-formol sublimate-fixed aspirates. In general, trypsin concentrations of 0.1-0.5% gave the best balance between background and staining intensity, but results varied based on decalcification solution and antigen. Trypsinization was not necessary for immunostaining of formol sublimate-fixed, acetic acid-formol saline-decalcified specimens.

   Biospecimens

   • Tissue - Bone Marrow

   Preservative Types

   • Other Preservative

   Diagnoses:

   • Autopsy
   • Other diagnoses

   Platform:

   Analyte	Technology Platform
   Protein	Immunohistochemistry

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Preservation	Type of fixation/preservation	Formaldehyde Formol sublimate Acetic acid-formol sublimate
   Preaquisition	Diagnosis/ patient condition	Hematologic disorder Lymphoproliferative disorder
   Immunohistochemistry Specific	Targeted peptide/protein	Immunoglobulin light chain Immunoglobulin heavy chain Elastase Lysozyme Alpha-1-antitrypsin Alpha-1-antichymotrypsin Leukocyte common antigen Hemoglobin Glycophorin A Factor VIII-related antigen CEA 115D8 Keratin
   Analyte Extraction and Purification	Antigen retrieval	None 0.01% trypsin 0.1% trypsin 0.5% trypsin 1% trypsin
   Analyte Extraction and Purification	Decalcification solution/ duration	25% Na2EDTA 10% formic acid 10% acetic acid Kristensens fluid Acetic acid-formol saline Formic acid-formaldehyde saline
   Biospecimen Acquisition	Method of tissue acquisition	Biopsy Aspirate

   View more