Immunohistochemical staining of plastic embedded bone marrow trephine biopsy specimens after microwave heating.

Author(s): McCluggage WG, Roddy S, Whiteside C, Burton J, McBride H, Maxwell P, Bharucha H

Publication: J Clin Pathol, 1995, Vol. 48, Page 840-4

PubMed ID: 7490318 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 method, embedding material, decalcification, and antigen retrieval on IHC staining and morphological preservation in bone marrow trephine biopsies. Specimens designated for paraffin embedding were fixed in 3% acetic acid in 10% unbuffered formol saline for 1.5-3 h and decalcified with EDTA. Specimens designated for plastic embedding were fixed overnight in 10% neutral buffered formalin and underwent antigen retrieval prior to staining.

   Summary of Findings:

   19 of 22 antibodies gave IHC staining results that were comparable between acetic acid-formol saline-fixed, paraffin-embedded, decalcified bone marrow trephine biopsies and those that were formalin-fixed, embedded in plastic (Polarbed 812 epoxy resin), and treated by microwave heating in citrate buffer prior to immunostaining. CD20 immunostaining was superior in formalin-fixed plastic-embedded specimens, while neutrophil elastase and CD61 immunostaining were superior in acetic acid-formol saline-fixed paraffin-embedded specimens. Cellular morphology was better in formalin-fixed plastic-embedded specimens which were able to be cut into 1 uM sections as opposed to 5 uM sections.

   Biospecimens

   • Tissue - Bone Marrow

   Preservative Types

   • Formalin
   • Other Preservative

   Diagnoses:

   • Not specified
   • Normal
   • Neoplastic - Carcinoma
   • Neoplastic - Lymphoma

   Platform:

   Analyte	Technology Platform
   Protein	Immunohistochemistry
   Morphology	H-and-E microscopy
   Morphology	Light microscopy

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Preservation	Type of fixation/preservation	Acetic acid-formol saline Formalin (buffered)
   Biospecimen Preservation	Embedding medium	Epoxy resin Paraffin
   Analyte Extraction and Purification	Decalcification solution/ duration	EDTA pH 7.0 None
   Analyte Extraction and Purification	Antigen retrieval	Microwaved in citrate buffer None
   Biospecimen Aliquots and Components	Type of slide	1 uM section 5 uM section

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