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A technique for retrieving antigens in formalin-fixed, routinely acid-decalcified, celloidin-embedded human temporal bone sections for immunohistochemistry.

Author(s): Shi SR, Coté C, Kalra KL, Taylor CR, Tandon AK

Publication: J Histochem Cytochem, 1992, Vol. 40, Page 787-92

PubMed ID: 1588025 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to validate a novel antigen retrieval technique for immunohistochemical staining of formalin or Heidenhain-Susa fixed, decalcified, celloidin-embedded human temporal bone sections. The monoclonal antibodies tested were specific to keratin AE1, keratin NCL-5D3, vimentin, neurofilament, glial fibrillary acidic protein, desmin, myoglobin, alpha-tubulin, beta-tubulin, muscle-specific actin, chromogranin, alpha actinin, epithelial membrane antigen, neuron-specific enolase, and S-100 protein.

   Summary of Findings:

   The authors report that a strong alkaline solution was key to successful antigen retrieval. Methanol saturated with NaOH performed best, as ethanol saturated with NaOH yielded less intense positive staining. Of the 15 antibodies evaluated, strong, moderate, and weak immunostaining was observed for 7, 4, and 1 antibodies, respectively. Although comparisons to alternative antigen retrieval protocols were limited, the authors noted the absence of staining in negative controls and predicted cell-specific immunostaining after a 30 min incubation in methanol saturated with NaOH for the following antigens: neurofilament, neuron-specific enolase, keratin, actin, and vimentin. Significant differences between sections from different blocks were not observed, regardless of the fixative or decalcification method employed, or storage duration.

   Biospecimens

   • Tissue - Bone

   Preservative Types

   • Other Preservative
   • Formalin

   Diagnoses:

   • Not specified

   Platform:

   Analyte	Technology Platform
   Protein	Immunohistochemistry

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Analyte Extraction and Purification	Antigen retrieval	Methanol saturated with NaOH Ethanol saturated with NaOH
   Biospecimen Preservation	Type of fixation/preservation	Formalin (buffered) Heidenhain-Susa solution Formalin (unbuffered)
   Analyte Extraction and Purification	Decalcification solution/ duration	Ethylenediaminetetraacetic acid (EDTA) Trichloroacetic acid
   Storage	Storage duration	<1 yr <2 yr 4 yr 8 yr 30 yr

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