Evaluation of the value of frozen tissue section used as "gold standard" for immunohistochemistry.

Author(s): Shi SR, Liu C, Pootrakul L, Tang L, Young A, Chen R, Cote RJ, Taylor CR

Publication: Am J Clin Pathol, 2008, Vol. 129, Page 358

PubMed ID: 18285257 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   This study conducted a comparative assessment of fixation methods (acetone, ethanol, neutral buffered formalin (NBF)) and durations (10-30 min) of frozen tissue sections immediately before immunohistochemistry for 26 antigens. Results were also compared to those obtained with formalin-fixed, paraffin-embedded (FFPE) tissue sections. Experiments were conducted with breast, colon, adrenal gland, and bladder cancers, melanoma, and lymph node tissue.

   Summary of Findings:

   FFPE tissue sections subjected to antigen retrieval yielded superior morphology and immunostaining for the majority of antigens compared to fixed frozen OCT-embedded tissue sections. Among differentially fixed frozen sections, NBF yielded superior morphology and reduced nonspecific staining when coupled with antigen retrieval. Although results were antigen-specific, similar immunostaining intensity was observed among acetone and NBF, or acetone and ethanol fixed sections. While immunostaining was more intense in NBF fixed frozen sections for 31% of the antibodies, no differences were observed with or without the addition of calcium chloride. Antigenicity of cytoplasmic antigens was comparable among acetone and NBF fixed frozen sections, while immunostaining for nuclear proteins and cell surface proteins was superior with NBF and acetone fixation, respectively.

   Biospecimens

   • Tissue - Breast
   • Tissue - Lymph Node
   • Tissue - Bladder
   • Tissue - Skin
   • Tissue - Colorectal
   • Tissue - Adrenal Gland

   Preservative Types

   • Formalin
   • Other Preservative
   • Ethanol

   Diagnoses:

   • Neoplastic - Carcinoma
   • Normal
   • Neoplastic - Melanoma

   Platform:

   Analyte	Technology Platform
   Protein	Immunohistochemistry
   Morphology	Subcellular localization
   Morphology	Light microscopy

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Preservation	Type of fixation/preservation	Acetone Ethanol Neutral-buffered formalin Neutral-buffered formalin + CaCl
   Storage	Storage temperature	-20 degrees C Room temperature
   Biospecimen Preservation	Time in fixative	10 min 30 min Overnight
   Immunohistochemistry Specific	Targeted peptide/protein	Nuclear proteins Cytoplasmic proteins Cell-surface proteins

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