Assessment of problems in diagnostic and research immunohistochemistry associated with epitope instability in stored paraffin sections.

Author(s): van den Broek LJ, van de Vijver MJ

Publication: Appl Immunohistochem Mol Morphol, 2000, Vol. 8, Page 316-21

PubMed ID: 11127924 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to determine if the duration and temperature of slide-mounted tissue section storage affects the immunoreactivity of more than 30 antigens. Pituitary glands procured from surgical excision and autopsy were analyzed.

   Summary of Findings:

   Although effects were antigen-specific, the intensity of immunostaining was inversely proportional to storage duration and temperature of slide-mounted tissue sections for several of the antigens evaluated. Specifically, the percentage of stable antigens decreased from 87 to 65 to 50% in sections stored at 4, 21, and 37 degrees C. The maximum storage duration for each antigen was outlined in detail for each storage temperature. Immunostaining intensity was compromised as soon as 1 d after sectioning or remained stable for 360 d. While no differences in immunostaining were observed when monoclonal versus polyclonal antibodies were used with stored versus fresh tissue sections, staining patterns displayed differential results, as membrane staining was more susceptible to prolonged storage durations and elevated temperatures compared to nuclear and cytoplasmic staining. Antigen retrieval (heat- or enzyme-mediated) further impaired immunoreactivity at all storage temperatures.

   Biospecimens

   • Tissue - Pituitary Gland

   Preservative Types

   • Formalin

   Diagnoses:

   • Not specified
   • Autopsy

   Platform:

   Analyte	Technology Platform
   Protein	Immunohistochemistry
   Peptide	Immunohistochemistry

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Storage	Storage duration	0 d 15 d 30 d 60 d 90 d 120 d 180 d 240 d 300 d 360 d
   Storage	Storage temperature	4 degrees C 21 degrees C 37 degrees C
   Analyte Extraction and Purification	Antigen retrieval	Citrate buffer, pH 6 (10 mmol/L) 0.1% Trysin in CaCl 0.4% Pepsin in HCl None

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2. Study Purpose

   The purpose of this study was to determine if the type of adhesive used for tissue section/slide mounting affects immunoreactivity and antigen stability when sections are stored at room temperature. Specimens analyzed were surgically resected tonsil and colon tissue.

   Summary of Findings:

   The adhesives examined did not influence immunohistochemistry results or antigen stability.

   Biospecimens

   • Tissue - Colorectal
   • Tissue - Tonsil

   Preservative Types

   • Formalin

   Diagnoses:

   • Not specified

   Platform:

   Analyte	Technology Platform
   Protein	Immunohistochemistry

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Aliquots and Components	Tissue section adhesion	None Gelatin APES APES and glutaraldehyde Poly-L-lysine
   Storage	Storage duration	1 d 15 d 30 d 45 d 90 d 6 months

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