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
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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
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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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
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