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Related Experiment Videos

Antigenicity testing by immunohistochemistry after tissue oxidation.

C Blind1, A Koepenik, M Pacyna-Gengelbach

  • 1Institute of Pathology, Charité Campus Mitte, Charitéplatz 1, 10117 Berlin.

Journal of Clinical Pathology
|April 7, 2007
PubMed
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Photo-oxidation and drying degrade tissue antigenicity over time, impacting immunohistochemistry results. Protocols are provided to test antigen stability under these conditions.

Area of Science:

  • Histopathology
  • Immunohistochemistry
  • Biomarker Stability

Background:

  • Archived tissue blocks generally maintain antigenicity, but tissue sections can lose immunoreactivity.
  • Mechanisms of antigenicity loss, such as oxidation and drying, are not fully understood.
  • Optimal conservation methods for extended tissue section storage require further investigation.

Purpose of the Study:

  • To investigate the impact of oxidation and drying on antigenicity in tissue sections.
  • To determine the effects of photo-oxidation and artificial drying on immunohistochemistry (IHC) outcomes.
  • To establish protocols for assessing antigen stability in stored tissue samples.

Main Methods:

  • Paraffin-embedded breast carcinoma sections were exposed to hydrogen peroxide (H2O2), ultraviolet A (UVA) irradiation, and dry heat (56°C).

Related Experiment Videos

  • Immunohistochemistry for oestrogen receptor (ER) was performed after variable exposure times.
  • Stability of multiple antigens (ER, PR, HER-2neu, etc.) was assessed using tissue microarrays subjected to heat and UVA irradiation.

    • Main Results:

    • Extended drying (≥10 days) and UVA irradiation (≥3 days) led to decreased ER staining intensity.
    • Chemical oxidation with H2O2 did not affect ER staining.
    • Most tested antigens (except HER-2neu, PSA, Fli-1) showed reduced antigenicity after heating and UVA exposure, but not complete loss.

    Conclusions:

    • Photo-oxidation and drying significantly influence immunohistochemistry results.
    • The study provides evidence for antigen degradation due to specific storage conditions.
    • Protocols for evaluating the stability of various antigens under stress conditions were developed.