Antigenicity testing by immunohistochemistry after tissue oxidation.
Author(s): Blind C, Koepenik A, Pacyna-Gengelbach M, Fernahl G, Deutschmann N, Dietel M, Krenn V, Petersen I
Publication: J Clin Pathol, 2008, Vol. 61, Page 79-83
PubMed ID: 17412873 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 the effects of storing cut sections prior to coating slides with paraffin on IHC staining for ER and HER2 in breast carcinoma specimens. Slides were stored at room temperature for various durations and then dipped into liquid paraffin and stored for an additional 3 months. They were then incubated at 58 degrees C overnight before the paraffin coating was removed.
Summary of Findings:
For all storage durations, the authors state that coating the slides with paraffin wax made deparaffinization difficult and gave irregular IHC results due to the retention of a thin paraffin layer on the slides, and they recommend against this paraffin coating technique.
Biospecimens
Preservative Types
- Formalin
Diagnoses:
- Neoplastic - Carcinoma
Platform:
Analyte Technology Platform Protein Immunohistochemistry Protein Tissue microarray Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Storage Time at room temperature 0 d
1 d
2 d
4 d
7 d
10 d
14 d
21 d
30 d
45 d
60 d
3 months
Storage Storage conditions Slide coated with paraffin wax
Slide not coated with paraffin wax
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Study Purpose
The purpose of this study was to determine the effects oxidation induced by H2O2, storage at 56 degrees C, or UV irradiation on IHC staining for 15 antigens in breast carcinoma specimens. Slide-mounted FFPE sections were stored in a H2O2 solution for 1, 4, or 24 h; stored at 56°C (dry heat) for 1 day to 12 weeks; or under a UVA lamp for 30 min to 8 weeks prior to analysis. Positive control specimens used for some of the tested antigens included myometrium, ovary, larynx, uterus, stomach, placenta, lymph node, bone, prostate, pancreas, liver, and skin.
Summary of Findings:
The only antigens unaffected by either dry heat for 4 weeks or UV treatment for 7 d were HER2, Fli-1, and PSA. CK5/6, CK7, and CK20 IHC staining was decreased in response to the UV irradiation but not the dry heat. IHC staining of the remaining 9 antigens (ER, progesterone receptor (PR), p53, p63, p16, smooth muscle antigen (SMA), c-kit, CD20, and epidermal growth factor receptor (EGFR)) was decreased by both of these treatments. ER immunostaining was reduced in slides subjected to dry heat (56 degrees C) for 10 days up to 12 weeks when compared to slides stored for the same duration at room temperature and after 3-4 days of subjecting the slides to UV treatment. Although ER antigenicity was not entirely lost, tissue destruction was apparent after 6 to 8 weeks of UV exposure. None of the tested concentrations or durations of H2O2 treatment had an effect on ER staining intensity, however, after 24 hours of H2O2 treatment, partial corrosion of the specimens was apparent and indirectly affected the ability of researchers to assess IHC results.
Biospecimens
- Tissue - Breast
- Tissue - Uterus
- Tissue - Larynx
- Tissue - Stomach
- Tissue - Skin
- Tissue - Lymph Node
- Tissue - Bone
- Tissue - Prostate
- Tissue - Pancreas
- Tissue - Liver
- Tissue - Ovary
- Tissue - Placenta
Preservative Types
- Formalin
Diagnoses:
- Neoplastic - Carcinoma
- Neoplastic - Benign
- Neoplastic - Lymphoma
- Neoplastic - Sarcoma
- Fibroma/Fibroid
- Normal
Platform:
Analyte Technology Platform Protein Immunohistochemistry Protein Tissue microarray Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Storage Storage conditions 0.3% H2O2
3% H2O2
30% H2O2
No H2O2 treatment
Under UV light
Protected from daylight
Storage Storage duration 1 h
4 h
24 h
1 d-12 weeks
30 min-8 weeks
Storage Storage temperature Room temperature
56 degrees C
Immunohistochemistry Specific Targeted peptide/protein HER2
Fli-1
PSA
CK5/6
CK7
CK20
ER
PR
p53
p63
p16
SMA
c-kit
CD20
EGFR