Strategies for improving the immunohistochemical staining of various intranuclear prognostic markers in formalin-paraffin sections: androgen receptor, estrogen receptor, progesterone receptor, p53 protein, proliferating cell nuclear antigen, and Ki-67 antigen revealed by antigen retrieval techniques.
Author(s): Taylor CR, Shi SR, Chaiwun B, Young L, Imam SA, Cote RJ
Publication: Hum Pathol, 1994, Vol. 25, Page 263
PubMed ID: 7512074 PubMed Review Paper? No
Purpose of Paper
Conclusion of Paper
Studies
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Study Purpose
The purpose of this study was to evaluate the effectiveness of different antigen retrieval solutions on restoring antigenicity in FFPE sections of estrogen receptor, progesterone receptor, androgen receptor, p53 protein, proliferating cell nuclear antigen, and Ki-67 antigen, all of which are intranuclear antigens. These were examined in 40 different malignant neoplasms, including breast, colon, stomach, thyroid, lung, and prostate carcinoma, as well as lymphoma, melanoma, seminoma, and some sarcomas, as well as non-neoplastic tonsil.
Summary of Findings:
Antigen retrieval was necessary for all antigens evaluated. Optimal antigen retrieval methods were antigen specific.The authors report that sections pretreated with (a) lead thiocyanate displayed nuclear staining of variable intensity; (b) sodium citrate buffer resulted in the least variable nuclear staining with greatest contrast to background; (c) urea yielded acceptable results with all antigens but considerable background was present in some tissues; (d) distilled water resulted in moderate to poor staining, (e) glycine HCL buffer (without detergent) resulted in non variable nuclear staining with high contrast to background, although the addition of detergent compromised morphology. No differences were observed between antibody visualization with the supersensitive biotin streptavidin or avidin-biotin conjugate kits. Microwave heating for 10 minutes was sufficient in all cases. Optimal results were obtained with tissue fixed for 12 - 24 hours.
Biospecimens
- Tissue - Breast
- Tissue - Stomach
- Tissue - Thyroid Gland
- Tissue - Lung
- Tissue - Colorectal
- Tissue - Prostate
- Tissue - Lymph Node
- Tissue - Skin
- Tissue - Testis
- Tissue - Tonsil
- Tissue - Other
Preservative Types
- Formalin
Diagnoses:
- Neoplastic - Carcinoma
- Neoplastic - Lymphoma
- Not specified
- Neoplastic - Melanoma
- Neoplastic - Other
- Neoplastic - Sarcoma
Platform:
Analyte Technology Platform Protein Immunohistochemistry Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Analyte Extraction and Purification Antigen retrieval Glycine HCl buffer
Lead thiocyanate
Sodium citrate buffer
Urea
Distilled water
Glycine HCl buffer with detergent
Biospecimen Preservation Time in fixative 12 h
24 h
3 d
7 d
Analyte Extraction and Purification Temperature of heat-induced retrieval 3 min of microwave heating
5 min of microwave heating
10 min of microwave heating
20 min of microwave heating
Immunohistochemistry Specific Detection method Supersensitive biotin streptavidin kit
Avidin-biotin conjugate kit