Automated immunohistochemical staining of formalin-fixed and paraffin-embedded tissues using a catalyzed signal amplification method.
An immunohistochemical assay using catalyzed signal amplification (CSA), which is based on the peroxidase catalyzed deposition of biotinylated tyramide, is a highly sensitive method to visualize weak immunohistochemical signals originating from rare antigens or masked antigens in formalin-fixed, paraffin-embedded (FFPE) tissues. However, CSA methods are hampered by poor reproducibility and the complexity of their staining procedures. In this study, we aimed to apply the CSA procedure to a capillary gap-based, automated immunostainer, TechMate Horizon, to perform immunohistochemical signal amplification effectively and reproducibly. A variety of cellular antigens previously considered to be undetectable in FFPE human specimens were selected and examined with the automated immunostainer. Compared with the manual CSA staining method that takes more than 2 hours, the automated CSA method took less than 2 hours to complete. The staining results from the automated CSA method presented higher reproducibility, as well as lower background owing to well-regulated, punctual staining and washing at every step of the procedure. Conclusively, the automation of the CSA method enabled us to perform the time-consuming and complicated CSA amplification technique with minimal effort in an accurate, consistent, and reproducible manner.
Department of Immunohistochemistry, DAKO Japan, Kyoto.
K
K
Hashizume
Y
Y
Hatanaka
Y
Y
Kamihara
Y
Y
Tani
D000941:Antigens / Q000378:metabolism; D001943:Breast Neoplasms / Q000276:immunology / Q000473:pathology; D003110:Colonic Neoplasms / Q000276:immunology / Q000473:pathology; D005404:Fixatives; D005557:Formaldehyde; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D014066:Palatine Tonsil / Q000033:anatomy & histology / Q000276:immunology; D016612:Paraffin Embedding; D015203:Reproducibility of Results
D000941:Antigens; D005404:Fixatives; D005557:Formaldehyde