title: Antigen retrieval, blocking, detection and visualisation systems in immunohistochemistry: a review and practical evaluation of tyramide and rolling circle amplification systems.
abstract: To achieve specificity and sensitivity using immunohistochemistry it is necessary to combine the application of validated primary antibodies with optimised pre-treatment, detection and visualisation steps. The influence of these surrounding procedures is reviewed. A practical evaluation of tyramide signal amplification and rolling circle amplification detection methods is provided in which formalin fixed paraffin embedded sections of adenocarcinomas of breast, colon and lung together with squamous metaplasia of lung were immunostained with CD20 and CK19 primary antibodies. The results indicate that the detection systems are of comparable sensitivity and specificity.
authors.affiliation: University of Westminster, 115 New Cavendish Street, London W1W 6UW, United Kingdom. Electronic address: a.warford@westminster.ac.uk.
authors.forename: Anthony
authors.initials: A
authors.lastname: Warford
authors.affiliation: University of Westminster, 115 New Cavendish Street, London W1W 6UW, United Kingdom.
authors.forename: Hameed
authors.initials: H
authors.lastname: Akbar
authors.affiliation: University of Westminster, 115 New Cavendish Street, London W1W 6UW, United Kingdom.
authors.forename: Deise
authors.initials: D
authors.lastname: Riberio
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology; D000941:Antigens / Q000737:chemistry*; D018951:Antigens, CD20 / Q000737:chemistry; D001943:Breast Neoplasms / Q000276:immunology; D003110:Colonic Neoplasms / Q000276:immunology; D005260:Female; D005557:Formaldehyde / Q000737:chemistry; D006358:Hot Temperature; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D008175:Lung Neoplasms / Q000276:immunology; D016612:Paraffin Embedding; D013696:Temperature; D016707:Tissue Fixation; D014439:Tyramine / Q000737:chemistry*
chemical_list: D000906:Antibodies; D000941:Antigens; D018951:Antigens, CD20; D005557:Formaldehyde; D014439:Tyramine
keywords: Antigen retrieval; Detection systems; Immunohistochemistry; Rolling circle amplification; Tyramide signal amplification