title: High-resolution immunophenotyping of subcellular compartments in tissue microarrays by enzyme metallography.
abstract: Ultrasensitive bright field in situ hybridization assays using enzyme metallography (EnzMet) have been developed and validated, but little is known regarding the applicability of EnzMet for immunophenotypic detection of protein via IHC. Superior resolution via discrete metallographic deposits offers the potential for enhancing high-resolution immunophenotyping. Using high-complexity tissue microarrays (TMAs), 88 common solid tumors were evaluated by automated EnzMet (Nanoprobes and Ventana). Targets were chosen to assess the ability of EnzMet to specifically localize encoded antigens in the nucleus (estrogen receptor), cytoplasm (cytokeratins), and cytoplasmic membrane (HER2) in TMAs. Results were compared with conventional IHC diaminobenzidine (DAB) immunostaining. There was full concordance between the EnzMet and conventional IHC results. Furthermore, the EnzMet reaction products did not appreciably diffuse, were dense and sharply defined, and provided excellent high-resolution differentiation of cellular compartments in paraffin sections for the nuclear, cytoplasmic, and cell membrane-localized antigens evaluated. The higher density of elemental silver deposited during enzyme metallography permitted evaluation of core immunophenotypes at a relatively low magnification, allowing more tissue to be screened in an efficient manner. This preliminary study shows the utility of using enzyme metallography for high-resolution immunophenotyping in TMAs.
authors.affiliation: Department of Anatomic and Clinical Pathology, the Cleveland Clinic Foundation, OH 44195, USA. TubbsR@ccf.org
authors.forename: Raymond
authors.initials: R
authors.lastname: Tubbs
authors.forename: James
authors.initials: J
authors.lastname: Pettay
authors.forename: Richard
authors.initials: R
authors.lastname: Powell
authors.forename: David G
authors.initials: DG
authors.lastname: Hicks
authors.forename: Patrick
authors.initials: P
authors.lastname: Roche
authors.forename: William
authors.initials: W
authors.lastname: Powell
authors.forename: Thomas
authors.initials: T
authors.lastname: Grogan
authors.forename: James F
authors.initials: JF
authors.lastname: Hainfeld
mesh_terms: D001943:Breast Neoplasms / Q000276:immunology / Q000473:pathology; D002277:Carcinoma / Q000276:immunology / Q000473:pathology*; D002451:Cell Compartmentation / Q000276:immunology; D022082:Cellular Structures / Q000276:immunology / Q000473:pathology*; D003110:Colonic Neoplasms / Q000276:immunology / Q000473:pathology; D005260:Female; D006801:Humans; D016130:Immunophenotyping / Q000379:methods*; D007680:Kidney Neoplasms / Q000276:immunology / Q000473:pathology; D008297:Male; D015335:Molecular Probes / Q000276:immunology; D009369:Neoplasms / Q000276:immunology / Q000473:pathology*; D011471:Prostatic Neoplasms / Q000276:immunology / Q000473:pathology; D012189:Retrospective Studies; D012834:Silver / Q000276:immunology; D046888:Tissue Array Analysis
chemical_list: D015335:Molecular Probes; D012834:Silver