title: Covalently deposited dyes: a new chromogen paradigm that facilitates analysis of multiple biomarkers in situ.
abstract: Multiplexed analysis of multiple biomarkers in a tissue sample requires use of reporter dyes with specific spectral properties that enable discrimination of signals. Conventional chromogens with broad absorbance spectra, widely used in immunohistochemistry (IHC), offer limited utility for multiplexed detection. Many dyes with narrow absorbance spectra, eg rhodamines, fluoresceins, and cyanines, potentially useful for multiplexed detection are well-characterized; however, generation of a chromogenic reagent useful for IHC analysis has not been demonstrated. Studies reported herein demonstrate utility of tyramine-chemistry for synthesis of a wide variety of new chromogenic dye conjugates useful for multiplexed in situ analysis using conventional light microscopes. The dyes, useful individually or in blends to generate new colors, provide signal sensitivity and dynamic range similar to conventional DAB chromogen, while enabling analysis of co-localized biomarkers. It is anticipated that this new paradigm will enable generation of a wide variety of new chromogens, useful for both research and clinical biomarker analysis that will benefit clinicians and patients.
authors.affiliation: Tissue Research and Early Development, Ventana Medical Systems, Inc., Tucson, AZ, USA.
authors.forename: William A
authors.initials: WA
authors.lastname: Day
authors.affiliation: Tissue Research and Early Development, Ventana Medical Systems, Inc., Tucson, AZ, USA.
authors.forename: Mark R
authors.initials: MR
authors.lastname: Lefever
authors.affiliation: Tissue Research and Early Development, Ventana Medical Systems, Inc., Tucson, AZ, USA.
authors.forename: Robert L
authors.initials: RL
authors.lastname: Ochs
authors.affiliation: Tissue Research and Early Development, Ventana Medical Systems, Inc., Tucson, AZ, USA.
authors.forename: Anne
authors.initials: A
authors.lastname: Pedata
authors.affiliation: Tissue Research and Early Development, Ventana Medical Systems, Inc., Tucson, AZ, USA.
authors.forename: Lauren J
authors.initials: LJ
authors.lastname: Behman
authors.affiliation: Tissue Research and Early Development, Ventana Medical Systems, Inc., Tucson, AZ, USA.
authors.forename: Julia
authors.initials: J
authors.lastname: Ashworth-Sharpe
authors.affiliation: Tissue Research and Early Development, Ventana Medical Systems, Inc., Tucson, AZ, USA.
authors.forename: Donald D
authors.initials: DD
authors.lastname: Johnson
authors.affiliation: Tissue Research and Early Development, Ventana Medical Systems, Inc., Tucson, AZ, USA.
authors.forename: Eric J
authors.initials: EJ
authors.lastname: May
authors.affiliation: Tissue Research and Early Development, Ventana Medical Systems, Inc., Tucson, AZ, USA.
authors.forename: James G
authors.initials: JG
authors.lastname: Grille
authors.affiliation: Tissue Research and Early Development, Ventana Medical Systems, Inc., Tucson, AZ, USA.
authors.forename: Esteban A
authors.initials: EA
authors.lastname: Roberts
authors.affiliation: Tissue Research and Early Development, Ventana Medical Systems, Inc., Tucson, AZ, USA.
authors.forename: Jerry W
authors.initials: JW
authors.lastname: Kosmeder
authors.affiliation: Tissue Research and Early Development, Ventana Medical Systems, Inc., Tucson, AZ, USA.
authors.forename: Larry E
authors.identifier: 0000-0002-5813-0110
authors.initials: LE
authors.lastname: Morrison
mesh_terms: D015100:3,3'-Diaminobenzidine / Q000737:chemistry; D015415:Biomarkers / Q000032:analysis* / Q000737:chemistry; D002863:Chromogenic Compounds / Q000138:chemical synthesis / Q000737:chemistry*; D004396:Coloring Agents / Q000138:chemical synthesis / Q000737:chemistry*; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D017403:In Situ Hybridization / Q000379:methods*; D008956:Models, Chemical; D015394:Molecular Structure; D015203:Reproducibility of Results; D014439:Tyramine / Q000737:chemistry
chemical_list: D015415:Biomarkers; D002863:Chromogenic Compounds; D004396:Coloring Agents; D015100:3,3'-Diaminobenzidine; D014439:Tyramine