title: Interfacial polymerization for colorimetric labeling of protein expression in cells.
abstract: Determining the location of rare proteins in cells typically requires the use of on-sample amplification. Antibody based recognition and enzymatic amplification is used to produce large amounts of visible label at the site of protein expression, but these techniques suffer from the presence of nonspecific reactivity in the biological sample and from poor spatial control over the label. Polymerization based amplification is a recently developed alternative means of creating an on-sample amplification for fluorescence applications, while not suffering from endogenous labels or loss of signal localization. This manuscript builds upon polymerization based amplification by developing a stable, archivable, and colorimetric mode of amplification termed Polymer Dye Labeling. The basic concept involves an interfacial polymer grown at the site of protein expression and subsequent staining of this polymer with an appropriate dye. The dyes Evans Blue and eosin were initially investigated for colorimetric response in a microarray setting, where both specifically stained polymer films on glass. The process was translated to the staining of protein expression in human dermal fibroblast cells, and Polymer Dye Labeling was specific to regions consistent with desired protein expression. The labeling is stable for over 200 days in ambient conditions and is also compatible with modern mounting medium.
authors.affiliation: Department of Chemical and Materials Engineering, University of Kentucky, Lexington, Kentucky, United States of America.
authors.forename: Jacob L
authors.initials: JL
authors.lastname: Lilly
authors.affiliation: Department of Chemical and Materials Engineering, University of Kentucky, Lexington, Kentucky, United States of America.
authors.forename: Phillip R
authors.initials: PR
authors.lastname: Sheldon
authors.affiliation: Department of Pediatrics, University of Colorado, Denver, Colorado, United States of America.
authors.forename: Liv J
authors.initials: LJ
authors.lastname: Hoversten
authors.affiliation: Department of Chemical and Materials Engineering, University of Kentucky, Lexington, Kentucky, United States of America.
authors.forename: Gabriela
authors.initials: G
authors.lastname: Romero
authors.affiliation: Department of Pediatrics, University of Colorado, Denver, Colorado, United States of America.
authors.forename: Vivek
authors.initials: V
authors.lastname: Balasubramaniam
authors.affiliation: Department of Chemical and Materials Engineering, University of Kentucky, Lexington, Kentucky, United States of America.
authors.forename: Brad J
authors.initials: BJ
authors.lastname: Berron
mesh_terms: D003124:Colorimetry / Q000379:methods; D004396:Coloring Agents / Q000737:chemistry*; D004801:Eosine Yellowish-(YS) / Q000737:chemistry*; D005070:Evans Blue / Q000737:chemistry*; D005347:Fibroblasts / Q000166:cytology*; D006801:Humans; D058105:Polymerization; D011506:Proteins / Q000032:analysis*; D013194:Staining and Labeling
chemical_list: D004396:Coloring Agents; D011506:Proteins; D005070:Evans Blue; D004801:Eosine Yellowish-(YS)