title: Determination of surface-bound-fluorophore orientation by goniometric fluorescence polarization: application to quantification of DNA-chip readouts.
abstract: We present a polarized goniofluorimeter designed to measure the observation-angle and polarization-dependent intensity emitted by a group of surface-bound fluorescent molecules. We studied two types of surface bonding: In one case, dyes were adsorbed into the surface by spin coating, and in the other, dyes were covalently immobilized to DNA strands. Fluorescent dyes consisted of Cy3 and Alexa546. The substrate was a silicon wafer bearing a silicon dioxide layer. The different samples presented a wide panel of reproducible experimental behavior. By confronting experimental behavior with theory and simulation, we can explain these differences as directly linked to the mean orientation of fluorophores with respect to the surface.
authors.affiliation: Département d'Optronique, Commissariat à l'Energie Atomique, Grenoble, France. barritault@chartreuse.cea.fr
authors.forename: Pierre
authors.initials: P
authors.lastname: Barritault
authors.forename: Stéphane
authors.initials: S
authors.lastname: Gétin
authors.forename: Patrick
authors.initials: P
authors.lastname: Chaton
authors.forename: Françoise
authors.initials: F
authors.lastname: Vinet
authors.forename: Brigitte
authors.initials: B
authors.lastname: Fouqué
mesh_terms: D003198:Computer Simulation; D005454:Fluorescence Polarization; D005456:Fluorescent Dyes; D006801:Humans; D008962:Models, Theoretical; D020411:Oligonucleotide Array Sequence Analysis
chemical_list: D005456:Fluorescent Dyes