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Proceedings Paper

Polarized gonio fluorimetric measurements of DNA-chip: a step forward towards fluorescence quantification
Author(s): Pierre Barritault; Stephane Getin; Patrick Chaton; Francoise Vinet; Brigitte Fouque; Jean-Louis P. Stehle
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Paper Abstract

Quantifying hybridization and therefore fluorescence signals has become a key-issue in DNA-chip technology. Thus a better understanding of fluorescence near a surface has become a necessity. To study this issue, we modeled the fluorophore after an electromagnetic dipole radiating over the substrate; we then developed a simulation code which enabled us to calculate the observation-angle-dependent-intensity radiated by a population and altitude. In the mean time we developed a polarized-gonio-fluoimeter which permits angular fluorescence patterns and fluorescence polarization measurements. We studied DNA-chips obtained by covalent grafting of labeled oligonucleotides. Simulation curves perfectly matched experimental ones, enabling an accurate determination of fluorophore localization on the substrate. Once achieved a better understanding of the fluorophore emission, we designed and realized a thin-film-coated microscope slide dedicated to the enhancement of DNA-chip fluorescence. This substrate was used in a c-DNA gene analysis. Fluorescence enhancement was clearly observed enabling the detection of Cart1 and P2A which are undetectable when using non-coated microscope slides.

Paper Details

Date Published: 21 June 2002
PDF: 8 pages
Proc. SPIE 4626, Biomedical Nanotechnology Architectures and Applications, (21 June 2002); doi: 10.1117/12.472090
Show Author Affiliations
Pierre Barritault, CEA-LETI (France)
Stephane Getin, CEA-LETI (France)
Patrick Chaton, CEA-LETI (France)
Francoise Vinet, CEA-LETI (France)
Brigitte Fouque, CEA-DSV (France)
Jean-Louis P. Stehle, SOPRA SA (France)

Published in SPIE Proceedings Vol. 4626:
Biomedical Nanotechnology Architectures and Applications
Raymond P. Mariella; Catherine J. Murphy; Michelle Palmer; David A. Dunn; Darryl J. Bornhop; David A. Dunn; Raymond P. Mariella; Catherine J. Murphy; Dan V. Nicolau; Shuming Nie; Michelle Palmer; Ramesh Raghavachari; Darryl J. Bornhop; Ramesh Raghavachari; Shuming Nie; Ramesh Raghavachari, Editor(s)

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