Polarized gonio fluorimetric measurements of DNA-chip: a step forward towards fluorescence quantification

Pierre Barritault; Stephane Getin; Patrick Chaton; Francoise Vinet; Brigitte Fouque; Jean-Louis P. Stehle

doi:10.1117/12.472090

21 June 2002 Polarized gonio fluorimetric measurements of DNA-chip: a step forward towards fluorescence quantification

Pierre Barritault, Stephane Getin, Patrick Chaton, Francoise Vinet, Brigitte Fouque, Jean-Louis P. Stehle

Proceedings Volume 4626, Biomedical Nanotechnology Architectures and Applications; (2002) https://doi.org/10.1117/12.472090
Event: International Symposium on Biomedical Optics, 2002, San Jose, CA, United States

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.

Citation Download Citation

Pierre Barritault, Stephane Getin, Patrick Chaton, Francoise Vinet, Brigitte Fouque, and Jean-Louis P. Stehle "Polarized gonio fluorimetric measurements of DNA-chip: a step forward towards fluorescence quantification", Proc. SPIE 4626, Biomedical Nanotechnology Architectures and Applications, (21 June 2002); https://doi.org/10.1117/12.472090

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