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Proceedings Paper

Active substrates improving sensitivity in biomedical fluorescence microscopy
Author(s): E. Le Moal; S. Leveque-Fort; E. Fort; J.-P. Lacharme; M.-P. Fontaine-Aupart; C. Ricolleau
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Paper Abstract

Fluorescence is widely used as a spectroscopic tool or for biomedical imaging, in particular for DNA chips. In some cases, detection of very low molecular concentrations and precise localization of biomarkers are limited by the weakness of the fluorescence signal. We present a new method based on sample substrates that improve fluorescence detection sensitivity. These active substrates consist in glass slides covered with metal (gold or silver) and dielectric (alumina) films and can directly be used with common microscope set-up. Fluorescence enhancement affects both excitation and decay rates and is strongly dependant on the distance to the metal surface. Furthermore, fluorescence collection is improved since fluorophore emission lobes are advantageously modified close to a reflective surface. Finally, additional improvements are achieved by structuring the metallic layer. Substrates morphology was mapped by Atomic Force Microscopy (AFM). Substrates optical properties were studied using mono- and bi-photonic fluorescence microscopy with time resolution. An original set-up was implemented for spatial radiation pattern's measurement. Detection improvement was then tested on commercial devices. Several biomedical applications are presented. Enhancement by two orders of magnitude are achieved for DNA chips and signal-to-noise ratio is greatly increased for cells imaging.

Paper Details

Date Published: 8 October 2005
PDF: 10 pages
Proc. SPIE 5860, Confocal, Multiphoton, and Nonlinear Microscopic Imaging II, 58600G (8 October 2005); doi: 10.1117/12.632945
Show Author Affiliations
E. Le Moal, Lab. Materiaux et Phenomenes Quantiques, Univ. Paris 7, Lab. Physique du Solide, ESPCI (France)
Lab. PhotoPhysique Moleculaire, CNRS (France)
S. Leveque-Fort, Lab. PhotoPhysique Moleculaire, CNRS (France)
E. Fort, Lab. Materiaux et Phenomenes Quantiques, Univ. Paris 7, Lab. Physique du Solide, ESPCI (France)
J.-P. Lacharme, Lab. Materiaux et Phenomenes Quantiques, Univ. Paris 7, Lab. Physique du Solide, ESPCI (France)
M.-P. Fontaine-Aupart, Lab. PhotoPhysique Moleculaire, CNRS (France)
C. Ricolleau, Lab. PhotoPhysique Moleculaire, CNRS (France)


Published in SPIE Proceedings Vol. 5860:
Confocal, Multiphoton, and Nonlinear Microscopic Imaging II
Tony Wilson, Editor(s)

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