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Active substrates improving sensitivity in biomedical fluorescence microscopy

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 setup. 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 setup 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.
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Submitted on : Tuesday, March 24, 2020 - 10:41:49 PM
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Eric Le Moal, Sandrine Lévêque-Fort, Emmanuel Fort, J.-P. Lacharme, M.-P. Fontaine-Aupart, et al.. Active substrates improving sensitivity in biomedical fluorescence microscopy. European Conference on Biomedical Optics 2005, Jun 2005, Munich, Germany. pp.58600G, ⟨10.1117/12.632945⟩. ⟨hal-02518053⟩

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