25 February 2010 Fluorescence correlation spectroscopy on nano-fakir surfaces
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Abstract
Single biomolecule behaviour can reveal crucial information about processes not accessible by ensemble measurements. It thus represents a real biotechnological challenge. Common optical microscopy approaches require pico- to nano-molar concentrations in order to isolate an individual molecule in the observation volume. However, biologically relevant conditions often involve micromolar concentrations, which impose a drastic reduction of the conventional observation volume by at least three orders of magnitude. This confinement is also crucial for mapping sub-wavelength heterogeneities in cells, which play an important role in many biological processes. We propose an original approach, which couples Fluorescence Correlation Spectroscopy (FCS), a powerful tool to retrieve essential information on single molecular behaviour, and nano-fakir substrates with strong field enhancements and confinements at their surface. These electromagnetic singularities at nanometer scale, called "hotspots", are the result of the unique optical properties of surface plasmons. They provide an elegant means for studying single-molecule dynamics at high concentrations by reducing dramatically the excitation volume and enhancing the fluorophore signal by several orders of magnitude. The nano-fakir substrates used are obtained from etching optical fiber bundles followed by sputtering of a gold thin-film. It allows one to design reproducible arrays of nanotips.
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Julie Delahaye, Julie Delahaye, Samuel Gresillon, Samuel Gresillon, Sandrine Lévêque-Fort, Sandrine Lévêque-Fort, Neso Sojic, Neso Sojic, Emmanuel Fort, Emmanuel Fort, } "Fluorescence correlation spectroscopy on nano-fakir surfaces", Proc. SPIE 7571, Single Molecule Spectroscopy and Imaging III, 757114 (25 February 2010); doi: 10.1117/12.842763; https://doi.org/10.1117/12.842763
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