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A plasmonic ‘antenna-in-box’ platform for enhanced single-molecule analysis at micromolar concentrations


Single-molecule fluorescence techniques1,2,3 are key for a number of applications, including DNA sequencing4,5, molecular and cell biology6,7 and early diagnosis8. Unfortunately, observation of single molecules by diffraction-limited optics is restricted to detection volumes in the femtolitre range and requires pico- or nanomolar concentrations, far below the micromolar range where most biological reactions occur2. This limitation can be overcome using plasmonic nanostructures, which enable the confinement of light down to nanoscale volumes9,10,11,12,13. Although these nanoantennas enhance fluorescence brightness14,15,16,17,18,19,20, large background signals20,21,22 and/or unspecific binding to the metallic surface23,24,25 have hampered the detection of individual fluorescent molecules in solution at high concentrations. Here we introduce a novel ‘antenna-in-box’ platform that is based on a gap-antenna inside a nanoaperture. This design combines fluorescent signal enhancement and background screening, offering high single-molecule sensitivity (fluorescence enhancement up to 1,100-fold and microsecond transit times) at micromolar sample concentrations and zeptolitre-range detection volumes. The antenna-in-box device can be optimized for single-molecule fluorescence studies at physiologically relevant concentrations, as we demonstrate using various biomolecules.

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Figure 1: Antenna-in-box platform for single-molecule analysis at high concentrations.
Figure 2: Enhanced single-molecule analysis with an antenna-in-box.
Figure 3: Fluorescence enhancement and volume reduction as a function of nanoantenna gap size.
Figure 4: Applicability of the antenna-in-box to detect and discriminate individual biomolecules at 10 µM concentrations.


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The research leading to these results received funding from the European Commission's Seventh Framework Programme (FP7-ICT-2011-7) under grant agreements 288263 (NanoVista), ERC StG 278242 (ExtendFRET) and ERC AdG (NanoAntennas), the Spanish Ministry of Science and Innovation and the Agence Nationale de la Recherche under grant ANR-10-INBS-04-01 (France Bio Imaging). The authors thank A. Brisson for providing the Annexin sample.

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J.W., H.R., N.F.v.H. and M.G-P. conceived and designed the experiments. D.P., J.W. and S.B.M. performed the experiments and analysed the data. M.M. and T.S.v.Z. fabricated the antennas. J.W., M.G-P. and N.F.v.H. wrote the manuscript.

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Correspondence to Jérôme Wenger.

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The authors declare no competing financial interests.

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Punj, D., Mivelle, M., Moparthi, S. et al. A plasmonic ‘antenna-in-box’ platform for enhanced single-molecule analysis at micromolar concentrations. Nature Nanotech 8, 512–516 (2013).

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