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Article Dans Une Revue ACS Applied Materials & Interfaces Année : 2017

DNA-Guided Delivery of Single Molecules into Zero-Mode Waveguides

Résumé

Zero-mode waveguides (ZMWs) are powerful analytical tools corresponding to optical nanostructures fabricated in a thin metallic film capable of confining an excitation volume to the range of attoliters. This small volume of confinement allows single-molecule fluorescence experiments to be performed at physiologically relevant concentrations of fluorescently labelled biomolecules. Exactly one molecule to be studied must be attached at the floor of the ZMW for signal detection and analysis; however, the massive parallelism of these nanoarrays suffers from a Poissonian-limited distribution of these biomolecules. To date, there is no method available that provides full single molecule occupancy of massively arrayed ZMWs. Here we report the performance of a DNA-guided method that uses steric exclusion properties of large DNA molecules to bias the Poissonian-limited delivery of single molecules. Non-Poissonian statistics were obtained with DNA molecules that contain a free-biotinylated extremity for efficient binding to the floor of the ZMW, which resulted in a decrease of accessibility for a second molecule. Both random coil and condensed DNA conformations drove non-Poissonian single-molecule delivery into ZMWs arrays. The results suggest that an optimal balance between rigidity and flexibility of the macromolecule is critical for favourable accessibility and single occupancy. The optimized method provides means for full exploitation of these massively parallelized analytical tools.

Dates et versions

hal-02391699 , version 1 (03-12-2019)

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Thomas Plénat, Satoko Yoshizawa, Dominique Fourmy. DNA-Guided Delivery of Single Molecules into Zero-Mode Waveguides. ACS Applied Materials & Interfaces, 2017, ⟨10.1021/acsami.7b11953⟩. ⟨hal-02391699⟩
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