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Tuesday, 24 April, 2012

Distance Dependence of Single-Fluorophore Quenching by Gold Nanoparticles Studied on DNA Origami

G. P. Acuna, M. Bucher, I. H. Stein, C. Steinhauer, A. Kuzyk, P. Holzmeister, R. Schreiber, A. Moroz, F. D. Stefani, T. Liedl, F. C. Simmel, and P. Tinnefeld -
ACS Nano, Vol. 6 (4), pp. 3189-3195 (2012)

We study the distance-dependent quenching of fluorescence due to a metallic nanoparticle in proximity of a fluorophore. In our single-molecule measurements, we achieve excellent control over structure and stoichiometry by using self-assembled DNA structures (DNA origami) as a breadboard where both the fluorophore and the 10 nm metallic nanoparticle are positioned with nanometer precision. The single-molecule spectroscopy method employed here reports on the co-localization of particle and dye, while fluorescence lifetime imaging is used to directly obtain the correlation of intensity and fluorescence lifetime for varying particle to dye distances. Our data can be well explained by exact calculations that include dipole–dipole orientation and distances. Fitting with a more practical model for nanosurface energy transfer yields 10.4 nm as the characteristic distance of 50% energy transfer. The use of DNA nanotechnology together with minimal sample usage by attaching the particles to the DNA origami directly on the microscope coverslip paves the way for more complex experiments exploiting dye–nanoparticle interactions.

 

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