Open Access Highly Accessed Original article

Tip induced fluorescence quenching for nanometer optical and topographical resolution

Olaf Schulz127, Zhao Zhao34, Alex Ward12, Marcelle Koenig5, Felix Koberling5, Yan Liu34, Jörg Enderlein6, Hao Yan34 and Robert Ros12*

Author Affiliations

1 Department of Physics, Arizona State University, Tempe AZ 85287, USA

2 Center for Biological Physics, Arizona State University, Tempe AZ 85287, USA

3 Biodesign Institute, Arizona State University, Tempe AZ 85287, USA

4 Department of Chemistry and Biochemistry, Arizona State University, Tempe AZ 85287, USA

5 PicoQuant GmbH, Rudower Chaussee 29, Berlin 12489, Germany

6 III. Institute of Physics, Georg August University, Göttingen 37077, Germany

7 Present address: III. Institute of Physics, Georg August University, Göttingen 37077, Germany

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Optical Nanoscopy 2013, 2:1  doi:10.1186/2192-2853-2-1

Published: 21 February 2013

Abstract

Progress in nanosciences and life sciences is closely related to developments of high resolution imaging techniques. We introduce a technique which produces correlated topography and fluorescence lifetime images with nanometer resolution. Spot sizes below 5 nm are achieved by quenching of the fluorescence with silicon probes of an atomic force microscope which is combined and synchronized with a confocal fluorescence lifetime microscope. Moreover, we demonstrate the ability to locate and resolve the position of two fluorescent molecules separated by 20.7 nm on a DNA origami triangle with 120 nm side length by correlating topography and fluorescence data. With this method, we anticipate applications in nano- and life sciences, such as the determination of the structure of macromolecular assemblies on surfaces, molecular interactions, as well as the structure and function of nanomaterials.

Keywords:
Correlated atomic force and fluorescence microscopy; Optical ultraresolution microscopy; DNA nanostructures; Single molecule detection