Optical microscopy using a single-molecule light source

Abstract

Rapid progress in science on nanoscopic scales has promoted increasing interest in techniques of ultrahigh-resolution optical microscopy. The diffraction limit can be surpassed by illuminating an object in the near field through a sub-wavelength aperture at the end of a sharp metallic probe1,2. Proposed modifications3,4 of this technique involve replacing the physical aperture by a nanoscopic active light source. Advances in the spatial5 and spectral6 detection of individual fluorescent molecules, using near-field and far-field methods7, suggest the possibility of using a single molecule8,9 as the illumination source. Here we present optical images taken with a single molecule as a point-like source of illumination, by combining fluorescence excitation spectroscopy10 with shear-force microscopy11. Our single-molecule probe has potential for achieving molecular resolution in optical microscopy; it should also facilitate controlled studies of nanometre-scale phenomena (such as resonant energy transfer) with improved lateral and axial spatial resolution.

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Figure 1: Overview of experimental features for realizing a single-molecule probe.
Figure 2: Topography and optical images of the sample.
Figure 3: Images taken with the same molecule positioned at four different distances from the sample.

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Acknowledgements

We are grateful to O. Martin and C. Henkel for many fruitful discussions. We thank B. Eiermann for contribution to the initial phase of the experiment, T. Kalkbrenner for the fabrication of the sample, and H. M. Ludwig for the AFM image in Fig. 2. We also thank P. Leiderer and his co-workers, and colleagues at Omicron Vakuum GmbH for help and advice regarding the construction of the piezo-driven translation stages. J. M. acknowledges a fellowship from the Carl-Zeiss-Schott Förderstiftung. This work was supported by the Deutsche Forschungsgemeinschaft and the Bundesministerium für Bildung und Forschung.

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Correspondence to V. Sandoghdar.

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Michaelis, J., Hettich, C., Mlynek, J. et al. Optical microscopy using a single-molecule light source. Nature 405, 325–328 (2000). https://doi.org/10.1038/35012545

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