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Optically monitoring the mechanical assembly of single molecules

Nature Nanotechnology volume 4, pages 45–49 (2009)Cite this article

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Abstract

Bottom-up assembly at the level of individual molecules requires a combination of utmost spatial precision and efficient monitoring. We have previously shown how to ‘cut-and-paste’ single molecules1, and other groups have demonstrated that it is possible to beat the diffraction limit in optical microscopy2,3,4. Here we show that a combination of single-molecule cut-and-paste surface assembly1, total internal reflection fluorescence microscopy and atomic force microscopy5,6,7,8 can be used to deposit individual fluorophores in well-defined nanoscale patterns and also to monitor the process in real time with nanometre precision. Although the size of the pattern is well below the optical resolution of the microscope, the individual dyes are identified by localizing the centroids and detecting the photobleaching of the fluorophores. With this combination of methods, individual dyes or labelled biomolecules can be arranged at will for specific functions, such as coupled fluorophore systems or tailored enzyme cascades, and monitored with nanoscale precision.

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Figure 1: Monitoring the deposition of a single molecule by AFM and TIRF microscopy.
Figure 2: Localizing single molecules with nanometre precision.
Figure 3: Positional accuracy of the SMCP process.
Figure 4: Determining the position of deposited dyes by photobleaching.

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  • 08 December 2008

    In the version of this Letter originally published online, reference 27 was missing its full details. This has now been corrected in the HTML and PDF versions.

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Acknowledgements

We thank P. Tinnefeld and A. Fornof for helpful discussions. This work was supported by the German Science Foundation, the Nanosystems Initiative Munich (NIM) and Functional Nanosystems (FuNS).

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Authors and Affiliations

  1. Centre for Nanoscience and Physics Department, University of Munich, Amalienstrasse 54, Munich, 80799, Germany

    Stefan K. Kufer, Mathias Strackharn, Stefan W. Stahl, Hermann Gumpp, Elias M. Puchner & Hermann E. Gaub

  2. Centre for Integrated Protein Science Munich, Butenandtstrasse 5 - 13, Munich, 81377, Germany

    Elias M. Puchner

Authors
  1. Stefan K. Kufer
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  2. Mathias Strackharn
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  3. Stefan W. Stahl
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  6. Hermann E. Gaub
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Contributions

S.K.K. and H.E.G. conceived and designed the experiments and co-wrote the paper. S.K.K., M.S. and S.W.S. performed the experiments. S.K.K. and E.M.P. analysed the data. M.S., S.W.S., H.G. and E.M.P. built the AFM–TIRF setup. M.S., S.K.K. and H.E.G. developed the theoretical model of the SMCP spatial uncertainty.

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Correspondence to Hermann E. Gaub.

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Kufer, S., Strackharn, M., Stahl, S. et al. Optically monitoring the mechanical assembly of single molecules. Nature Nanotech 4, 45–49 (2009). https://doi.org/10.1038/nnano.2008.333

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