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Fluorescence and super-resolution standards based on DNA origami

Nature Methods volume 9, pages 1133–1134 (2012)Cite this article

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To the Editor:

In recent years, the ability to validate microscopy techniques as well as distinguish instrument-specific from sample-specific error sources has not kept up with the pace of progress in fluorescence, and especially super-resolution, imaging. Commonly, new methods are demonstrated by imaging cellular filaments whose labeling and preparation cannot be easily reproduced. A desired validation standard requires, on one hand, structural control to position distinct marks at defined distances. On the other hand, the validation standard has to provide stoichiometric control, which implies the ability to place a defined number of molecules of equal brightness per mark. We present molecular rulers based on self-assembled DNA origami structures as a general and highly versatile platform for fluorescence and super-resolution standards. Examples include fluorescence brightness standards as well as standards covering a distance range from 6 to 386 nm that are adapted to the needs of the specific microscopy technique, including stimulated emission depletion (STED), localization-based super-resolution and diffraction-limited microscopy.

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Figure 1: DNA origami standards for fluorescence microscopy.

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Acknowledgements

We thank V. Schüller, R. Schreiber, E. Pibiri, I. Burkhardt, R. Jungmann, P. Nickels, T. Liedl, B. Lalkens and D. Grohmann for help with experiments and fruitful discussions. We are grateful to W. Fouquet, J. Sieber and Leica Microsystems for STED measurements. Financial support by the Biophotonics IV program of the Federal Ministry of Education and Research (BMBF)/Association of German Engineers (VDI)(13N11461) and the German Research Foundation (DFG Ti329/6-1) is gratefully acknowledged.

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  1. Jürgen J Schmied, Andreas Gietl and Phil Holzmeister: These authors contributed equally to this work.

Authors and Affiliations

  1. Institute for Physical and Theoretical Chemistry, Braunschweig University of Technology, Braunschweig, Germany

    Jürgen J Schmied, Andreas Gietl, Phil Holzmeister, Carsten Forthmann, Thorben Dammeyer & Philip Tinnefeld

  2. STS Nanotechnology UG, Freising, Germany

    Christian Steinhauer

Authors
  1. Jürgen J Schmied
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  2. Andreas Gietl
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  3. Phil Holzmeister
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  4. Carsten Forthmann
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  5. Christian Steinhauer
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  6. Thorben Dammeyer
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  7. Philip Tinnefeld
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Correspondence to Philip Tinnefeld.

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Competing interests

Braunschweig University of Technology, which J.J.S., A.G., P.H., C.F., T.D. and P.T. are affiliated with, has filed a provisional patent application for DNA origami–based standards.

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Schmied, J., Gietl, A., Holzmeister, P. et al. Fluorescence and super-resolution standards based on DNA origami. Nat Methods 9, 1133–1134 (2012). https://doi.org/10.1038/nmeth.2254

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