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Graphene-based metal-induced energy transfer for sub-nanometre optical localization


Single-molecule fluorescence imaging has become an indispensable tool for almost all fields of research, from fundamental physics to the life sciences. Among its most important applications is single-molecule localization super-resolution microscopy (SMLM) (for example, photoactivated localization microscopy (PALM)1, stochastic optical reconstruction microscopy (STORM)2, fluorescent PALM (fPALM)3, direct STORM (dSTORM)4 and point accumulation for imaging in nanoscale topography (PAINT)5), which uses the fact that the centre position of a single molecule’s image can be determined with much higher accuracy than the size of that image itself. However, a big challenge of SMLM is to achieve super-resolution along the third dimension as well. Recently, metal-induced energy transfer (MIET) was introduced to axially localize fluorescent emitters6,7,8,9. This exploits the energy transfer from an excited fluorophore to plasmons in a thin metal film. Here, we show that by using graphene as the ‘metal’ layer, one can increase the localization accuracy of MIET by nearly tenfold. We demonstrate this by axially localizing single emitters and by measuring the thickness of lipid bilayers with ångström accuracy.

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Fig. 1: Graphene-based MIET.
Fig. 2: Axial localization of single molecules with graphene-based MIET.
Fig. 3: Graphene-based MIET measurement of the thickness of SLBs.

Data availability

The data that support the plots within this paper and other findings of this study are available from the corresponding authors upon reasonable request.

Code availability

All Matlab routines and codes used for data analysis of this study are available from the corresponding authors upon request.


  1. 1.

    Betzig, E. et al. Imaging intracellular fluorescent proteins at nanometer resolution. Science 313, 1642–1645 (2006).

    ADS  Google Scholar 

  2. 2.

    Rust, M. J., Bates, M. & Zhuang, X. Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM). Nat. Methods 3, 793–796 (2006).

    Article  Google Scholar 

  3. 3.

    Hess, S. T., Girirajan, T. P. & Mason, M. D. Ultra-high resolution imaging by fluorescence photoactivation localization microscopy. Biophys. J. 91, 4258–4272 (2006).

    ADS  Article  Google Scholar 

  4. 4.

    Heilemann, M. et al. Subdiffraction-resolution fluorescence imaging with conventional fluorescent probes. Angew. Chem. Int. Ed. 47, 6172–6176 (2008).

    Article  Google Scholar 

  5. 5.

    Sharonov, A. & Hochstrasser, R. M. Wide-field subdiffraction imaging by accumulated binding of diffusing probes. Proc. Natl Acad. Sci. USA 103, 18911–18916 (2006).

    ADS  Article  Google Scholar 

  6. 6.

    Chizhik, A. I., Rother, J., Gregor, I., Janshoff, A. & Enderlein, J. Metal-induced energy transfer for live cell nanoscopy. Nat. Photon. 8, 124–127 (2014).

    ADS  Article  Google Scholar 

  7. 7.

    Isbaner, S. et al. Axial colocalization of single molecules with nanometer accuracy using metal-induced energy transfer. Nano Lett. 18, 2616–2622 (2018).

    ADS  Article  Google Scholar 

  8. 8.

    Karedla, N. et al. Single-molecule metal-induced energy transfer (smMIET): resolving nanometer distances at the single-molecule level. ChemPhysChem 15, 705–711 (2014).

    Article  Google Scholar 

  9. 9.

    Karedla, N. et al. Three-dimensional single-molecule localization with nanometer accuracy using metal-induced energy transfer (MIET) imaging. J. Chem. Phys. 148, 204201 (2018).

    ADS  Article  Google Scholar 

  10. 10.

    Huang, B., Wang, W., Bates, M. & Zhuang, X. Three-dimensional super-resolution imaging by stochastic optical reconstruction microscopy. Science 319, 810–813 (2008).

    ADS  Article  Google Scholar 

  11. 11.

    Juette, M. F. et al. Three-dimensional sub-100 nm resolution fluorescence microscopy of thick samples. Nat. Methods 5, 527–529 (2008).

    Article  Google Scholar 

  12. 12.

    Backlund, M. P. et al. Simultaneous, accurate measurement of the 3D position and orientation of single molecules. Proc. Natl Acad. Sci. USA 109, 19087–19092 (2012).

    ADS  Article  Google Scholar 

  13. 13.

    Shtengel, G. et al. Interferometric fluorescent super-resolution microscopy resolves 3D cellular ultrastructure. Proc. Natl Acad. Sci. USA 106, 3125–3130 (2009).

    ADS  Article  Google Scholar 

  14. 14.

    Aquino, D. et al. Two-color nanoscopy of three-dimensional volumes by 4Pi detection of stochastically switched fluorophores. Nat. Methods 8, 353–359 (2011).

    Article  Google Scholar 

  15. 15.

    Bourg, N. et al. Direct optical nanoscopy with axially localized detection. Nat. Photon. 9, 587–593 (2015).

    ADS  Article  Google Scholar 

  16. 16.

    Bon, P. et al. Self-interference 3D super-resolution microscopy for deep tissue investigations. Nat. Methods 15, 449–454 (2018).

    Article  Google Scholar 

  17. 17.

    Novoselov, K. S. et al. Electric field effect in atomically thin carbon films. Science 306, 666–669 (2004).

    ADS  Article  Google Scholar 

  18. 18.

    Wallace, P. R. The band theory of graphite. Phys. Rev. 71, 622 (1947).

    ADS  Article  Google Scholar 

  19. 19.

    Weber, J., Calado, V. & Van De Sanden, M. Optical constants of graphene measured by spectroscopic ellipsometry. Appl. Phys. Lett. 97, 091904 (2010).

    ADS  Article  Google Scholar 

  20. 20.

    Chance, R., Prock, A. & Silbey, R. Molecular fluorescence and energy transfer near interfaces. Adv. Chem Phys. 37, 1–65 (1978).

    Google Scholar 

  21. 21.

    Enderlein, J. Single-molecule fluorescence near a metal layer. Chem. Phys. 247, 1–9 (1999).

    Article  Google Scholar 

  22. 22.

    Chizhik, A. I. et al. Electrodynamic coupling of electric dipole emitters to a fluctuating mode density within a nanocavity. Phys. Rev. Lett. 108, 163002 (2012).

    ADS  Article  Google Scholar 

  23. 23.

    Chizhik, A. I., Gregor, I., Ernst, B. & Enderlein, J. Nanocavity-based determination of absolute values of photoluminescence quantum yields. ChemPhysChem 14, 505–513 (2013).

    Article  Google Scholar 

  24. 24.

    Böhmer, M. & Enderlein, J. Orientation imaging of single molecules by wide-field epifluorescence microscopy. J. Opt. Soc. Am. B 20, 554–559 (2003).

    ADS  Article  Google Scholar 

  25. 25.

    Patra, D., Gregor, I. & Enderlein, J. Image analysis of defocused single-molecule images for three-dimensional molecule orientation studies. J. Phys. Chem. A 108, 6836 (2004).

    Article  Google Scholar 

  26. 26.

    Chizhik, A. I. et al. Probing the radiative transition of single molecules with a tunable microresonator. Nano Lett. 11, 1700–1703 (2011).

    ADS  Article  Google Scholar 

  27. 27.

    Bagatolli, L. A. To see or not to see: lateral organization of biological membranes and fluorescence microscopy. Biochim. Biophys. Acta 1758, 1541–1556 (2006).

    Article  Google Scholar 

  28. 28.

    Schneider, F., Ruhlandt, D., Gregor, I., Enderlein, J. & Chizhik, A. I. Quantum yield measurements of fluorophores in lipid bilayers using a plasmonic nanocavity. J. Phys. Chem. Lett. 8, 1472–1475 (2017).

    Article  Google Scholar 

  29. 29.

    Kučerka, N., Nieh, M.-P. & Katsaras, J. Fluid phase lipid areas and bilayer thicknesses of commonly used phosphatidylcholines as a function of temperature. Biochim. Biophys. Acta 1808, 2761–2771 (2011).

    Article  Google Scholar 

  30. 30.

    Attwood, S. J., Choi, Y. & Leonenko, Z. Preparation of DOPC and DPPC supported planar lipid bilayers for atomic force microscopy and atomic force spectroscopy. Int. J. Mol. Sci. 14, 3514–3539 (2013).

    Article  Google Scholar 

  31. 31.

    Tristram-Nagle, S., Petrache, H. I. & Nagle, J. F. Structure and interactions of fully hydrated dioleoylphosphatidylcholine bilayers. Biophys. J. 75, 917–925 (1998).

    ADS  Article  Google Scholar 

  32. 32.

    Chizhik, A. M. et al. Three-dimensional reconstruction of nuclear envelope architecture using dual-color metal-induced energy transfer imaging. ACS Nano 11, 11839–11846 (2017).

    Article  Google Scholar 

  33. 33.

    Baronsky, T. et al. Cell–substrate dynamics of the epithelial-to-mesenchymal transition. Nano Lett. 17, 3320–3326 (2017).

    ADS  Article  Google Scholar 

  34. 34.

    Simoncelli, S., Makarova, M., Wardley, W. & Owen, D. M. Toward an axial nanoscale ruler for fluorescence microscopy. ACS Nano 11, 11762–11767 (2017).

    Article  Google Scholar 

  35. 35.

    Chizhik, A. M. et al. Dual-color metal-induced and Förster resonance energy transfer for cell nanoscopy. Mol. Biol. Cell 29, 846–851 (2018).

    Article  Google Scholar 

  36. 36.

    Balzarotti, F. et al. Nanometer resolution imaging and tracking of fluorescent molecules with minimal photon fluxes. Science 355, 606–612 (2016).

    ADS  Article  Google Scholar 

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We are grateful to the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) for financial support through projects A06 of SFB 803, A06 of SFB 860, A05 of SFB 937 and through Germany’s Excellence Strategy EXC 2067/1–390729940. We thank the Leibniz Association for financial support through project K76/2017. We also thank B. R. Brueckner for AFM measurements.

Author information




A.G. and N.K. co-designed the project. A.G., A.S. and I.G. performed all the lifetime and defocused imaging measurements. A.G. generated Figs. 2a,c and 3b in the main text. N.K. performed analysis of the defocused imaging data and lifetime data from SLB. A.I.C. carried out the quantum yield measurements with the nanocavity and D.R. performed the corresponding data analysis. S.I. conducted lifetime fitting of single-molecule data and wrote the corresponding section in the Supplementary Information. D.R. helped with writing the MIET routines in Matlab. R.T. designed the DNA origami sample. A.G., N.K. and J.E. carried out the final data analysis, generated all figures (except Figs. 2a,c and 3b) in the main text and wrote the main manuscript. All co-workers were involved with improving the manuscript and writing the Supplementary Information.

Corresponding authors

Correspondence to Narain Karedla or Jörg Enderlein.

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The authors declare no competing interests.

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Supplementary Information

This file contains more information about the work and Supplementary Figs. 1–5.

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Ghosh, A., Sharma, A., Chizhik, A.I. et al. Graphene-based metal-induced energy transfer for sub-nanometre optical localization. Nat. Photonics 13, 860–865 (2019).

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