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Sharper low-power STED nanoscopy by time gating


Applying pulsed excitation together with time-gated detection improves the fluorescence on-off contrast in continuous-wave stimulated emission depletion (CW-STED) microscopy, thus revealing finer details in fixed and living cells using moderate light intensities. This method also enables super-resolution fluorescence correlation spectroscopy with CW-STED beams, as demonstrated by quantifying the dynamics of labeled lipid molecules in the plasma membrane of living cells.

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Figure 1: Principle of g-STED.
Figure 2: g-STED fluorescence nanoscopy.
Figure 3: g-STED-FCS.


  1. Hell, S.W. & Wichmann, J. Opt. Lett. 19, 780–782 (1994).

    Article  CAS  Google Scholar 

  2. Klar, T.A., Jakobs, S., Dyba, M., Egner, A. & Hell, S.W. Proc. Natl. Acad. Sci. USA 97, 8206–8210 (2000).

    Article  CAS  Google Scholar 

  3. Hell, S.W. Nat. Methods 6, 24–32 (2009).

    Article  CAS  Google Scholar 

  4. Eggeling, C. et al. Nature 457, 1159–1162 (2009).

    Article  CAS  Google Scholar 

  5. Willig, K.I., Harke, B., Medda, R. & Hell, S.W. Nat. Methods 4, 915–918 (2007).

    Article  CAS  Google Scholar 

  6. Moneron, G. et al. Opt. Express 18, 1302–1309 (2010).

    Article  CAS  Google Scholar 

  7. Donnert, G. et al. Proc. Natl. Acad. Sci. USA 103, 11440–11445 (2006).

    Article  CAS  Google Scholar 

  8. Leutenegger, M., Eggeling, C. & Hell, S.W. Opt. Express 18, 26417–26429 (2010).

    Article  CAS  Google Scholar 

  9. Schrader, M. et al. Bioimaging 3, 147–153 (1995).

    Article  CAS  Google Scholar 

  10. Westphal, V. & Hell, S.W. Phys. Rev. Lett. 94, 143903 (2005).

    Article  Google Scholar 

  11. Auksorius, E. et al. Opt. Lett. 33, 113–115 (2008).

    Article  Google Scholar 

  12. Hell, S.W., Jakobs, S. & Kastrup, L. Appl. Phys., A Mater. Sci. Process. 77, 859–860 (2003).

    Article  CAS  Google Scholar 

  13. Ringemann, C. et al. N. J. Phys. 11, 103054 (2009).

    Article  Google Scholar 

  14. Han, K.Y. et al. Nano Lett. 9, 3323–3329 (2009).

    Article  CAS  Google Scholar 

  15. Jelezko, F. & Wrachtrup, J. Phys. Status Solidi A 203, 3207–3225 (2006).

    Article  CAS  Google Scholar 

  16. Griesbeck, O., Baird, G.S., Campbell, R.E., Zacharias, D.A. & Tsien, R.Y. J. Biol. Chem. 276, 29188–29194 (2001).

    Article  CAS  Google Scholar 

  17. Lamesch, P. et al. Genomics 89, 307–315 (2007).

    Article  CAS  PubMed  Google Scholar 

  18. Chiantia, S., Ries, J., Kahya, N. & Schwille, P. ChemPhysChem 7, 2409–2418 (2006).

    Article  CAS  Google Scholar 

  19. Widengren, J., Mets, U. & Rigler, R. J. Phys. Chem. 99, 13368–13379 (1995).

    Article  CAS  Google Scholar 

  20. Zander, C. et al. Appl. Phys. B 63, 517–523 (1996).

    Article  CAS  Google Scholar 

  21. Maus, M. et al. Anal. Chem. 73, 2078–2086 (2001).

    Article  CAS  Google Scholar 

  22. Cotlet, M. et al. J. Phys. Chem. B 105, 4999–5006 (2001).

    Article  CAS  Google Scholar 

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We thank A. Schönle and M. Leutenegger for fruitful discussions, A. Schönle for support with the software Imspector, V. Müller and A. Honigmann for support with the FCS measurements, and U. Gemm for support with the electronics. C. Wurm, T. Gilat and E. Rothermel helped prepare samples.

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Authors and Affiliations



G.V., G.M., J.E., C.E. and S.W.H. conceived and designed the study. G.V. performed theoretical studies. V.W. designed electronic components. G.V., G.M., K.Y.H., H.T. and M.R. performed experiments. G.V., G.M., K.Y.H. and C.E. analyzed data. G.V., G.M., C.E. and S.W.H. wrote the manuscript. All authors discussed the conceptual and practical implications of the method at all stages.

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Correspondence to Stefan W Hell.

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Competing interests

G.V., G.M., K.Y.H, V.W., M.R., J.E., C.E. and S.W.H. have filed a patent application on the method presented.

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Supplementary Figures 1–8 and Supplementary Note 1 (PDF 4079 kb)

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Vicidomini, G., Moneron, G., Han, K. et al. Sharper low-power STED nanoscopy by time gating. Nat Methods 8, 571–573 (2011).

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