Applied Physics A

, Volume 88, Issue 2, pp 223–226 | Cite as

Resolution of λ /10 in fluorescence microscopy using fast single molecule photo-switching

Invited rapid communication


We demonstrate nanoscale resolution in far-field optical microscopy based on photo-switching of molecules. By enabling, recording and disabling fluorescence from individual labels sequentially, the detection volume is reduced to the size of a single molecule and the diffraction limit is broken. Images of nanostructures milled into a coverslip and tagged by fluorescent proteins could be recorded at 50 nm resolution. Due to the fast and asynchronous image acquisition protocol used in these experiments, we were able to reduce acquisition times to ∼2.5 min, which is two orders of magnitude lower than in previous implementations.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    S.W. Hell, J. Wichmann, Opt. Lett. 19, 780 (1994)ADSCrossRefGoogle Scholar
  2. 2.
    S.W. Hell, S. Jakobs, L. Kastrup, Appl. Phys. A 77, 859 (2003)CrossRefADSGoogle Scholar
  3. 3.
    M. Hofmann, C. Eggeling, S. Jakobs, S.W. Hell, Proc. Nat. Acad. Sci. USA 102, 17565 (2005)CrossRefADSGoogle Scholar
  4. 4.
    R.E. Thompson, D.R. Larson, W.W. Webb, Biophys. J. 82, 2775 (2002)Google Scholar
  5. 5.
    T. Schmidt, G.J. Schutz, W. Baumgartner, H.J. Gruber, H. Schindler, Proc. Nat. Acad. Sci. USA 93, 2926 (1996)CrossRefADSGoogle Scholar
  6. 6.
    A. Yildiz, J.N. Forkey, S.A. McKinney, T. Ha, Y.E. Goldman, P.R. Selvin, Science 300, 2061 (2003)CrossRefADSGoogle Scholar
  7. 7.
    S.W. Hell, J. Soukka, P.E. Hänninen, Bioimaging 3, 65 (1995)CrossRefGoogle Scholar
  8. 8.
    E. Betzig, G.H. Patterson, R. Sougrat, O.W. Lindwasser, S. Olenych, J.S. Bonifacino, M.W. Davidson, J. Lippincott-Schwartz, H.F. Hess, Science 313, 1642 (2006)CrossRefADSGoogle Scholar
  9. 9.
    S.T. Hess, T.P.K. Girirajan, M.D. Mason, Biophys. J. 91, 4258 (2006)CrossRefADSGoogle Scholar
  10. 10.
    M.J. Rust, M. Bates, X. Zhuang, Nat. Methods 3, 793 (2006)Google Scholar
  11. 11.
    W.E. Moerner, Nat. Methods 3, 781 (2006)Google Scholar
  12. 12.
    A.C. Stiel, S. Trowitzsch, G. Weber, M. Andresen, C. Eggeling, S.W. Hell, S. Jakobs, M.C. Wahl, Biochem. J. 402, 35 (2007)CrossRefGoogle Scholar
  13. 13.
    J.A. Högbom, Astrophys. J. Suppl. Ser. 15, 417 (1974)Google Scholar

Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  1. 1.Department of NanoBiophotonicsMax Planck Institute for Biophysical ChemistryGöttingenGermany

Personalised recommendations