Applied Physics B

, Volume 88, Issue 2, pp 161–165

Two-color far-field fluorescence nanoscopy based on photoswitchable emitters

  • H. Bock
  • C. Geisler
  • C.A. Wurm
  • C. von Middendorff
  • S. Jakobs
  • A. Schönle
  • A. Egner
  • S.W. Hell
  • C. Eggeling
Rapid communication

Abstract

We demonstrate two-color far-field fluorescence microscopy with nanoscale spatial resolution based on the photoswitching of individual fluorescent markers. By enabling, recording, and disabling the emission of the reversibly switchable fluorescent protein rsFastLime and of the organic fluorophore cyanine5, we recorded two-color nanoscale images inside whole cells. The position of individual emitters was determined with a typical accuracy of 20 nm, which largely constitutes the lateral resolution of the system. Photoswitching in two-color colocalization experiments represents a major step towards the application of far-field fluorescence nanoscopy to the study of (biological) samples on the macromolecular level.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    E. Abbe, Arch. Mikrosk. Anat. 9, 413 (1873)Google Scholar
  2. 2.
    S.W. Hell, Science 316, 1153 (2007)CrossRefADSGoogle Scholar
  3. 3.
    S.W. Hell, Opt. Commun. 106, 19 (1994)CrossRefADSGoogle Scholar
  4. 4.
    S.W. Hell, J. Wichmann, Opt. Lett. 19, 780 (1994)ADSGoogle Scholar
  5. 5.
    S.W. Hell, S. Jakobs, L. Kastrup, Appl. Phys. A 77, 859 (2003)CrossRefADSGoogle Scholar
  6. 6.
    M. Hofmann, C. Eggeling, S. Jakobs, S.W. Hell, Proc. Nat. Acad. Sci. 102, 17565 (2005)CrossRefADSGoogle Scholar
  7. 7.
    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
  8. 8.
    S.T. Hess, T.P.K. Girirajan, M.D. Mason, Biophys. J. 91, 4258 (2006)CrossRefADSGoogle Scholar
  9. 9.
    M.J. Rust, M. Bates, X. Zhuang, Nature Methods 3, 793 (2006)Google Scholar
  10. 10.
    E. Betzig, Opt. Lett. 20, 237 (1995)ADSGoogle Scholar
  11. 11.
    G.J. Schütz, H. Schindler, T. Schmidt, Biophys. J. 73, 1073 (1997)CrossRefGoogle Scholar
  12. 12.
    T.D. Lacoste, X. Michalet, F. Pinaud, D.S. Chemla, A.P. Alivisatos, S. Weiss, Proc. Nat. Acad. Sci. 97, 9461 (2000)CrossRefADSGoogle Scholar
  13. 13.
    R.E. Thompson, D.R. Larson, W.W. Webb, Biophys. J. 82, 2775 (2002)Google Scholar
  14. 14.
    S.W. Hell, J. Soukka, P.E. Hänninen, Bioimaging 3, 65 (1995)CrossRefGoogle Scholar
  15. 15.
    C. Geisler, A. Schönle, C. von Middendorff, H. Bock, C. Eggeling, A. Egner, S.W. Hell, Appl. Phys. A 88, 223 (2007)CrossRefADSGoogle Scholar
  16. 16.
    E.A. Jares-Erijman, T.M. Jovin, Nature Biotechnol. 21, 1387 (2003)Google Scholar
  17. 17.
    G. Donnert, J. Keller, C.A. Wurm, S.O. Rizzoli, V. Westphal, A. Schönle, R. Jahn, S. Jakobs, C. Eggeling, S.W. Hell, Biophys. J. 92, L67 (2007)CrossRefGoogle Scholar
  18. 18.
    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
  19. 19.
    R. Ando, H. Mizuno, A. Miyawaki, Science 306, 1370 (2004)CrossRefADSGoogle Scholar
  20. 20.
    S. Habuchi, R. Ando, P. Dedecker, W. Verheijen, H. Mizuno, A. Miyawaki, J. Hofkens, Proc. Nat. Acad. Sci. 102, 9511 (2005)CrossRefADSGoogle Scholar
  21. 21.
    M. Bates, T.R. Blosser, X. Zhuang, Phys. Rev. Lett. 94, 108101 (2005)CrossRefADSGoogle Scholar
  22. 22.
    M. Heilemann, E. Margeat, R. Kasper, M. Sauer, P. Tinnefeld, J. Am. Chem. Soc. 127, 3801 (2005)CrossRefGoogle Scholar
  23. 23.
    R. Zondervan, F. Kulzer, M.A. Kolchenko, M. Orrit, J. Phys. Chem. A 108, 1657 (2004)CrossRefGoogle Scholar
  24. 24.
    Z. Huang, D. Ji, A. Xia, Colloid Surf. A 257258, 203 (2005)Google Scholar

Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • H. Bock
    • 1
  • C. Geisler
    • 1
  • C.A. Wurm
    • 1
  • C. von Middendorff
    • 1
  • S. Jakobs
    • 1
  • A. Schönle
    • 1
  • A. Egner
    • 1
  • S.W. Hell
    • 1
  • C. Eggeling
    • 1
  1. 1.Department of NanoBiophotonicsMax Planck Institute for Biophysical ChemistryGöttingenGermany

Personalised recommendations