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Super-resolution imaging in live Caulobacter crescentus cells using photoswitchable EYFP

Nature Methods volume 5, pages 947–949 (2008)Cite this article

Abstract

The commonly used, monomeric EYFP enabled imaging of intracellular protein structures beyond the optical resolution limit ('super-resolution' imaging) in living cells. By combining photoinduced activation of single EYFP fusions and time-lapse imaging, we obtained sub–40 nm resolution images of the filamentous superstructure of the bacterial actin protein MreB in live Caulobacter crescentus cells. These studies demonstrated that EYFP is a useful emitter for in vivo super-resolution imaging.

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Figure 1: Reactivation of EYFP-MreB fusions in the same live C. crescentus cell.
Figure 2: PALM images of EYFP-MreB in C. crescentus cells.
Figure 3: TL-PALM images of EYFP-MreB in C. crescentus cells showing fewer punctuate spots than continuous-acquisition PALM.

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References

  1. Moerner, W.E. Nat. Methods 3, 781–782 (2006).

    Article  CAS  Google Scholar 

  2. Betzig, E. Opt. Lett. 20, 237–239 (1995).

    Article  CAS  Google Scholar 

  3. Betzig, E. et al. Science 313, 1642–1645 (2006).

    Article  CAS  Google Scholar 

  4. Rust, M.J., Bates, M. & Zhuang, X. Nat. Methods 3, 793–795 (2006).

    Article  CAS  Google Scholar 

  5. Hess, S.T., Girirajan, T.P.K. & Mason, M.D. Biophys. J. 91, 4258–4272 (2006).

    Article  CAS  Google Scholar 

  6. Geisler, C. et al. Appl. Phys. A 88, 223–226 (2007).

    Article  CAS  Google Scholar 

  7. Niu, L. & Yu, P. Biophys. J. 95, 2009–2016 (2008).

    Article  CAS  Google Scholar 

  8. Dickson, R.M., Cubitt, A.B., Tsien, R.Y. & Moerner, W.E. Nature 388, 355–358 (1997).

    Article  CAS  Google Scholar 

  9. Figge, R.M., Divakaruni, A.V. & Gober, J.W. Mol. Microbiol. 51, 1321–1332 (2004).

    Article  CAS  Google Scholar 

  10. Carballido-López, R. Errington, J. Dev. Cell 4, 19–28 (2003).

    Article  Google Scholar 

  11. Gitai, Z., Dye, N. & Shapiro, L. Proc. Natl. Acad. Sci. USA 101, 8643–8648 (2004).

    Article  CAS  Google Scholar 

  12. Kim, S.Y., Gitai, Z., Kinkhabwala, A., Shapiro, L. & Moerner, W.E. Proc. Natl. Acad. Sci. USA 103, 10929–10934 (2006).

    Article  CAS  Google Scholar 

  13. Thanbichler, M. & Shapiro, L. Nat. Rev. Microbiol. 6, 28–40 (2008).

    Article  CAS  Google Scholar 

  14. Thompson, R.E., Larson, D.R. & Webb, W.W. Biophys. J. 82, 2775–2783 (2002).

    Article  CAS  Google Scholar 

  15. Shroff, H., Galbraith, C.G., Galbraith, J.A. & Betzig, E. Nat. Methods 5, 417–423 (2008).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported in part by the US National Institutes of Health grants 1P20HG003638 (Roadmap for Medical Research), 1R01GM085437 (W.E.M.), R01GM051426 (L.S.) and F32GM080008 (postdoctoral fellowship, G.R.B), and a National Science Foundation Graduate Research Fellowship (M.A.T.).

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Authors and Affiliations

  1. Department of Chemistry, Stanford University, 375 North-South Mall, Stanford, 94305, California, USA

    Julie S Biteen, Michael A Thompson, Nicole K Tselentis & W E Moerner

  2. Department of Developmental Biology, Stanford University, B300 Beckman Center, Stanford, 94305, California, USA

    Grant R Bowman & Lucy Shapiro

Authors
  1. Julie S Biteen
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  2. Michael A Thompson
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  3. Nicole K Tselentis
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  4. Grant R Bowman
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  5. Lucy Shapiro
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  6. W E Moerner
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Contributions

J.S.B. and W.E.M. designed experiment, J.S.B. and M.A.T. performed experiments and analyzed data, N.K.T. wrote data acquisition programs, L.S. provided the bacterial strain, G.R.B. sequenced the fusion construct, J.S.B., M.A.T. and W.E.M. wrote the manuscript.

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Correspondence to W E Moerner.

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Supplementary Figures 1–3, Supplementary Methods (PDF 237 kb)

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Biteen, J., Thompson, M., Tselentis, N. et al. Super-resolution imaging in live Caulobacter crescentus cells using photoswitchable EYFP. Nat Methods 5, 947–949 (2008). https://doi.org/10.1038/nmeth.1258

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