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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References
Moerner, W.E. Nat. Methods 3, 781–782 (2006).
Betzig, E. Opt. Lett. 20, 237–239 (1995).
Betzig, E. et al. Science 313, 1642–1645 (2006).
Rust, M.J., Bates, M. & Zhuang, X. Nat. Methods 3, 793–795 (2006).
Hess, S.T., Girirajan, T.P.K. & Mason, M.D. Biophys. J. 91, 4258–4272 (2006).
Geisler, C. et al. Appl. Phys. A 88, 223–226 (2007).
Niu, L. & Yu, P. Biophys. J. 95, 2009–2016 (2008).
Dickson, R.M., Cubitt, A.B., Tsien, R.Y. & Moerner, W.E. Nature 388, 355–358 (1997).
Figge, R.M., Divakaruni, A.V. & Gober, J.W. Mol. Microbiol. 51, 1321–1332 (2004).
Carballido-López, R. Errington, J. Dev. Cell 4, 19–28 (2003).
Gitai, Z., Dye, N. & Shapiro, L. Proc. Natl. Acad. Sci. USA 101, 8643–8648 (2004).
Kim, S.Y., Gitai, Z., Kinkhabwala, A., Shapiro, L. & Moerner, W.E. Proc. Natl. Acad. Sci. USA 103, 10929–10934 (2006).
Thanbichler, M. & Shapiro, L. Nat. Rev. Microbiol. 6, 28–40 (2008).
Thompson, R.E., Larson, D.R. & Webb, W.W. Biophys. J. 82, 2775–2783 (2002).
Shroff, H., Galbraith, C.G., Galbraith, J.A. & Betzig, E. Nat. Methods 5, 417–423 (2008).
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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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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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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DOI: https://doi.org/10.1038/nmeth.1258
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