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Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM)

Nature Methods 3, 793–796 (2006) | Download Citation

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Abstract

We have developed a high-resolution fluorescence microscopy method based on high-accuracy localization of photoswitchable fluorophores. In each imaging cycle, only a fraction of the fluorophores were turned on, allowing their positions to be determined with nanometer accuracy. The fluorophore positions obtained from a series of imaging cycles were used to reconstruct the overall image. We demonstrated an imaging resolution of 20 nm. This technique can, in principle, reach molecular-scale resolution.

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Acknowledgements

This work is supported by in part by the US National Institutes of Health, the Defense Advance Research Projects Agency and a Packard Science and Engineering Fellowship (to X.Z.). X.Z. is a Howard Hughes Medical Institute Investigator.

Author information

    • Michael J Rust
    •  & Mark Bates

    These authors contributed equally to this work.

Affiliations

  1. Department of Physics, Harvard University, 12 Oxford Street, Cambridge, Massachusetts 02138, USA.

    • Michael J Rust
    •  & Xiaowei Zhuang

  2. Division of Engineering and Applied Sciences, Harvard University, 12 Oxford Street, Cambridge, Massachusetts 02138, USA.

    • Mark Bates

  3. Department of Chemistry and Chemical Biology, Harvard University, 12 Oxford Street, Cambridge, Massachusetts 02138, USA.

    • Xiaowei Zhuang

  4. Howard Hughes Medical Institute, Harvard University, 12 Oxford Street, Cambridge, Massachusetts 02138, USA.

    • Xiaowei Zhuang

Authors

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Contributions

M.J.R. and M.B. conceived the STORM imaging concept. M.J.R., DNA imaging and data analysis; M.B., RecA imaging and data acquisition. X.Z. supervised the project.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Xiaowei Zhuang.

Supplementary information

PDF files

  1. 1.

    Supplementary Fig. 1

    Switching of Cy3-Cy5–labeled antibody.

  2. 2.

    Supplementary Fig. 2

    Localization accuracy of single switches before and after drift correction.

  3. 3.

    Supplementary Fig. 3

    Fluorescence time trace of DNA labeled with multiple switches over many STORM imaging cycles.

  4. 4.

    Supplementary Methods

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DOI

https://doi.org/10.1038/nmeth929