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Nonblinking and long-lasting single-molecule fluorescence imaging

Nature Methods volume 3, pages 891893 (2006) | Download Citation

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Abstract

Photobleaching and blinking of fluorophores pose fundamental limitations on the information content of single-molecule fluorescence measurements. Photoinduced blinking of Cy5 has hampered many previous investigations using this popular fluorophore. Here we show that Trolox in combination with the enzymatic oxygen-scavenging system eliminates Cy5 blinking, dramatically reduces photobleaching and improves the signal linearity at high excitation rates, significantly extending the applicability of single-molecule fluorescence techniques.

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Acknowledgements

We thank J. Liu for introducing Trolox to us and the US National Institutes of Health for grants.

Author information

Affiliations

  1. Department of Physics, Emory University, Atlanta, Georgia 30322, USA.

    • Ivan Rasnik
  2. Department of Physics, University of Illinois, Urbana, Illinois 61801, USA.

    • Sean A McKinney
    •  & Taekjip Ha
  3. Howard Hughes Medical Institute, Urbana, Illinois 61801, USA.

    • Taekjip Ha

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Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Ivan Rasnik.

Supplementary information

PDF files

  1. 1.

    Supplementary Fig. 1

    Single molecule FRET experiments in the presence of 2 mM Trolox or 143 mM BME for the construct of Figure 2d.

  2. 2.

    Supplementary Methods

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DOI

https://doi.org/10.1038/nmeth934

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