Abstract

We report a surface passivation method based on dichlorodimethylsilane (DDS)–Tween-20 for in vitro single-molecule studies, which, under the conditions tested here, more efficiently prevented nonspecific binding of biomolecules than the standard poly(ethylene glycol) surface. The DDS–Tween-20 surface was simple and inexpensive to prepare and did not perturb the behavior and activities of tethered biomolecules. It can also be used for single-molecule imaging in the presence of high concentrations of labeled species in solution.

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Acknowledgements

We thank B. Leslie, S. Syed, T. Ngo, F. Tritschler and Q. Xu for their helpful comments in data analyses and manuscript preparation; J. Guan, C. Yu and S. Granick for the helpful discussions and for generously letting us use the oxygen plasma cleaning machine and Piranha fume hood; T. Le of the H. Kim laboratory at Georgia Institute of Technology and H.R. Koh of the S. Myong laboratory at the University of Illinois for performing independent tests of our protocols; and W. Cheng (University of Michigan) for providing the Alexa 594–labeled protein G B1 domain sample. This work was supported by a US National Science Foundation grant (PHY 1430124 to T.H.) and US National Institutes of Health grants (GM065367 and AG042332 to T.H. and GM60819 to S.A.W.). T.H. is also supported by the Howard Hughes Medical Institute.

Author information

Author notes

    • Ruobo Zhou

    Present address: Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, USA.

Affiliations

  1. Center for Biophysics and Computational Biology, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA.

    • Boyang Hua
    • , Ankur Jain
    • , Digvijay Singh
    • , Vasudha Aggarwal
    •  & Taekjip Ha
  2. Department of Physics, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA.

    • Kyu Young Han
    • , Ruobo Zhou
    •  & Taekjip Ha
  3. Center for the Physics of Living Cells, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA.

    • Kyu Young Han
    • , Ruobo Zhou
    •  & Taekjip Ha
  4. Howard Hughes Medical Institute, Urbana, Illinois, USA.

    • Kyu Young Han
    • , Xinghua Shi
    •  & Taekjip Ha
  5. School of Life Sciences, Ulsan National Institute of Science and Technology, Ulsan, Republic of Korea.

    • Hajin Kim
  6. Center for Soft and Living Matter, Institute for Basic Science, Ulsan, Republic of Korea.

    • Hajin Kim
  7. Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA.

    • Xinghua Shi
    •  & Taekjip Ha
  8. T.C. Jenkins Department of Biophysics, Johns Hopkins University, Baltimore, Maryland, USA.

    • Sanjaya C Abeysirigunawardena
    •  & Sarah A Woodson
  9. Program in Cell, Molecular, Developmental Biology, and Biophysics (CMDB), Johns Hopkins University, Baltimore, Maryland, USA.

    • Sarah A Woodson

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Contributions

B.H. and T.H. designed the project; B.H., K.Y.H., R.Z., H.K. and X.S. performed the experiments; S.C.A. and S.A.W. provided the S4-rRNA complex and three other ribosomal protein samples; B.H., K.Y.H., R.Z., H.K., X.S., A.J., D.S., V.A. and T.H. analyzed the data; and B.H., K.Y.H., R.Z., H.K., X.S. and T.H. wrote the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Taekjip Ha.

Integrated supplementary information

Supplementary information

PDF files

  1. 1.

    Supplementary Text and Figures

    Supplementary Figures 1–14 and Supplementary Protocol

Videos

  1. 1.

    DT20 Cy3 blank channel

    Cy3 background of a blank channel on the DT20 surface with imaging buffer

  2. 2.

    500 nM Atto 647N–labeled ssDNA (18 nt)

    Fluorescence images of nonspecifically bound Atto 647N–labeled ssDNA (18 nt) on the DT20 surface

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DOI

https://doi.org/10.1038/nmeth.3143

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