An improved surface passivation method for single-molecule studies

Boyang Hua; Kyu Young Han; Ruobo Zhou; Hajin Kim; Xinghua Shi; Sanjaya C Abeysirigunawardena; Ankur Jain; Digvijay Singh; Vasudha Aggarwal; Sarah A Woodson; Taekjip Ha

doi:10.1038/nmeth.3143

An improved surface passivation method for single-molecule studies

Nat Methods. 2014 Dec;11(12):1233-6. doi: 10.1038/nmeth.3143. Epub 2014 Oct 12.

Authors

Affiliations

¹ Center for Biophysics and Computational Biology, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA.
² 1] Department of Physics, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA. [2] Center for the Physics of Living Cells, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA. [3] Howard Hughes Medical Institute, Urbana, Illinois, USA.
³ 1] Department of Physics, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA. [2] Center for the Physics of Living Cells, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA.
⁴ 1] School of Life Sciences, Ulsan National Institute of Science and Technology, Ulsan, Republic of Korea. [2] Center for Soft and Living Matter, Institute for Basic Science, Ulsan, Republic of Korea.
⁵ 1] Howard Hughes Medical Institute, Urbana, Illinois, USA. [2] Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA.
⁶ T.C. Jenkins Department of Biophysics, Johns Hopkins University, Baltimore, Maryland, USA.
⁷ 1] T.C. Jenkins Department of Biophysics, Johns Hopkins University, Baltimore, Maryland, USA. [2] Program in Cell, Molecular, Developmental Biology, and Biophysics (CMDB), Johns Hopkins University, Baltimore, Maryland, USA.
⁸ 1] Center for Biophysics and Computational Biology, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA. [2] Department of Physics, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA. [3] Center for the Physics of Living Cells, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA. [4] Howard Hughes Medical Institute, Urbana, Illinois, USA. [5] Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA.

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.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

DNA-Directed DNA Polymerase / metabolism*
Fluorescence Resonance Energy Transfer / methods*
Humans
Polyethylene Glycols / chemistry*
Polyethylene Glycols / metabolism
Polysorbates / chemistry*
Polysorbates / metabolism
Proliferating Cell Nuclear Antigen / metabolism*
Silanes / chemistry*
Silanes / metabolism
Surface Properties

Substances

Polysorbates
Proliferating Cell Nuclear Antigen
Silanes
Polyethylene Glycols
dichlorodimethylsilane
DNA-Directed DNA Polymerase
POLK protein, human

Abstract

Publication types

MeSH terms

Substances

Grants and funding