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Combining optical tweezers, single-molecule fluorescence microscopy, and microfluidics for studies of DNA-protein interactions.

Authors
  • Gross, Peter
  • Farge, Géraldine
  • Peterman, Erwin J G
  • Wuite, Gijs J L
Type
Published Article
Journal
G Protein Coupled Receptors - Structure
Publisher
Elsevier BV
Publication Date
Jan 01, 2010
Volume
475
Pages
427–453
Identifiers
DOI: 10.1016/S0076-6879(10)75017-5
PMID: 20627167
Source
Medline
License
Unknown

Abstract

The technically challenging field of single-molecule biophysics has established itself in the last decade by granting access to detailed information about the fate of individual biomolecules, unattainable in traditional biochemical assays. The appeal of single-molecule methods lies in the directness of the information obtained from individual biomolecules. Technological improvements in single-molecule methods have made it possible to combine optical tweezers, fluorescence microscopy, and microfluidic flow systems. Such a combination of techniques has opened new possibilities to study complex biochemical reactions on the single-molecule level. In this chapter, we provide general considerations for the development of a combined optical trapping, fluorescence microscopy, and microfluidics instrument, along with methods to solve technical issues that are critical for designing successful experiments. Finally, we present several experiments to illustrate the power of this combination of techniques.

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