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Single-molecule measurements of transient biomolecular complexes through microfluidic dilution

Journal Article


Abstract


  • Single-molecule confocal microscopy experiments require concentrations which are low enough to guarantee that, on average, less than one single molecule resides in the probe volume at any given time. Such concentrations are, however, significantly lower than the dissociation constants of many biological complexes which can therefore dissociate under single-molecule conditions. To address the challenge of observing weakly bound complexes in single-molecule experiments in solution, we have designed a microfluidic device that rapidly dilutes samples by up to one hundred thousand times, allowing the observation of unstable complexes before they dissociate. The device can interface with standard biochemistry laboratory experiments and generates a spatially uniform dilution that is stable over time allowing the quantification of the relative concentrations of different molecular species.

UOW Authors


  •   Kjaergaard, Magnus (external author)
  •   Rajah, Luke (external author)
  •   Jönsson, Peter (external author)
  •   Vendruscolo, Michele (external author)
  •   Knowles, Tuomas P. J. (external author)
  •   Klenerman, David (external author)
  •   Horrocks, Mathew H. (external author)

Publication Date


  • 2013

Citation


  • Horrocks, M. H., Rajah, L., Jönsson, P., Kjaergaard, M., Vendruscolo, M., Knowles, T. P. J. & Klenerman, D. (2013). Single-molecule measurements of transient biomolecular complexes through microfluidic dilution. Analytical Chemistry, 85 (14), 6855-6859.

Scopus Eid


  • 2-s2.0-84880525663

Ro Metadata Url


  • http://ro.uow.edu.au/smhpapers/3358

Has Global Citation Frequency


Number Of Pages


  • 4

Start Page


  • 6855

End Page


  • 6859

Volume


  • 85

Issue


  • 14

Abstract


  • Single-molecule confocal microscopy experiments require concentrations which are low enough to guarantee that, on average, less than one single molecule resides in the probe volume at any given time. Such concentrations are, however, significantly lower than the dissociation constants of many biological complexes which can therefore dissociate under single-molecule conditions. To address the challenge of observing weakly bound complexes in single-molecule experiments in solution, we have designed a microfluidic device that rapidly dilutes samples by up to one hundred thousand times, allowing the observation of unstable complexes before they dissociate. The device can interface with standard biochemistry laboratory experiments and generates a spatially uniform dilution that is stable over time allowing the quantification of the relative concentrations of different molecular species.

UOW Authors


  •   Kjaergaard, Magnus (external author)
  •   Rajah, Luke (external author)
  •   Jönsson, Peter (external author)
  •   Vendruscolo, Michele (external author)
  •   Knowles, Tuomas P. J. (external author)
  •   Klenerman, David (external author)
  •   Horrocks, Mathew H. (external author)

Publication Date


  • 2013

Citation


  • Horrocks, M. H., Rajah, L., Jönsson, P., Kjaergaard, M., Vendruscolo, M., Knowles, T. P. J. & Klenerman, D. (2013). Single-molecule measurements of transient biomolecular complexes through microfluidic dilution. Analytical Chemistry, 85 (14), 6855-6859.

Scopus Eid


  • 2-s2.0-84880525663

Ro Metadata Url


  • http://ro.uow.edu.au/smhpapers/3358

Has Global Citation Frequency


Number Of Pages


  • 4

Start Page


  • 6855

End Page


  • 6859

Volume


  • 85

Issue


  • 14