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Single-molecule measurements of transient biomolecular complexes through microfluidic dilution

Horrocks, Mathew H ; Rajah, Luke ; Jönsson, Peter LU ; Kjaergaard, Magnus ; Vendruscolo, Michele ; Knowles, Tuomas P J and Klenerman, David (2013) In Analytical Chemistry 85(14). p.9-6855
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... (More)

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.

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author
; ; ; ; ; and
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Indicator Dilution Techniques, Microfluidic Analytical Techniques, Microscopy, Fluorescence, Oligonucleotides, Journal Article, Research Support, Non-U.S. Gov't
in
Analytical Chemistry
volume
85
issue
14
pages
5 pages
publisher
The American Chemical Society (ACS)
external identifiers
  • pmid:23782428
  • scopus:84880525663
ISSN
1520-6882
DOI
10.1021/ac4010875
language
English
LU publication?
no
id
54c2cba3-c1f2-485c-9868-57035907e7ec
date added to LUP
2018-01-26 10:28:42
date last changed
2021-05-05 01:23:34
@article{54c2cba3-c1f2-485c-9868-57035907e7ec,
  abstract     = {<p>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.</p>},
  author       = {Horrocks, Mathew H and Rajah, Luke and Jönsson, Peter and Kjaergaard, Magnus and Vendruscolo, Michele and Knowles, Tuomas P J and Klenerman, David},
  issn         = {1520-6882},
  language     = {eng},
  month        = {07},
  number       = {14},
  pages        = {9--6855},
  publisher    = {The American Chemical Society (ACS)},
  series       = {Analytical Chemistry},
  title        = {Single-molecule measurements of transient biomolecular complexes through microfluidic dilution},
  url          = {http://dx.doi.org/10.1021/ac4010875},
  doi          = {10.1021/ac4010875},
  volume       = {85},
  year         = {2013},
}