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Published March 2011 | Supplemental Material
Journal Article Open

High-throughput single-molecule optofluidic analysis

Abstract

We describe a high-throughput, automated single-molecule measurement system, equipped with microfluidics. The microfluidic mixing device has integrated valves and pumps to accurately accomplish titration of biomolecules with picoliter resolution. We demonstrate that the approach enabled rapid sampling of biomolecule conformational landscape and of enzymatic activity, in the form of transcription by Escherichia coli RNA polymerase, as a function of the chemical environment.

Additional Information

© 2011 Nature Publishing Group. Received 09 July 2010; Accepted12 January 2011; Published online 06 February 2011. We thank R. Colyer, P. Blainey and other members of the Weiss and Quake laboratories for helpful discussions. This work was supported by US National Science Foundation Frontiers in Integrative Biological Research grant 0623664 and National Institutes of Health grant GM069709. Fluorescence spectroscopy was performed at the University of California, Los Angeles and California NanoSystems Institute Advanced Light Microscopy and Spectroscopy Shared Facility. A.M.S. was supported by the Stanford University Diversifying Academia, Recruiting Excellence fellowship. Author Contributions: S.K., A.M.S. and D.S.M. designed experiments, conducted experiments, wrote and implemented data acquisition and analysis software, and analyzed data. R.R.L. analyzed data. S.K., A.M.S., S.R.Q., S.W. and D.S.M. assisted in writing and editing of the manuscript.

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Supplemental Material - nmeth.1569-S2.zip

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August 22, 2023
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