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Published May 6, 2008 | Supplemental Material
Journal Article Open

Fast Mixing and Reaction Initiation Control of Single-Enzyme Kinetics in Confined Volumes

Abstract

A device with femtoliter-scale chambers and controlled reaction initiation was developed for single-molecule enzymology. Initially separated substrate and enzyme streams were rapidly mixed in a microfluidic device and encapsulated in an array of individual microreactors, allowing for enzyme kinetics to be monitored with millisecond dead times and single-molecule sensitivity. Because the arrays of chambers were fabricated by micromolding in PDMS, the chambers were monodisperse in size, and the chamber volume could be systematically controlled. Microreactors could be purged and replenished with fresh reactants for consecutive rounds of observation. Repeated experiments with statistically identical initial conditions could be performed rapidly, with zero cross-talk between chambers in the array.

Additional Information

© 2008 American Chemical Society. Received 7 January 2008. Published online 25 March 2008. Published in print 1 May 2008. The authors thank Alireza Ghaffari at the Micro Nano Fabrication Laboratory at Caltech for help with microfluidic device fabrication.

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August 19, 2023
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