la800053e_si_001.pdf (34.74 kB)
Fast Mixing and Reaction Initiation Control of Single-Enzyme Kinetics in Confined Volumes
journal contribution
posted on 2008-05-06, 00:00 authored by Seung-Yong Jung, Yu Liu, C. Patrick CollierA 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.