title: Single-enzyme analysis in a droplet-based micro- and nanofluidic system.
abstract: The kinetic activity of individual enzyme molecules was determined in aqueous droplets generated in a nano- and microfluidic device. To avoid high background noise, the enzyme and substrate solution was confined into femtoliter carriers, achieving high product concentrations from single-molecule encapsulation. The tiny droplets (φ ~ 2.5-3 μm) generated from this fluidic system were highly monodisperse, beneficial for an analysis of single enzyme activity. The method presented here allows to follow large numbers of individual droplets over time. The instrumental requirements are furthermore modest, since the small droplet size allows to use of standard microscope and standard Pyrex glass chips as well as the use of relatively high enzyme concentrations (nM range) for single molecule encapsulation.
authors.affiliation: BIOS/Lab-on-chip group, MESA+ Institute for Nanotechnology, University of Twente, Enschede, The Netherlands.
authors.forename: Rerngchai
authors.initials: R
authors.lastname: Arayanarakool
authors.forename: Lingling
authors.initials: L
authors.lastname: Shui
authors.forename: Servé W M
authors.initials: SW
authors.lastname: Kengen
authors.forename: Albert
authors.initials: A
authors.lastname: van den Berg
authors.forename: Jan C T
authors.initials: JC
authors.lastname: Eijkel
mesh_terms: D019793:Fluorescein / Q000737:chemistry; D007700:Kinetics; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation / Q000379:methods*; D036103:Nanotechnology / Q000295:instrumentation / Q000379:methods*; D011435:Propyl Gallate / Q000737:chemistry; D013050:Spectrometry, Fluorescence; D013379:Substrate Specificity; D001616:beta-Galactosidase / Q000032:analysis*
chemical_list: D011435:Propyl Gallate; D001616:beta-Galactosidase; D019793:Fluorescein