title: A prototype microfluidic platform for miniaturization and automation of serial dilution and dose-response assays.
abstract: A novel microfluidic device was designed and developed to miniaturize, multiplex, and automate serial dilution and three-reagent dose-response assays using submicroliter quantities of reagents. This prototype microfluidic device can be used to measure enzyme kinetics and to test a chemical lead's response to a target by fluorescent readout using common plate readers and detection systems. The prototype microfluidic system yielded serial dilution and dose-response assay data comparable to results obtained from manual titrations and reagent additions performed using a microwell plate. Enzyme kinetics were highly reproducible using these devices, although Michaelis-Menten kinetics results differed from those obtained in the microwell plate. In all cases reported here, assays performed on the microfluidic format required lower volumes of reagents compared with the microwell plate. In addition to savings in reagent consumption, the microfluidic devices and bench-top instruments offer additional advantages over conventional solutions, including a small footprint and compatibility with commercially available fluorescence detectors. Future directions for the prototype technology are discussed.
authors.affiliation: Nanostream, Pasadena, CA 91107, USA. jeff.koehler@nanostream.com
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Koehler
authors.forename: Surekha
authors.initials: S
authors.lastname: Vajjhala
authors.forename: Courtney
authors.initials: C
authors.lastname: Coyne
authors.forename: Terence
authors.initials: T
authors.lastname: Flynn
authors.forename: Marci
authors.initials: M
authors.lastname: Pezzuto
authors.forename: Manda
authors.initials: M
authors.lastname: Williams
authors.forename: Leanna
authors.initials: L
authors.lastname: Levine
mesh_terms: D001331:Automation; D001681:Biological Assay / Q000295:instrumentation* / Q000379:methods; D003287:Contrast Media; D004305:Dose-Response Relationship, Drug; D004798:Enzymes / Q000378:metabolism; D019793:Fluorescein; D019788:Fluorodeoxyglucose F18 / Q000494:pharmacology; D007091:Image Processing, Computer-Assisted; D007201:Indicator Dilution Techniques / Q000295:instrumentation*; D007700:Kinetics; D008838:Microcomputers; D036103:Nanotechnology; D010865:Pilot Projects; D019275:Radiopharmaceuticals / Q000494:pharmacology; D001616:beta-Galactosidase / Q000037:antagonists & inhibitors
chemical_list: D003287:Contrast Media; D004798:Enzymes; D019275:Radiopharmaceuticals; D019788:Fluorodeoxyglucose F18; D001616:beta-Galactosidase; D019793:Fluorescein