title: Digital Microfluidics Assisted Sealing of Individual Magnetic Particles in Femtoliter-Sized Reaction Wells for Single-Molecule Detection.
abstract: Digital microfluidics has emerged in the last years as a promising liquid handling technology for a variety of applications. Here, we describe in detail how to build up an electrowetting-on-dielectric-based digital microfluidic chip with unique advantages for performing single-molecule detection. We illustrate how superparamagnetic particles can be printed with very high loading efficiency (over 98 %) and single-particle resolution in the microwell array patterned in the Teflon-AF®surface of the grounding plate of the chip. Finally, the potential of the device for its application to single-molecule detection is demonstrated by the ultrasensitive detection of the biotinylated enzyme β-Galactosidase captured on streptavidin-coated particles in the described platform.
authors.affiliation: MeBioS - Biosensors, Department of Biosystems, KU Leuven, Willem de Croylaan 42, 3000, Leuven, Belgium.
authors.forename: Deborah
authors.initials: D
authors.lastname: Decrop
authors.affiliation: MeBioS - Biosensors, Department of Biosystems, KU Leuven, Willem de Croylaan 42, 3000, Leuven, Belgium.
authors.forename: Elena Pérez
authors.initials: EP
authors.lastname: Ruiz
authors.affiliation: MeBioS - Biosensors, Department of Biosystems, KU Leuven, Willem de Croylaan 42, 3000, Leuven, Belgium.
authors.forename: Phalguni Tewari
authors.initials: PT
authors.lastname: Kumar
authors.affiliation: MeBioS - Biosensors, Department of Biosystems, KU Leuven, Willem de Croylaan 42, 3000, Leuven, Belgium.
authors.forename: Lisa
authors.initials: L
authors.lastname: Tripodi
authors.affiliation: MeBioS - Biosensors, Department of Biosystems, KU Leuven, Willem de Croylaan 42, 3000, Leuven, Belgium.
authors.forename: Tadej
authors.initials: T
authors.lastname: Kokalj
authors.affiliation: MeBioS - Biosensors, Department of Biosystems, KU Leuven, Willem de Croylaan 42, 3000, Leuven, Belgium. jeroen.lammertyn@kuleuven.be.
authors.forename: Jeroen
authors.initials: J
authors.lastname: Lammertyn
mesh_terms: D004867:Equipment Design; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods*; D044085:Microfluidics / Q000295:instrumentation* / Q000379:methods*; D036103:Nanotechnology; D001616:beta-Galactosidase / Q000378:metabolism
chemical_list: D001616:beta-Galactosidase
keywords: Digital Bioassays; Digital Microfluidics; Magnetic Particles; Microwell Array; Single-Molecule Detection