University of Washington

Boston University

Anson Hatch

Paul Yager

Andrew Kamholz

Bernhard Hans Weigl

Charles Cantor

Takeshi Sano

Karl F Bohringer

Catherine R Cabrera

Matthew S Munson

Karl B&ouml;hringer

Berhard Weigl

John Misasi

Chandran R Sabanayagam

Methods and apparatuses are provided for determining presence and concentration of analytes by exploiting molecular binding reactions and differential diffusion rates. Analyte particles and binding particles are allowed to diffuse toward each other, and slowing of the diffusion front is detected when they meet. From the position of the diffusion front, presence and concentration of analyte particles can be determined. One embodiment provides a competitive immunoassay in a microfluidic format. This diffusion immunoassay (DIA) relies on measuring the concentration of labeled antigen along one dimension of a microchannel after allowing it to diffuse for a short time into a region containing specific antibodies. A simple microfluidic device, the T-Sensor, was used to implement a DIA to measure the concentration of phenytoin, a small drug molecule. Concentrations of analyte over the range of 50 to 1600 nM can be measured in less than a minute. The assay is homogeneous, rapid, requires only microliter volumes of reagents and sample, and is applicable to a wide range of analytes, including therapeutic drugs, molecular biological markers, and environmental contaminants. Methods for separating particles of similar size in a diffusion separator are also provided.

Microscale diffusion immunoassay

This invention provides methods for using liquid junction potentials to control the transport of charged particles in fluid streams that are in laminar flow within microfluidic channels. Applications of the methods of this invention include sample preconditioning (removal of interfering substances), electrophoretic separation (fractionation) of charged particles, enhanced or delayed mixing of charged particles across a fluid interface relative to diffusion only, focusing charged particles in a fluid stream in one or two dimensions, and concentration of charged reactants at a fluid interface.

Use of liquid junction potentials for electrophoresis without applied voltage in a microfluidic channel

Magnetically actuated fluid handling devices using magnetic fluid to move one or more fluids (gases or liquids or both) through microsized flow channels are provided. Fluid handling devices include micropumps and microvalves. Magnetically actuated slugs of magnetic fluid are moved within microchannels of a microfluidic device to facilitate valving and/or pumping of fluids and no separate pump is required. The magnets used to control fluid movement can be either individual magnets moved along the flow channels or one or more arrays of magnets whose elements can be individually controlled to hold or move a magnetic slug. Fluid handling devices include those having an array of electromagnets positioned along a flow channel which are turned on and off in a predetermined pattern to move magnetic fluid slugs in desired paths in the flow channel. The fluid handling devices of the present invention can handle gases and liquids simultaneously and thus can be made to be self-priming. These devices are more resistant to fluctuations in fluid input than other types of micropumps which need to be tuned to pump either liquid or gas. In a particular embodiment, a micropump having a loop channel containing a stationary magnetic slug and one or more moving slugs is provided.

Magnetically actuated fluid handling devices for microfluidic applications

The present invention generally relates to high density nucleic acid arrays and methods of synthesizing nucleic acid sequences on a solid surface. Specifically, the present invention contemplates the use of stabilized nucleic acid primer sequences immobilized on solid surfaces, and circular nucleic acid sequence templates combined with the use of isothermal rolling circle amplification to thereby increase nucleic acid sequence concentrations in a sample or on an array of nucleic acid sequences.

Nucleic acid arrays and methods of synthesis

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Seattle, WA, United States of America

Anson Hatch