Automated digital microfluidic platform for magnetic-particle-based immunoassays with optimization by design of experiments.

We introduce an automated digital microfluidic (DMF) platform capable of performing immunoassays from sample to analysis with minimal manual intervention. This platform features (a) a 90 Pogo pin interface for digital microfluidic control, (b) an integrated (and motorized) photomultiplier tube for chemiluminescent detection, and (c) a magnetic lens assembly which focuses magnetic fields into a narrow region on the surface of the DMF device, facilitating up to eight simultaneous digital microfluidic magnetic separations. The new platform was used to implement a three-level full factorial design of experiments (DOE) optimization for thyroid-stimulating hormone immunoassays, varying (1) the analyte concentration, (2) the sample incubation time, and (3) the sample volume, resulting in an optimized protocol that reduced the detection limit and sample incubation time by up to 5-fold and 2-fold, respectively, relative to those from previous work. To our knowledge, this is the first report of a DOE optimization for immunoassays in a microfluidic system of any format. We propose that this new platform paves the way for a benchtop tool that is useful for implementing immunoassays in near-patient settings, including community hospitals, physicians' offices, and small clinical laboratories.

Parallel determination of phenotypic cytotoxicity with a micropattern of mutant cell lines.

This work presents a novel tool, the Continuous Flow Microspotter (CFM) and its use in patterning cellular microarrays of multiple cell types into the bottom of a tissue culture well. The CFM uses a system of isolated microfluidic channels to make an array of localized microspots of adhesion dependent cells in the bottom of a conventional tissue culture well. With this device we have created micropatterns of multiple cell lines in a single tissue culture well and used this system to conduct simultaneous cytotoxicity tests and recover dose survival curves in a parallel study. This mechanism of parallel testing allows the researcher to employ the use of positive and negative controls, as well as compare the chemical response of phenotypes in a tightly controlled microenvironment. For the experiments presented in this paper we have fabricated a CFM with a set of ten microchannels (five inlet channels and five outlet channels) to pattern a row of five microspots consisting of four cellular microspots and one empty spot for background measurements. Micropatterns containing a set of four different Chinese hamster ovarian cell (CHO) mutant phenotypes were deposited into the bottom of commercially available tissue culture wells then interrogated with mitomycin C, a chemotherapeutic agent. This study shows statistically significant (P < 0.05) hypersensitivity of the UV20 CHO mutant to a DNA interstrand cross-linking agent (mitomycin C). Because the CFM is also capable of depositing proteins and other biomolecules to the individual microspots of the array we foresee capabilities of the 48 microspot CFM to multiplex 48 cell types with 48 chemical reagents all within the confines of a 60 mm(2) area.

An integrated optical oxygen sensor fabricated using rapid-prototyping techniques.

This paper details the design and fabrication of an integrated optical biochemical sensor using a select oxygen-sensitive fluorescent dye, tris(2,2'-bipyridyl) dichlororuthenium(ii) hexahydrate, combined with polymeric waveguides that are fabricated on a glass substrate. The sensor uses evanescent interaction of light confined within the waveguide with the dye that is immobilized on an SU-8 waveguide surface. Adhesion of the dye to the integrated waveguide surface is accomplished using a unique process of spin-coating/electrostatic layer-by-layer formation. The SU-8 waveguide was chemically modified to allow the deposition process. Exposure of the dye molecules to the analyte and subsequent chemical interaction is achieved by directly coupling the fluid channel to the integrated waveguide. The completed sensor was linear in the dissolved oxygen across a wide range of interest and had a sensitivity of 0.6 ppm. A unique fabrication aspect of this sensor is the inherent simplicity of the design, and the resulting rapidity of fabrication, while maintaining a high degree of functionality and flexibility.