Design of micromagnetic arrays for on-chip separation of superparamagnetic bead aggregates and detection of a model protein and double-stranded DNA analytes.

Magnetically actuated lab-on-a-chip (LOC) technologies have enabled rapid, highly efficient separation of specific biomarkers and cells from complex biological samples. Nonlinear magnetophoresis (NLM) is a technique that uses a microfabricated magnet array (MMA) and a time varying external magnetic field to precisely control the transport of superparamagnetic (SPM) beads on the surface of a chip based on their size and magnetization. We analyze the transport and separation behavior of SPM monomers and dimers on four MMA geometries, i.e., circular, triangular, square and rectangular shaped micromagnets, across a range of external magnetic field rotation frequencies. The measured critical frequency of the SPM beads on an MMA, i.e., the velocity for which the hydrodynamic drag on a bead exceeds the magnetic force, is closely related to the local magnetic flux density landscape on a micromagnet in the presence of an external magnetic field. A set of design criteria has been established for the optimization of MMAs for NLM separation, with particular focus on the shape of the micromagnets forming the array. The square MMA was used to detect a model protein biomarker and gene fragment based on a magnetic bead assembly (MBA) assay. This assay uses ligand functionalized SPM beads to capture and directly detect an analyte through the formation of SPM bead aggregates. These beads aggregates were detected through NLM separation and microscopic analysis resulting in a highly sensitive assay that did not use carrier fluid.

Optical detection of the magnetophoretic transport of superparamagnetic beads on a micromagnetic array.

Micromagnetic arrays (MMAs) have proven to be powerful tools for controlling the transport and separation of bioanalytes, i.e., they allow bioanalyte-superparamagnetic (SPM) bead complexes of specific size and magnetization to be moved in a synchronized manner that is precisely controlled with the orientation of an external magnetic field. This article presents a laser-photodetector system for the simple detection of individual SPM beads moving on a specific region of an MMA. This system detects the SPM beads through the change in intensity of reflective light as they move from the highly reflective micromagnetics to the supporting substrate. We demonstrate that this opti-MMA system allowed the size, number, and magnetic and optical properties of the SPM beads to be rapidly determined for regions > 49 µm2 in size. The response of the opti-MMA system was characterized in several optical configurations to develop a theoretical description of its sensitivity and dynamic range. The speed, low-cost, and sensitivity of this system promises to allow MMAs to be readily applied in in vitro diagnostics and biosensing.

Direct identification of the herpes simplex virus UL27 gene through single particle manipulation and optical detection using a micromagnetic array.

Magnetophoretic lab on a chip technologies are rapidly evolving into integrated systems for the identification of biomarkers and cells with ultra-high sensitivity. We demonstrate the highly efficient detection of the Human herpes simplex virus type 1 (HSV) UL27 gene through the programmed assembly of superparamagnetic (SPM) nanoparticles based on oligonucleotide hybridization. The state of assembly of the SPM nanoparticles was determined by optical signature of the synchronized motion on the beads on a micromagnetic array (MMA). This technique has been used to identify <200 copies of the HSV UL27 gene without amplification in less than 20 minutes. The MAA can also be used to separate gene-SPM bead aggregates from millions of unreacted SPM beads based on nonlinear magnetophoresis (NLM). The MMA-optical detection system promises to enable highly sensitive, nucleic acid analysis to be performed without amplification and with the consumption of minimal amounts of reagent.

A microfluidic dual gradient generator for conducting cell-based drug combination assays.

We present a microfluidic chip that generates linear concentration gradients of multiple solutes that are orthogonally-aligned to each other. The kinetics of gradient formation was characterized using a fluorescent tracer matching the molecular weight of small inhibitory drugs. Live-cell signalling and motility experiments were conducted to demonstrate the potential uses and advantages of the device. A431 epidermoid carcinoma cells, where EGF induces apoptosis in a concentration-dependent manner, were simultaneously exposed to gradients of MEK inhibitor and EGF receptor (EGFR) inhibitor. By monitoring live caspase activation in the entire chip, we were able to quickly assess the combinatorial interaction between MEK and EGFR pathways, which otherwise would require costly and time consuming titration experiments. We also characterized the motility and morphology of MDA-MB-231 breast cancer cells exposed to orthogonal gradients of EGF and EGFR inhibitor. The microfluidic chip not only permitted the quantitative analysis of a population of cells exposed to drug combinations, but also enabled the morphological characterization of individual cells. In summary, our microfluidic device, capable of establishing concentration gradients of multiple compounds over a group of cells, facilitates and accelerates in vitro cell biology experiments, such as those required for cell-based drug combination assays.