Image-based sorting and negative dielectrophoresis for high purity cell and particle separation.

Microelectrode arrays are used to sort single fluorescently labeled cells and particles as they flow through a microfluidic channel using dielectrophoresis. Negative dielectrophoresis is used to create a "Dielectrophoretic virtual channel" that runs along the center of the microfluidic channel. By switching the polarity of the electrodes, the virtual channel can be dynamically reconfigured to direct particles along a different path. This is demonstrated by sorting particles into two microfluidic outlets, controlled by an automated system that interprets video data from a color camera and makes complex sorting decisions based on color, intensity, size, and shape. This enables the rejection of particle aggregates and other impurities, and the system is optimized to isolate high purity populations from a heterogeneous sample. Green beads are isolated from an excess of red beads with 100% purity at a rate of up to 0.9 particles per second, in addition application to the sorting of osteosarcoma and human bone marrow cells is evidenced. The extension of Dielectrophoretic Virtual Channels to an arbitrary number of sorting outputs is examined, with design, simulation, and experimental verification of two alternate geometries presented and compared.

Lab-on-chip measurement of nitrate and nitrite for in situ analysis of natural waters.

Microfluidic technology permits the miniaturization of chemical analytical methods that are traditionally undertaken using benchtop equipment in the laboratory environment. When applied to environmental monitoring, these "lab-on-chip" systems could allow high-performance chemical analysis methods to be performed in situ over distributed sensor networks with large numbers of measurement nodes. Here we present the first of a new generation of microfluidic chemical analysis systems with sufficient analytical performance and robustness for deployment in natural waters. The system detects nitrate and nitrite (up to 350 µM, 21.7 mg/L as NO(3)(-)) with a limit of detection (LOD) of 0.025 µM for nitrate (0.0016 mg/L as NO(3)(-)) and 0.02 µM for nitrite (0.00092 mg/L as NO(2)(-)). This performance is suitable for almost all natural waters (apart from the oligotrophic open ocean), and the device was deployed in an estuarine environment (Southampton Water) to monitor nitrate+nitrite concentrations in waters of varying salinity. The system was able to track changes in the nitrate-salinity relationship of estuarine waters due to increased river flow after a period of high rainfall. Laboratory characterization and deployment data are presented, demonstrating the ability of the system to acquire data with high temporal resolution.

Negative DEP traps for single cell immobilisation.

We present a novel design of micron-sized particle trap that uses negative dielectrophoresis (nDEP) to trap cells in high conductivity physiological media. The design is scalable and suitable for trapping large numbers of single cells. Each trap has one electrical connection and the design can be extended to produce a large array. The trap consists of a metal ring electrode and a surrounding ground plane, which create a closed electric field cage in the centre. The operation of the device was demonstrated by trapping single latex spheres and HeLa cells against a moving fluid. The dielectrophoretic holding force was determined experimentally by measuring the displacement of a trapped particle in a moving fluid. This was then compared with theory by numerically solving the electric field for the electrodes and calculating the trapping force, demonstrating good agreement. Analysis of the 80 microm diameter trap showed that a 15.6 microm diameter latex particle could be held with a force of 23 pN at an applied voltage of 5 V peak-peak.

Trapping single human osteoblast-like cells from a heterogeneous population using a dielectrophoretic microfluidic device.

We describe a system for the isolation, concentration, separation, and recovery of human osteoblast-like cells from a heterogeneous population using dielectrophoretic ring traps. Cells flowing in a microfluidic channel are immobilized inside an electric field cage using negative dielectrophoresis. A planar ring electrode creates a closed trap while repelling surrounding cells. Target cells are identified by fluorescent labeling, and are trapped as they pass across a ring electrode by an automated system. We demonstrate recovery of small populations of human osteoblast-like cells with a purity of 100%, which in turn demonstrates the potential of such a device for cell selection from a heterogeneous population.