We have successfully demonstrated selective trapping, concentration, and release of various biological organisms and inert beads by insulator-based dielectrophoresis within a polymeric microfluidic device. The microfluidic channels and internal features, in this case arrays of insulating posts, were initially created through standard wet-etch techniques in glass. This glass chip was then transformed into a nickel stamp through the process of electroplating. The resultant nickel stamp was then used as the replication tool to produce the polymeric devices through injection molding. The polymeric devices were made of Zeonor® 1060R, a polyolefin copolymer resin selected for its superior chemical resistance and optical properties. These devices were then optically aligned with another polymeric substrate that had been machined to form fluidic vias. These two polymeric substrates were then bonded together through thermal diffusion bonding. The sealed devices were utilized to selectively separate and concentrate a variety of biological pathogen simulants and organisms. These organisms include bacteria and spores that were selectively concentrated and released by simply applying D.C. voltages across the plastic replicates via platinum electrodes in inlet and outlet reservoirs. The dielectrophoretic response of the organisms is observed to be a function of the applied electric field and post size, geometry and spacing. Cells were selectively trapped against a background of labeled polystyrene beads and spores to demonstrate that samples of interest can be separated from a diverse background. We have implemented a methodology to determine the concentration factors obtained in these devices.
The ability to detect Weapons of Mass Destruction biological agents rapidly and sensitively is vital to homeland security, spurring development of compact detection systems at Sandia and elsewhere. One such system is Sandia's microseparations-based pChemLab. Many bio-agents are serious health threats even at extremely low concentrations. Therefore, a universal challenge for detection systems is the efficient collection and selective transport of highly diffuse bio-agents against the enormous background of benign particles and species ever present in the ambient environment. We have investigated development of a ''front end'' system for the collection, preconcentration, and selective transport of aerosolized biological agents from dilute (1-10 active particles per liter of air) atmospheric samples, to ultimate concentrations of {approx}20 active particles per microliter of liquid, for interface with microfluidic-based analyses and detection systems. Our approach employs a Sandia-developed aerosol particle-focusing microseparator array to focus size-selected particles into a mating microimpinger array of open microfluidic transport channels. Upon collection (i.e., impingement, submergence, and liquid suspension), microfluidic dielectrophoretic particle concentrators and sorters can be employed to further concentrate and selectively transport bio-agent particles to the sample preparation stages of microfluidic analyses and detection systems. This report documents results in experimental testing, modeling and analysis, component design, and materials fabrication critical to establishing proof-of-principle for this collection ''front end''. Outstanding results have been achieved for the aerodynamic microseparator, and for the post-collection dielectrophoretic concentrator and sorter. Results have been obtained for the microimpinger, too, but issues of particle-trapping by surface tension in liquid surfaces have proven difficult. Subsequent particle submergence into liquid suspension for microfluidic transport has been demonstrated only inefficiently despite significant and varied effort. Importantly, the separate technologies whose development is described, (inertial microseparator, dielectrophoretic corduroy concentrator/sorter) should each, independently, prove greatly useful in a variety of additional applications.
Flow patterns with both recirculating and unidirectional characteristics are useful for controlled mixing and pumping within microfluidic devices. We have developed a fabrication process that converts injection-molded polymer chips into devices that demonstrate induced-charge electroosmosis (ICEO) effects (1,2) in AC fields. Polymeric insulating posts are coated with metal to produce a nonuniform zeta potential under an applied electric field. Induced flows are analyzed by particle image velocimetry. Stable, recirculating flow patterns are discussed, along with their potential to produce well-characterized and reversible streamlines for on-chip mixing in chemical separation and synthesis devices. Asymmetric conductive features can bias the flow direction, generating unidirectional pumping in an AC field. This pumping approach will be discussed in comparison with DC electrokinetic pumps we have studied.
Microfluidic systems are becoming increasingly complicated as the number of applications grows. The use of microfluidic systems for chemical and biological agent detection, for example, requires that a given sample be subjected to many process steps, which requires microvalves to control the position and transport of the sample. Each microfluidic application has its own specific valve requirements and this has precipitated the wide variety of valve designs reported in the literature. Each of these valve designs has its strengths and weaknesses. The strength of the valve design proposed here is its simplicity, which makes it easy to fabricate, easy to actuate, and easy to integrate with a microfluidic system. It can be applied to either gas phase or liquid phase systems. This novel design uses a secondary fluid to stop the flow of the primary fluid in the system. The secondary fluid must be chosen based on the type of flow that it must stop. A dielectric fluid must be used for a liquid phase flow driven by electroosmosis, and a liquid with a large surface tension should be used to stop a gas phase flow driven by a weak pressure differential. Experiments were carried out investigating certain critical functions of the design. These experiments verified that the secondary fluid can be reversibly moved between its 'valve opened' and 'valve closed' positions, where the secondary fluid remained as one contiguous piece during this transport process. The experiments also verified that when Fluorinert is used as the secondary fluid, the valve can break an electric circuit. It was found necessary to apply a hydrophobic coating to the microchannels to stop the primary fluid, an aqueous electrolyte, from wicking past the Fluorinert and short-circuiting the valve. A simple model was used to develop valve designs that could be closed using an electrokinetic pump, and re-opened by simply turning the pump off and allowing capillary forces to push the secondary fluid back into its stowed position.
We have successfully demonstrated selective trapping, concentration, and release of various biological organisms and inert beads by insulator-based dielectrophoresis within a polymeric microfluidic device. The microfluidic channels and internal features, in this case arrays of insulating posts, were initially created through standard wet-etch techniques in glass. This glass chip was then transformed into a nickel stamp through the process of electroplating. The resultant nickel stamp was then used as the replication tool to produce the polymeric devices through injection molding. The polymeric devices were made of Zeonor{reg_sign} 1060R, a polyolefin copolymer resin selected for its superior chemical resistance and optical properties. These devices were then optically aligned with another polymeric substrate that had been machined to form fluidic vias. These two polymeric substrates were then bonded together through thermal diffusion bonding. The sealed devices were utilized to selectively separate and concentrate a biological pathogen simulants. These include spores that were selectively concentrated and released by simply applying D.C. voltages across the plastic replicates via platinum electrodes in inlet and outlet reservoirs. The dielectrophoretic response of the organisms is observed to be a function of the applied electric field and post size, geometry and spacing. Cells were selectively trapped against a background of labeled polystyrene beads and spores to demonstrate that samples of interest can be separated from a diverse background. We have implemented and demonstrated here a methodology to determine the concentration factors obtained in these devices.