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Paper based assays are paving the way to automated, simplified, robust and cost-effective point of care testing (POCT). We propose a method for fabricating three dimensional (3D) microfluidic paper based analytical devices (μPADs) via combining thin adhesive films and paper folding, which avoids the use of cellulose powders and the complex folding sequence and simultaneously permits assays in several layers. To demonstrate the effectiveness of this approach, a 3DμPADs was designed to conduct more assays on a small footprint, allowing dual colorimetric and electrochemical detections. More importantly, we further developed a 3D platform for implementing automated and multiplexed ELISA in parallel, since ELISA, a routine and standard laboratory method, has rarely been used in practical analyses outside of the laboratory. In this configuration, complex and multistep diagnostic assays can be carried out with the addition of the sample and buffer in a simple fashion. Using Troponin I as model, the device showed a broad dynamic range of detection with a detection limit of 0.35 ng/mL. Thus, the developed platforms allow for various assays to be cost-effectively carried out on a single 3D device, showing great potential in an academic setting and point of care testing under resource-poor conditions.

Microfluidic systems have been developed widely in scaled-down processes of laboratory techniques, but they are usually limited in achieving stand-alone functionalities. It is highly desirable to exploit an integrated microfluidic device with multiple capabilities such as cell separation, single-cell trapping, and cell manipulation. Herein, we reported a microfluidic platform integrated with actuation electrodes, for separating cells and microbeads, and bipolar electrodes, for trapping, rotating, and propelling single cells and microbeads. The separation of cells and microbeads can be first achieved by deflective dielectrophoresis (DEP) barriers. Trapping experiments with yeast cells and polystyrene (PS) microbeads suspended in aqueous solutions with different conductivities were then conducted, showing that both cells and particles can be trapped at the center of wireless electrodes by negative DEP force. Upon application of a rotating electric field, yeast cells exhibit translational movement along the electrode edges, and self-rotation is seen at an array of bipolar electrodes when electrorotational torque and traveling wave DEP force are applied on the cells. The current approach allows us to switch the propulsion and rotation direction of cells by varying the frequency of the applied electric field. Beyond the achievements of single-cell manipulation, this system permits effective control of several particles or cells simultaneously. The integration of parallel sorting and single trapping stages within a microfluidic chip enables the prospect of high-throughput cell separation, single trapping, and large-scale cell locomotion and rotation in a noninvasive and disposable format, showing great potential in single-cell analysis, targeted drug delivery, and surgery.

We propose a simple, inexpensive microfluidic chip for large-scale trapping of single particles and cells based on induced-charge electroosmosis in a rotating electric field (ROT-ICEO). A central floating electrode array, was placed in the center of the gap between four driving electrodes with a quadrature configuration and used to immobilize single particles or cells. Cells were trapped on the electrode array by the interaction between ROT-ICEO flow and buoyancy flow. We experimentally optimized the efficiency of trapping single particles by investigating important parameters like particle or cell density and electric potential. Experimental and numerical results showed good agreement. The operation of the chip was verified by trapping single polystyrene (PS) microspheres with diameters of 5 and 20 μm and single yeast cells. The highest single particle occupancy of 73% was obtained using a floating electrode array with a diameter of 20 μm with an amplitude voltage of 5 V and frequency of 10 kHz for PS microbeads with a 5-μm diameter and density of 800 particles/μL. The ROT-ICEO flow could hold cells against fluid flows with a rate of less than 0.45 μL/min. This novel, simple, robust method to trap single cells has enormous potential in genetic and metabolic engineering.

We propose a 3D microfluidic mixer based on the alternating current electrothermal (ACET) flow. The ACET vortex is produced by 3D electrodes embedded in the sidewall of the microchannel and is used to stir the fluidic sample throughout the entire channel depth. An optimized geometrical structure of the proposed 3D micromixer device is obtained based on the enhanced theoretical model of ACET flow and natural convection. We quantitatively analyze the flow field driven by the ACET, and a pattern of electrothermal microvortex is visualized by the micro-particle imaging velocimetry. Then, the mixing experiment is conducted using dye solutions with varying solution conductivities. Mixing efficiency can exceed 90% for electrolytes with 0.2 S/m (1 S/m) when the flow rate is 0.364 μL/min (0.728 μL/min) and the imposed peak-to-peak voltage is 52.5 V (35 V). A critical analysis of our micromixer in comparison with different mixer designs using a comparative mixing index is also performed. The ACET micromixer embedded with sidewall 3D electrodes can achieve a highly effective mixing performance and can generate high throughput in the continuous-flow condition.

By increasing the number of floating electrodes or enlarging the width of single floating electrode, this work provides effective ways to strongly improve the particle trapping performance of induced charge electroosmosis (ICEO). Particle trapping with double or triple separate narrow floating electrodes increases the effective actuating range of ICEO flow and therefore enhance the optimum trapping ability to be 1.63 or 2.34 times of that with single narrow electrode (width of L=200μm), and the ideal trapping frequency is independent of the electrode number due to the mutual independence of electrochemical ion relaxation over each electrode. Furthermore, using a single wide floating electrode with the effective width equal to three separate narrow floating electrodes (L=600μm) instead of a single narrow one slightly lowers the ideal trapping frequency due to an increase in the characteristic polarization length, but the trapping performance is only up to 1.59 times of that with original single narrow electrode, implying that vertical channel confinement effect may severely suppresses the effective actuating range of ICEO flow and renders the trapping performance not as expected. Trapping experiments over wide floating electrode with different channel height were carried out, showing that the trapping performance increases by correctly increasing the channel height.

Fluid pumping in microchips using electrokinetic methods has been a hot area of research. This paper mainly investigates effects of DC offset imposed on traveling wave (TW) signal on electroosmotic flow above a spiral electrode array with 800 µm wavelength. The traveling wave voltage with different DC offsets was applied, and four cases were analyzed by superimposing consecutive images. Experiment results indicate that symmetric electrode array energized with DC-biased TW signal can not only generate a prominent improvement in flow rates, but also be capable of altering the flow direction by changing the polarity of electrical signal. Furthermore, such a device can also be used as an effective means to manipulate and separate PS microbeads and cells on their own for very small and non-flowing sample volumes in terms of the combination of the conventional dielectrophoresis (cDEP) forces and traveling wave DEP (twDEP) forces by properly choosing the parameters associated with the Clausius–Mossotti factor (K(w)). Through adjusting the applied frequencies, we successfully separated yeast cells from a mix containing PS microspheres based on the combination of cDEP and twDEP, providing new opportunities for integration with a charge-coupled device for various biomedical diagnostic devices.

Traveling-wave electroosmotic (TWEO) pumping arises from the action of an imposed traveling-wave (TW) electric field on its own induced charge in the diffuse double layer, which is formed on top of an electrode array immersed in electrolyte solutions. Such a traveling field can be merely realized in practice by a discrete electrode array upon which the corresponding voltages of correct phase are imposed. By employing the theory of linear and weakly nonlinear double-layer charging dynamics, a physical model incorporating both the nonlinear surface capacitance of diffuse layer and Faradaic current injection is developed herein in order to quantify the changes in TWEO pumping performance from a single-mode TW to discrete electrode configuration. Benefiting from the linear analysis, we investigate the influence of using discrete electrode array to create the TW signal on the resulting fluid motion, and several approaches are suggested to improve the pumping performance. In the nonlinear regime, our full numerical analysis considering the intervening isolation spacing indicates that a practical four-phase discrete electrode configuration of equal electrode and gap width exhibits stronger nonlinearity than expected from the idealized pump applied with a single-mode TW in terms of voltage-dependence of the ideal pumping frequency and peak flow rate, though it has a much lower pumping performance. For model validation, pumping of electrolytes by TWEO is achieved over a confocal spiral four-phase electrode array covered by an insulating microchannel; measurement of flow velocity indicates the modified nonlinear theory considering moderate Faradaic conductance is indeed a more accurate physical description of TWEO. These results offer useful guidelines for designing high-performance TWEO microfluidic pumps with discrete electrode array.