Helical dielectrophoretic particle separator fabricated by conformal spindle printing

This paper reports the fabrication and testing of a helical cell separator that uses insulator-based dielectrophoresis as the driving force of its separation. The helical channel shape’s main advantage is its constant curvature radius which generates a constant electric field gradient. The presented separator was fabricated by extruding a sacrificial ink on rotating spindles using a computer- controlled robot. After being assembled, connected to the reservoir and encapsulated in epoxy resin, the ink was removed to create a helical microchannel. The resulting device was tested by circulating polystyrene microbeads of 4 and 10 μm diameter through its channel using a voltage of 900 VDC. The particles were separated with efficiencies of 94.0% and 92.5%, respectively. However, roughness in some parts of the channel and connections that had larger diameters compared to the channel created local electric field gradients which, doubtless, hindered separation. It is a promising device that could lead the way toward portable and affordable medical devices

Micromachines for Dielectrophoresis

An outstanding compilation that reflects the state-of-the art on Dielectrophoresis (DEP) in 2020. Contributions include: - A novel mathematical framework to analyze particle dynamics inside a circular arc microchannel using computational modeling. - A fundamental study of the passive focusing of particles in ratchet microchannels using direct-current DEP. - A novel molecular version of the Clausius-Mossotti factor that bridges the gap between theory and experiments in DEP of proteins. - The use of titanium electrodes to rapidly enrich T. brucei parasites towards a diagnostic assay. - Leveraging induced-charge electrophoresis (ICEP) to control the direction and speed of Janus particles. - An integrated device for the isolation, retrieval, and off-chip recovery of single cells. - Feasibility of using well-established CMOS processes to fabricate DEP devices. - The use of an exponential function to drive electrowetting displays to reduce flicker and improve the static display performance. - A novel waveform to drive electrophoretic displays with improved display quality and reduced flicker intensity. - Review of how combining electrode structures, single or multiple field magnitudes and/or frequencies, as well as variations in the media suspending the particles can improve the sensitivity of DEP-based particle separations. - Improvement of dielectrophoretic particle chromatography (DPC) of latex particles by exploiting differences in both their DEP mobility and their crossover frequencies

Micro/Nano-Chip Electrokinetics

Micro/nanofluidic chips have found increasing applications in the analysis of chemical and biological samples over the past two decades. Electrokinetics has become the method of choice in these micro/nano-chips for transporting, manipulating and sensing ions, (bio)molecules, fluids and (bio)particles, etc., due to the high maneuverability, scalability, sensitivity, and integrability. The involved phenomena, which cover electroosmosis, electrophoresis, dielectrophoresis, electrohydrodynamics, electrothermal flow, diffusioosmosis, diffusiophoresis, streaming potential, current, etc., arise from either the inherent or the induced surface charge on the solid-liquid interface under DC and/or AC electric fields. To review the state-of-the-art of micro/nanochip electrokinetics, we welcome, in this Special Issue of Micromachines, all original research or review articles on the fundamentals and applications of the variety of electrokinetic phenomena in both microfluidic and nanofluidic devices

Dielectrophoresis: An Approach to Increase Sensitivity, Reduce Response Time and to Suppress Nonspecific Binding in Biosensors?

The performance of receptor-based biosensors is often limited by either diffusion of the analyte causing unreasonable long assay times or a lack of specificity limiting the sensitivity due to the noise of nonspecific binding. Alternating current (AC) electrokinetics and its effect on biosensing is an increasing field of research dedicated to address this issue and can improve mass transfer of the analyte by electrothermal effects, electroosmosis, or dielectrophoresis (DEP). Accordingly, several works have shown improved sensitivity and lowered assay times by order of magnitude thanks to the improved mass transfer with these techniques. To realize high sensitivity in real samples with realistic sample matrix avoiding nonspecific binding is critical and the improved mass transfer should ideally be specific to the target analyte. In this paper we cover recent approaches to combine biosensors with DEP, which is the AC kinetic approach with the highest selectivity. We conclude that while associated with many challenges, for several applications the approach could be beneficial, especially if more work is dedicated to minimizing nonspecific bindings, for which DEP offers interesting perspectives

Dielectrophoretic nanotweezers for single-particle force spectroscopy

Studying the structural and functional properties of biological molecules using single-molecule techniques has been fundamental in establishing a comprehensive understanding of the mechanisms that govern complex living systems. The aim of this thesis was to develop a dielectrophoretic nanotweezers setup to provide a new approach for applying and measuring dielectrophoretic forces on individual particles in aqueous solution. To achieve this goal, dual-barrel quartz nanopipettes were filled with pyrolytic carbon, forming a pair of nanoelectrodes at their tip. Their size and shape were characterised both through SEM imaging and electrochemical cyclic voltammetry. By applying low AC voltages (< 1 V) between these nanoelectrodes, very strong and highly inhomogeneous electric fields were generated at the nanopipette’s tip to form dielectrophoretic nanotweezers. The frequency of the electric field was tuned so that individual entities were either attracted or repelled depending on their dielectric properties. In this work, experimental measurements of the dielectrophoretic force acting on single polystyrene beads (2 μm) were acquired by single particle tracking on an inverted fluorescence microscope. The spatial coordinates of individual beads were extracted from their trajectories as a function of time under trapping conditions. The force magnitude, measured from their velocity over distance from the nanotweezers tip, was found in the femtonewtons range for a set of applied voltages and frequencies. In addition, the electric field distribution was simulated close to the nanotweezers tip by a finite element model developed for this system. Estimations of the dielectrophoretic force magnitude for different nanotweezers geometries were also performed. Overall, the simple operational mechanism and design of these dielectrophoretic nanotweezers combined with their ability to be controlled in three-dimensions, make them a versatile and promising platform for single-particle manipulation and force probing

Isomotive dielectrophoresis for enhanced analyses of cell subpopulations.

As the relentless dream of creating a true lab-on-a-chip device is closer to realization than ever before, which will be enabled through efficient and reliable sample characterization systems. Dielectrophoresis (DEP) is a term used to describe the motion of dielectric particles/ cells, by means of a non-uniform electric field (AC or DC). Cells of different dielectric properties (i.e., size, interior properties, and membrane properties) will act differently under the influence of dielectrophoretic force. Therefore, DEP can be used as a powerful, robust, and flexible tool for cellular manipulation, separation, characterization, and patterning. However, most recent DEP applications focus on trapping, separation, or sorting particles. The true value of DEP lies in its analytical capabilities which can be achieved by utilizing isomotive dielectrophoresis (isoDEP). In isoDEP, the gradient of the electric field-squared is constant, hence, upon the application of electric field, all particles/cells that share the same dielectric properties will feel the same constant dielectrophoretic force i.e., translate through the micro-channel at the same velocity. However, DEP is not the only acting force upon particles inside an isoDEP device, other electrokinetics, including but not limited to electrothermal hydrodynamics, might act on particles simultaneously. Within this dissertation, electrothermal-based experiments have been conducted to assess the effect of such undesired forces. Also, to maximize the relative DEP force over other forces for a given cell/particle size, design parameters such as microchannel width, height, fabrication materials, lid thickness, and applied electric field must be properly tuned. In this work, scaling law analyses were developed to derive design rules that relate those tunable parameters to achieve the desired dielectrophoretic force for cell analysis. Initial results indicated that for a particle suspended in 10 mS/m media, if the channel width and height are below 10 particle diameters, the electrothermal-driven flow is reduced by ∼ 500 times compared to the 500 µm thick conventional isoDEP device. Also, Replacing glass with silicon as the device’s base for an insulative-based isoDEP, reduces the electrothermal induced flow by ∼ 20 times. Within this dissertation, different device designs and fabrication methods were attempted in order to achieve an isoDEP platform that can characterize and differentiate between live and dead phytoplankton cells suspended in the same solution. Unfortunately, unwanted electrokinetics (predicted by the previously mentioned scaling law analysis) prevented comprehensive isoDEP analysis of phytoplankton cells. Due to isoDEP device limitations and other complications, other techniques were pursued to electrically characterize phytoplankton cells in suspension. An electrochemical-based platform utilizing impedance spectroscopy measurements was used to extract the electrical properties of phytoplankton cells in suspension. Impedance spectroscopy spectra were acquired, and the single-shell model was applied to extract the specific membrane capacitance, cytoplasm permittivity, and conductivity of assumingly spherical cells in suspension utilizing Maxwell’s mixture theory of a controlled volume fraction of cells. The impedance of suspensions of algae were measured at different frequencies ranging from 3 kHz to 10 MHz and impedance values were compared to investigate differences between two types of cells by characterizing their change in cytoplasm permittivity and membrane capacitance. Differentiation between healthy control and nitrogen-depleted cultured algae was attempted. The extracted specific membrane capacitances of Chlamydomonas and Selenastrum were 15:57 ± 3:62 and 40:64 ± 12:6 mF/m2 respectively. Successful differentiation based on the specific membrane capacitance of different algae species was achieved. However, no significant difference was noticed between nitrogen abundant and nitrogen depleted cultures. To investigate the potential of isoDEP for cell analysis, a comparison to existing dielectrophoresis-based electrokinetic techniques was encouraged, including electrorotation (ROT) microfluidic platforms. The ROT microfluidic chip was used to characterize M17, HEK293, T-lymphocytes, and Hela single cells. Through hands-on experience with ROT, the advantages and disadvantages of this approach and isoDEP are apparent. IsoDEP proves to be a good characterization tool for subpopulation cell analysis with potential higher throughput compared to ROT while maintaining simple fabrication and operation processes. To emphasize the role of dielectrophoresis in biology, further studies utilizing the 3DEP analytical system were used to determine the electrical properties of Drosophila melanogaster (Kc167) cells ectopically expressing Late embryogenesis abundant (LEA) proteins from the anhydrobiotic brine shrimp, Artemia franciscana. Dielectrophoretic-based characterization data demonstrates that single expression of two different LEA proteins, AfrLEA3m and AfrLEA6, both increase cytoplasmic conductivity of Kc167 cells to a similar extend above control values. The extracted DEP data supported previously reported data suggesting that AfrLEA3m can interact directly with membranes during water stress. This hypothesis was strengthened using scanning electron microscopy, where cells expressing AfrLEA3m were found to retain their spherical morphology during desiccation, while control cells exhibited a larger variety of shapes in the desiccated state

Microdevices and Microsystems for Cell Manipulation

Microfabricated devices and systems capable of micromanipulation are well-suited for the manipulation of cells. These technologies are capable of a variety of functions, including cell trapping, cell sorting, cell culturing, and cell surgery, often at single-cell or sub-cellular resolution. These functionalities are achieved through a variety of mechanisms, including mechanical, electrical, magnetic, optical, and thermal forces. The operations that these microdevices and microsystems enable are relevant to many areas of biomedical research, including tissue engineering, cellular therapeutics, drug discovery, and diagnostics. This Special Issue will highlight recent advances in the field of cellular manipulation. Technologies capable of parallel single-cell manipulation are of special interest

Design of a dielectrophoretic cell loading device

In recent years there has been an increasing interest in studying individual cells, and structures that physically entrap one or few cells have been developed for this purpose, but the approaches to load cells into these structures leave a lot to be desired. This dissertation discusses the design of a device that loads cells suspended in a solution into microvials using a combination of dielectrophoresis and fluid flow, which offers significant advantages over previous loading approaches. The basic concept is to use fluid flow and dielectrophoretic forces to position a given cell above a given vial, within an array of similar vials, and then bringing the cell into the vial. The loading of several cells flowing in a channel into a vial in a matter of seconds is demonstrated. The design of the loading device spurred the development of novel topics in the area of dielectrophoresis. The structures into which cells are loaded produce "parasitic cages". The effect of multiple electric fields and at multiple frequencies had to be explored to eliminate the parasitic cages, and new theory was developed to describe the phenomenon in a straight forward and convenient way. The design process of dielectrophoretic structures known as flow through sorters was simplified significantly using a method that relies on non dimensional analysis and a figure of merit. These topics investigated have broader applications than just loading cells into vials. The dissertation demonstrates technologies and design and fabrication methods key to the cell loading design. The dissertation ends by describing the design of a device that can be implemented to load cells into vials on integrated circuit chips and outlining this device's expected characteristics and performance based on the theory and methods presented through the dissertation

Microfluidic device with 3D electrode structure for high throughput dielectrophoretic applications

Cataloged from PDF version of thesis.Includes bibliographical references (leaves 55-62).Thesis (M.S.): Bilkent University, The Department of Mechanical Engineering, İhsan Doğramacı Bilkent University, 2014.Microfluidics is the combination of micro/nano fabrication techniques together with knowledge of fluid behavior at the microscopic level to pursue powerful techniques in controlling, manipulating and measuring chemical, physical and biological processes at micro/nano scale. Sorting and separation of bio-particles are highly considered in diagnostics and biological analyses. By implementing the characteristics of microscale flow phenomenon, dielectrophoresis (DEP) has offered unique advantages for microfluidic devices. In DEP devices asymmetric pair of planar or three dimensional (3D) electrodes could be employed to generate non-uniform electric field. In DEP applications, facing 3D sidewall electrodes is considered to be the key solution of increasing device throughput because of producing homogeneous electric fields along the height of microchannels. Despite all advantages, fabrication of 3D vertical electrodes requires considerable challenge. In this thesis, in order to highlight the advantage of 3D electrodes over planar electrodes, the simulations are performed. Based on the developed computational model, the design parameters are decided. For the fabrication of the device, two different fabrication techniques have been proposed. In the first method, both the mold and the electrodes are fabricated using high precision machining. In the second method, the mold is fabricated with tilted sidewalls using high precision machining and the electrodes are deposited on the sidewall using sputtering together with a shadow mask fabricated using wire electric discharge machining (WEDM). The both techniques are assessed as highly repeatable and robust methods. Only the manipulation of particles with negative-DEP has been demonstrated in the experiments, and the throughput values up to 105 particles/min have been reached in a continuous flow.by Soheila Zeinali.M.S