Semi-continuous dielectrophoretic separation at high throughput using printed circuit boards.

Particle separation is an essential part of many processes. One mechanism to separate particles according to size, shape, or material properties is dielectrophoresis (DEP). DEP arises when a polarizable particle is immersed in an inhomogeneous electric field. DEP can attract microparticles toward the local field maxima or repulse them from these locations. In biotechnology and microfluidic devices, this is a well-described and established method to separate (bio-)particles. Increasing the throughput of DEP separators while maintaining their selectivity is a field of current research. In this study, we investigate two approaches to increase the overall throughput of an electrode-based DEP separator that uses selective trapping of particles. We studied how particle concentration affects the separation process by using two differently-sized graphite particles. We showed that concentrations up to 800 mg/L can be processed without decreasing the collection rate depending on the particle size. As a second approach to increase the throughput, parallelization in combination with two four-way valves, relays, and stepper motors was presented and successfully tested to continuously separate conducting from non-conducting particles. By demonstrating possible concentrations and enabling a semi-continuous process, this study brings the low-cost DEP setup based on printed circuit boards one step closer to real-world applications. The principle for semi-continuous processing is also applicable for other DEP devices that use trapping DEP.

Sorting Lithium-Ion Battery Electrode Materials Using Dielectrophoresis.

Lithium-ion batteries (LIBs) are common in everyday life and the demand for their raw materials is increasing. Additionally, spent LIBs should be recycled to achieve a circular economy and supply resources for new LIBs or other products. Especially the recycling of the active material of the electrodes is the focus of current research. Existing approaches for recycling (e.g., pyro-, hydrometallurgy, or flotation) still have their drawbacks, such as the loss of materials, generation of waste, or lack of selectivity. In this study, we test the behavior of commercially available LiFePO4 and two types of graphite microparticles in a dielectrophoretic high-throughput filter. Dielectrophoresis is a volume-dependent electrokinetic force that is commonly used in microfluidics but recently also for applications that focus on enhanced throughput. In our study, graphite particles show significantly higher trapping than LiFePO4 particles. The results indicate that nearly pure fractions of LiFePO4 can be obtained with this technique from a mixture with graphite.

Influence of the filter grain morphology on separation efficiency in dielectrophoretic filtration.

Recovery of noble materials from waste is essential for industries around the globe. Dielectrophoretic (DEP) filtration, an electrically switchable particle separation technique, can be applied to tackle this challenge. It is highly selective regarding particle size, material or shape. Expanding the scope of DEP towards high throughput and improving the trapping efficiency are vital to make DEP a viable robust alternative to conventional separation methods. DEP filtration works by selective immobilisation of particles in a porous medium by the action of an inhomogeneous electric field. The field inhomogeneity comes from scattering an electric field at the phase boundary between the particle suspension and the filter surface. In this article, we show how the filter structure affects the DEP separation. We study fixed bed filters of three different grain types and find that the morphology of the grains highly influences the DEP filter efficiency. Specifically, grains with irregular surface structure and high perceived angularity show high separation efficiency. We believe these insights into the design of DEP filtration will pave the way towards its application in, for example, the recovery of valuable materials from electronic waste dust.

High-throughput dielectrophoretic separator based on printed circuit boards.

The separation of particles with respect to their intrinsic properties is an ongoing task in various fields such as biotechnology and recycling of electronic waste. Especially for small particles in the lower micrometer or nanometer range, separation techniques are a field of current research since many existing approaches lack either throughput or selectivity. Dielectrophoresis (DEP) is a technique that can address multiple particle properties, making it a potential candidate to solve challenging separation tasks. Currently, DEP is mostly used in microfluidic separators and thus limited in throughput. Additionally, DEP setups often require expensive components, such as electrode arrays fabricated in the clean room. Here, we present and characterize a separator based on two inexpensive custom-designed printed circuit boards (80 × 120 mm board size). The boards consist of interdigitated electrode arrays with 250 µ $250\ \umu$ m electrode width and spacing. We demonstrate the separation capabilities using polystyrene particles ranging from 500 nm to 6 µ $6\ \umu$ m in monodisperse experiments. Further, we demonstrate selective trapping at flow rates up to 240 ml/h in the presented device for a binary mixture. Our experiments demonstrate an affordable way to increase throughput in electrode-based DEP separators.

Microfluidic evidence of synergistic effects between mesenchymal stromal cell-derived biochemical factors and biomechanical forces to control endothelial cell function.

A functional vascular system is a prerequisite for bone repair as disturbed angiogenesis often causes non-union. Paracrine factors released from human bone marrow derived mesenchymal stromal cells (BMSCs) have angiogenic effects on endothelial cells. However, whether these paracrine factors participate in blood flow dynamics within bone capillaries remains poorly understood. Here, we used two different microfluidic designs to investigate critical steps during angiogenesis and found pronounced effects of endothelial cell proliferation as well as chemotactic and mechanotactic migration induced by BMSC conditioned medium (CM). The application of BMSC-CM in dynamic cultures demonstrates that bioactive factors in combination with fluidic flow-induced biomechanical signals significantly enhanced endothelial cell migration. Transcriptional analyses of endothelial cells demonstrate the induction of a unique gene expression profile related to tricarboxylic acid cycle and energy metabolism by the combination of BMSC-CM factors and shear stress, which opens an interesting avenue to explore during fracture healing. Our results stress the importance of in vivo - like microenvironments simultaneously including biochemical, biomechanical and oxygen levels when investigating key events during vessel repair. STATEMENT OF SIGNIFICANCE: Our results demonstrate the importance of recapitulating in vivo - like microenvironments when investigating key events during vessel repair. Endothelial cells exhibit enhanced angiogenesis characteristics when simultaneous exposing them to hMSC-CM, mechanical forces and biochemical signals simultaneously. The improved angiogenesis may not only result from the direct effect of growth factors, but also by reprogramming of endothelial cell metabolism. Moreover, with this model we demonstrated a synergistic impact of mechanical forces and biochemical factors on endothelial cell behavior and the expression of genes involved in the TCA cycle and energy metabolism, which opens an interesting new avenue to stimulate angiogenesis during fracture healing.

Microtrap array on a chip for localized electroporation and electro-gene transfection.

We developed a localized single-cell electroporation chip to deliver exogenous biomolecules with high efficiency while maintaining high cell viability. In our microfluidic device, the cells are trapped in a microtrap array by flow, after which target molecules are supplied to the device and electrotransferred to the cells under electric pulses. The system provides the ability to monitor the electrotransfer of exogenous biomolecules in real time. We reveal through numerical simulations that localized electroporation is the mechanism of permeabilization in the microtrap array electroporation device. We demonstrate the simplicity and accuracy of this microtrap technology for electroporation by delivery of both small molecules using propidium iodide and large molecules using plasmid DNA for gene expression, illustrating the potential of this minimally invasive method to be widely used for precise intracellular delivery purposes (from bioprocess engineering to therapeutic applications).

Rational Design and Numerical Analysis of a Hybrid Floating cIDE Separator for Continuous Dielectrophoretic Separation of Microparticles at High Throughput.

Dielectrophoresis (DEP) enables continuous and label-free separation of (bio)microparticles with high sensitivity and selectivity, whereas the low throughput issue greatly confines its clinical application. Herein, we report a novel design of the DEP separator embedded with cylindrical interdigitated electrodes that incorporate hybrid floating electrode layout for (bio)microparticle separation at favorable throughput. To better predict microparticle trajectory in the scaled-up DEP platform, a theoretical model based on coupling of electrostatic, fluid and temperature fields is established, in which the effects of Joule heating-induced electrothermal and buoyancy flows on particles are considered. Size-based fractionation of polystyrene microspheres and dielectric properties-based isolation of MDA-MB-231 from blood cells are numerically realized, respectively, by the proposed separator with sample throughputs up to 2.6 mL/min. Notably, the induced flows can promote DEP discrimination of heterogeneous cells. This work provides a reference on tailoring design of enlarged DEP platforms for highly efficient separation of (bio)samples at high throughput.

Longitudinal Relaxation (T 1) of Methane/Hydrogen Mixtures for Operando Characterization of Gas-Phase Reactions.

Catalytic hydrogenation reactions are important in a modern hydrogen-based society. To optimize these gas-phase reactions, a deep understanding of heat, mass, and momentum transfer inside chemical reactors is required. Nuclear magnetic resonance (NMR) measurements can be used to obtain spatially resolved values of temperature, gas composition, and velocity in the usually opaque catalytic macrostructures. For this, the desired values are calculated from measured NMR parameters like signal amplitude, T 1, or T 2. However, information on how to calculate target values from these NMR parameters in gases is scarce, especially for mixtures of gases. To enable detailed NMR studies of hydrogenation reactions, we investigated the T 1 relaxation of methane and hydrogen, which are two gases commonly present in hydrogenation reactions. To achieve industrially relevant conditions, the temperatures are varied from 290 to 600 K and the pressure from 1 bara to 5 bara, using different mixtures of methane and hydrogen. The results show that hydrogen, which is usually considered to be nondetectable in standard MRI sequences, can be measured at high concentrations, starting at a pressure of 3 bara even at temperatures above 400 K. In the investigated parameter range, the absolute T 1 values of hydrogen show only small dependence on temperature, pressure, and composition, while T 1 of methane is highly dependent on all three parameters. At a pressure of 5 bara, the measured values of T 1 for methane agree very well with theoretical predictions, so that they can also be used for temperature calculations. Further, it can be shown that the same measurement technique can be used to accurately calculate gas ratios inside each voxel. In conclusion, this study covers important aspects of spatially resolved operando NMR measurements of gas-phase properties during hydrogenation reactions at industrially relevant conditions to help improve chemical processes in the gas phase.

Separating microparticles by material and size using dielectrophoretic chromatography with frequency modulation.

Separation of (biological) particles ([Formula: see text]) according to size or other properties is an ongoing challenge in a variety of technical relevant fields. Dielectrophoresis is one method to separate particles according to a diversity of properties, and within the last decades a pool of dielectrophoretic separation techniques has been developed. However, many of them either suffer selectivity or throughput. We use simulation and experiments to investigate retention mechanisms in a novel DEP scheme, namely, frequency-modulated DEP. Results from experiments and simulation show a good agreement for the separation of binary PS particles mixtures with respect to size and more importantly, for the challenging task of separating equally sized microparticles according to surface functionalization alone. The separation with respect to size was performed using 2 [Formula: see text]m and 3 [Formula: see text]m sized particles, whereas separation with respect to surface functionalization was performed with 2 [Formula: see text]m particles. The results from this study can be used to solve challenging separation tasks, for example to separate particles with distributed properties.

A large fixed bed reactor for MRI operando experiments at elevated temperature and pressure.

Recently, in situ studies using nuclear magnetic resonance (NMR) have shown the possibility to monitor local transport phenomena of gas-phase reactions inside opaque structures. Their application to heterogeneously catalyzed reactions remains challenging due to inherent temperature and pressure constraints. In this work, an NMR-compatible reactor was designed, manufactured, and tested, which can endure high temperatures and increased pressure. In temperature and pressure tests, the reactor withstood pressures up to 28 bars at room temperature and temperatures over 400 °C and exhibited only little magnetic shielding. Its applicability was demonstrated by performing the CO2 methanation reaction, which was measured operando for the first time by using a 3D magnetic resonance spectroscopic imaging sequence. The reactor design is described in detail, allowing its easy adaptation for different chemical reactions and other NMR measurements under challenging conditions.

A review of dielectrophoretic separation and classification of non-biological particles.

Dielectrophoresis (DEP) is a selective electrokinetic particle manipulation technology that is applied for almost 100 years and currently finds most applications in biomedical research using microfluidic devices operating at moderate to low throughput. This paper reviews DEP separators capable of high-throughput operation and research addressing separation and analysis of non-biological particle systems. Apart from discussing particle polarization mechanisms, this review summarizes the early applications of DEP for dielectric sorting of minerals and lists contemporary applications in solid/liquid, liquid/liquid, and solid/air separation, for example, DEP filtration or airborne fiber length classification; the review also summarizes developments in DEP fouling suppression, gives a brief overview of electrocoalescence and addresses current problems in high-throughput DEP separation. We aim to provide inspiration for DEP application schemes outside of the biomedical sector, for example, for the recovery of precious metal from scrap or for extraction of metal from low-grade ore.

Aerosol classification by dielectrophoresis: a theoretical study on spherical particles.

The possibilities and limitations using dielectrophoresis (DEP) for the dry classification of spherical aerosol particles was evaluated at low concentrations in a theoretical study. For an instrument with the geometry of concentric cylinders (similar to cylindrical DMA), the dependencies of target particle diameter [Formula: see text], resolution, and yield of the DEP classification on residence time, applied electric field strength, and pressure of the carrier gas were investigated. Further, the diffusion influence on the classification was considered. It was found that [Formula: see text] scales with the mean gas flow velocity [Formula: see text], classifier length L, and electric field strength E as [Formula: see text]. The resolution of the classification depends on the particle diameter and scales proportionally to [Formula: see text]. It is constrained by the flow ratio [Formula: see text] (i.e., sheath gas to aerosol flow), electrode diameters, and applied electric field strength. The classification yield increases with the ratio of the width of the extended outlet slit [Formula: see text] to the diffusion induced broadening [Formula: see text]. As expected, resolution and yield exhibit opposite dependencies on [Formula: see text]. Our simulations show that DEP classification can principally cover a highly interesting particle size range from 100 nm to [Formula: see text] while being directly particle size-selective and particle charge independent.

High-throughput dielectrophoretic filtration of sub-micron and micro particles in macroscopic porous materials.

State-of-the-art dielectrophoretic (DEP) separation techniques provide unique properties to separate particles from a liquid or particles with different properties such as material or morphology from each other. Such separators do not operate at throughput that is sufficient for a vast fraction of separation tasks. This limitation exists because high electric field gradients are required to drive the separation which are generated by electrode microstructures that limit the maximum channel size. Here, we investigate DEP filtration, a technique that uses open porous microstructures instead of microfluidic devices to easily increase the filter cross section and, therefore, also the processable throughput by several orders of magnitude. Previously, we used simple microfluidic porous structures to derive design rules predicting the influence of key parameters on DEP filtration in real complex porous filters. Here, we study in depth DEP filtration in microporous ceramics and underpin the previously postulated dependencies by a broad parameter study (Lorenz et al., 2019). We will further verify our previous claim that the main separation mechanism is indeed positive DEP trapping by showing that we can switch from positive to negative DEP trapping when we increase the electric conductivity of the suspension. Two clearly separated trapping mechanisms (positive and negative DEP trapping) at different conductivities can be observed, and the transition between them matches theoretical predictions. This lays the foundation for selective particle trapping, and the results are a major step towards DEP filtration at high throughput to solve existing separation problems such as scrap recovery or cell separation in liquid biopsy. Graphical abstract.

Polarizability-Dependent Sorting of Microparticles Using Continuous-Flow Dielectrophoretic Chromatography with a Frequency Modulation Method.

The separation of microparticles with respect to different properties such as size and material is a research field of great interest. Dielectrophoresis, a phenomenon that is capable of addressing multiple particle properties at once, can be used to perform a chromatographic separation. However, the selectivity of current dielectrophoretic particle chromatography (DPC) techniques is limited. Here, we show a new approach for DPC based on differences in the dielectrophoretic mobilities and the crossover frequencies of polystyrene particles. Both differences are addressed by modulating the frequency of the electric field to generate positive and negative dielectrophoretic movement to achieve multiple trap-and-release cycles of the particles. A chromatographic separation of different particle sizes revealed the voltage dependency of this method. Additionally, we showed the frequency bandwidth influence on separation using one example. The DPC method developed was tested with model particles, but offers possibilities to separate a broad range of plastic and metal microparticles or cells and to overcome currently existing limitations in selectivity.

Bridging the scales in high-throughput dielectrophoretic (bio-)particle separation in porous media.

Dielectrophoresis (DEP) is a versatile technique for the solution of difficult (bio-)particle separation tasks based on size and material. Particle motion by DEP requires a highly inhomogeneous electric field. Thus, the throughput of classical DEP devices is limited by restrictions on the channel size to achieve large enough gradients. Here, we investigate dielectrophoretic filtration, in which channel size and separation performance are decoupled because particles are trapped at induced field maxima in a porous separation matrix. By simulating microfluidic model porous media, we derive design rules for DEP filters and verify them using model particles (polystyrene) and biological cells (S. cerevisiae, yeast). Further, we bridge the throughput gap by separating yeast in an alumina sponge and show that the design rules are equally applicable in real porous media at high throughput. While maintaining almost 100% efficiency, we process up to 9 mL min-1, several orders of magnitude more than most state-of-the-art DEP applications. Our microfluidic approach provides new insight into trapping dynamics in porous media, which even can be applied in real sponges. These results pave the way toward high-throughput retention, which is capable of solving existing problems such as cell separation in liquid biopsy or precious metal recovery.

Influence of geometry and material of insulating posts on particle trapping using positive dielectrophoresis.

Insulator-based dielectrophoresis (iDEP) is a powerful particle analysis technique based on electric field scattering at material boundaries which can be used, for example, for particle filtration or to achieve chromatographic separation. Typical devices consist of microchannels containing an array of posts but large scale application was also successfully tested. Distribution and magnitude of the generated field gradients and thus the possibility to trap particles depends apart from the applied field strength on the material combination between post and surrounding medium and on the boundary shape. In this study we simulate trajectories of singe particles under the influence of positive DEP that are flowing past one single post due to an external fluid flow. We analyze the influence of key parameters (excitatory field strength, fluid flow velocity, particle size, distance from the post, post size, and cross-sectional geometry) on two benchmark criteria, i.e., a critical initial distance from the post so that trapping still occurs (at fixed particle size) and a critical minimum particle size necessary for trapping (at fixed initial distance). Our approach is fundamental and not based on finding an optimal geometry of insulating structures but rather aims to understand the underlying phenomena of particle trapping. A sensitivity analysis reveals that electric field strength and particle size have the same impact, as have fluid flow velocity and post dimension. Compared to these parameters the geometry of the post's cross-section (i.e. rhomboidal or elliptical with varying width-to-height or aspect ratio) has a rather small influence but can be used to optimize the trapping efficiency at a specific distance. We hence found an ideal aspect ratio for trapping for each base geometry and initial distance to the tip which is independent of the other parameters. As a result we present design criteria which we believe to be a valuable addition to the existing literature.

Electrodeless dielectrophoresis: Impact of geometry and material on obstacle polarization.

Insulator-based (electrodeless) dielectrophoresis (iDEP) is a promising particle manipulation technique, based on movement of matter in inhomogeneous fields. The inhomogeneity of the field arises because the excitatory field distorts at obstacles (posts). This effect is caused by accumulation of polarization charges at material interfaces. In this study, we utilize a multipole expansion method to investigate the influence of geometry and material on field distortion of posts with arbitrary cross-sections in homogeneous electric fields applied perpendicular to the longitudinal axis of the post. The post then develops a multipole parallel or anti parallel to the excitatory field. The multipoles intensity is defined by the post's structure and material properties and directly influences the DEP particle trapping potential. We analyzed posts with circular and rhombus-shaped cross-sections with different cross-sectional width-to-height ratios and permittivities for their polarization intensity, multipole position, and their particle trapping behavior. A trade-off between high maximum field gradient and high coverage range of the gradient is presented, which is determined by the sharpness of the post's edges. We contribute to the overall understanding of the post polarization mechanism and expect that the results presented will help optimizing the structure of microchannels with arrays of posts for electrodeless DEP application.