Experimental insights into electrocatalytic [Cp*Rh(bpy)Cl]+ mediated NADH regeneration.

NADH plays a crucial role in many enzymatically catalysed reactions. Due to the high costs of NADH a regeneration mechanism of this cofactor can enlarge the applications of enzymatic reactions dramatically. This paper gives a thorough system analysis of the mediated electrochemical regeneration of active NADH using cyclic voltammograms and potentiostatic measurements with varying pH, electrode potential, and electrolyte solution, highlighting the system's limiting conditions, elucidating optimal working parameters for the electrochemical reduction of NAD+, and bringing new insight on the oxidation of inactive reduction products. Using [Cp*Rh(bpy)Cl]+ as an electron mediator dramatically increases the percentage of enzymatically active electrochemically reduced NADH from 15% (direct) to 99% (mediated) with a faradaic efficiency of up to 86%. Furthermore, investigations of the catalytic mechanisms of [Cp*Rh(bpy)Cl]+ clarifies the necessary conditions for its functioning and questions the proposed reaction mechanism by two-step reduction where first the mediator is reduced and then brought in contact with NAD+.

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.

Influence of electrode potential, pH and NAD+ concentration on the electrochemical NADH regeneration.

Electrochemical NAD+ reduction is a promising method to regenerate NADH for enzymatic reactions. Many different electrocatalysts have been tested in the search for high yields of the 1,4-isomer of NADH, the active NADH, but aside from electrode material, other system parameters such as pH, electrode potential and educt concentration also play a role in NADH regeneration. The effect of these last three parameters and the mechanisms behind their influence on NADH regeneration was systematically studied and presented in this paper. With percentages of active NADH ranging from 10 to 70% and faradaic efficiencies between 1 and 30%, it is clear that all three system parameters drastically affect the reaction outcome. As a proof of principle, the NAD+ reduction in the presence of pyruvate and lactate dehydrogenase was performed. It could be shown that the electrochemical NADH regeneration can also be done successfully in parallel to enzymatically usage of the regenerated cofactor.

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.

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.

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.

Descriptors for High Throughput in Structural Materials Development.

The development of novel structural materials with increasing mechanical requirements is a very resource-intense process if conventional methods are used. While there are high-throughput methods for the development of functional materials, this is not the case for structural materials. Their mechanical properties are determined by their microstructure, so that increased sample volumes are needed. Furthermore, new short-time characterization techniques are required for individual samples which do not necessarily measure the desired material properties, but descriptors which can later be mapped on material properties. While universal micro-hardness testing is being commonly used, it is limited in its capability to measure sample volumes which contain a characteristic microstructure. We propose to use alternative and fast deformation techniques for spherical micro-samples in combination with classical characterization techniques such as XRD, DSC or micro magnetic methods, which deliver descriptors for the microstructural state.

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.

Applying Alkyl-Chain Surface Functionalizations in Mesoporous Inorganic Structures: Their Impact on Gas Flow and Selectivity Depending on Temperature.

Porous inorganic capillary membranes are prepared to serve as model structures for the experimental investigation of the gas transport in functionalized mesopores. The porous structures possess a mean pore diameter of 23 nm which is slightly reduced to 20 nm after immobilizing C16-alkyl chains on the surface. Gas permeation measurements are performed at temperatures ranging from 0 to 80 °C using Ar, N2, and CO2. Nonfunctionalized structures feature a gas transport according to Knudsen diffusion with regard to gas flow and selectivity. After C16-functionalization, the gas flow is reduced by a factor of 10, and the ideal selectivities deviate from the Knudsen theory. CO2 adsorption measurements show a decrease in total amount of adsorbed gas and isosteric heat of adsorption. It is hypothesized that the immobilized C16-chains sterically influence the gas transport behavior without a contribution from adsorption effects. The reduced gas flow derives from an additional surface resistance caused by the C16-chains spacially limiting the adsorption and desorption directions for gas molecules propagating through the structure, resulting in longer diffusion paths. In agreement, the gas flow is found to correlate with the molecular diameter of the gas species (CO2 < Ar < N2) increasing the resistance for larger molecules. This affects the ideal selectivities with the relation [Formula: see text]. The influence on selectivity increases with increasing temperature which leads to the conclusion that the temperature induced movement of the C16-chains is responsible for the stronger interaction between gas molecules and surface functional groups.

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.

A fouling suppression system in submerged membrane bioreactors using dielectrophoretic forces.

A novel method was developed to suppress membrane fouling in submerged membrane bioreactors. The method is based on the dielectrophoretic (DEP) motion of particles in an inhomogeneous electrical field. Using a real sample of biomass as feed, the fouling-suppression performance using DEP with different electrical field intensities (60-160 V) and different frequencies (50-1000 Hz) was investigated. The fouling-suppression performance was found to relate closely with the intensity and frequency of the electrical field. A stronger electrical field was found to better recover the filtrate flux. This is because of a stronger DEP force acting on the biomass particles close to the membrane's surface. Above an intensity and frequency value of 130 V and 1 kHz, respectively the permeate flux was reduced due to an electrothermal effect.