Diabetes patients are at risk of having chronic wounds, which would take months to years to resolve naturally. Chronic wounds can be countered using the electrical stimulation technique (EST) by dielectrophoresis (DEP), which is label-free, highly sensitive, and selective for particle trajectory. In this study, we focus on the validation of polystyrene particles of 3.2 and 4.8 μm to predict the behavior of keratinocytes to estimate their crossover frequency (f_XO) using the DEP force (F_DEP) for particle manipulation. MyDEP is a piece of java-based stand-alone software used to consider the dielectric particle response to AC electric fields and analyzes the electrical properties of biological cells. The prototypic 3.2 and 4.8 μm polystyrene particles have f_XO values from MyDEP of 425.02 and 275.37 kHz, respectively. Fibroblast cells were also subjected to numerical analysis because the interaction of keratinocytes and fibroblast cells is essential for wound healing. Consequently, the predicted f_XO from the MyDEP plot for keratinocyte and fibroblast cells are 510.53 and 28.10 MHz, respectively. The finite element method (FEM) is utilized to compute the electric field intensity and particle trajectory based on DEP and drag forces. Moreover, the particle trajectories are quantified in a high and low conductive medium. To justify the simulation, further DEP experiments are carried out by applying a non-uniform electric field to a mixture of different sizes of polystyrene particles and keratinocyte cells, and these results are well agreed. The alive keratinocyte cells exhibit N_DEP force in a highly conductive medium from 100 kHz to 25 MHz. 2D/3D motion analysis software (DIPP-MotionV) can also perform image analysis of keratinocyte cells and evaluate the average speed, acceleration, and trajectory position. The resultant N_DEP force can align the keratinocyte cells in the wound site upon suitable applied frequency. Thus, MyDEP estimates the Clausius–Mossotti factors (CMF), FEM computes the cell trajectory, and the experimental results of prototypic polystyrene particles are well correlated and provide an optimistic response towards keratinocyte cells for rapid wound healing applications.

We present the integration of a flow focusing microfluidic device in a dielectrophoretic application that based on a tapered aluminum microelectrode array (TAMA). The characterization and optimization method of microfluidic geometry performs the hydrodynamic flow focusing on the channel. The sample fluids are hydrodynamically focused into the region of interest (ROI) where the dielectrophoresis force (F_DEP) is dominant. The device geometry is designed using 3D CAD software and fabricated using the micro-milling process combined with soft lithography using PDMS. The flow simulation is achieved using COMSOL Multiphysics 5.5 to study the effect of the flow rate ratio between the sample fluids (Q₁) and the sheath fluids (Q₂) toward the width of flow focusing. Five different flow rate ratios (Q₁/Q₂) are recorded in this experiment, which are 0.2, 0.4, 0.6, 0.8 and 1.0. The width of flow focusing is increased linearly with the flow rate ratio (Q₁/Q₂) for both the simulation and the experiment. At the highest flow rate ratio (Q₁/Q₂ = 1), the width of flow focusing is obtained at 638.66 µm and at the lowest flow rate ratio (Q₁/Q₂ = 0.2), the width of flow focusing is obtained at 226.03 µm. As a result, the flow focusing effect is able to reduce the dispersion of the particles in the microelectrode from 2000 µm to 226.03 µm toward the ROI. The significance of flow focusing on the separation of particles is studied using 10 and 1 µm polystyrene beads by applying a non-uniform electrical field to the TAMA at 10 V_PP, 150 kHz. Ultimately, we are able to manipulate the trajectories of two different types of particles in the channel. For further validation, the focusing of 3.2 µm polystyrene beads within the dominant F_DEP results in an enhanced manipulation efficiency from 20% to 80% in the ROI.

Research relating to dielectrophoresis (DEP) has been progressing rapidly through time as it is a strong and controllable technique for manipulation, separation, preconcentration, and partitioning of protein. Extensive studies have been carried out on protein DEP, especially on Bovine Serum Albumin (BSA). However, these studies involve the usage of dye and fluorescent probes to observe DEP responses as the physical properties of protein albumin molecular structure are translucent. The use of dye and the fluorescent probe could later affect the protein’s physiology. In this article, we review three methods of electrical quantification of DEP responses: electrochemical impedance spectroscopy (EIS), cyclic voltammetry (CV) and capacitance measurement for protein BSA DEP manipulation. The correlation of these methods with DEP responses is further discussed. Based on the observations on capacitance measurement, it can be deduced that the electrical quantifying method is reliable for identifying DEP responses. Further, the possibility of manipulating the protein and electrically quantifying DEP responses while retaining the original physiology of the protein and without the usage of dye or fluorescent probe is discussed.