Visual and Quantitative Analysis of the Trapping Volume in Dielectrophoresis of Nanoparticles.

Nanoparticle manipulation requires careful analysis of the forces at play. Unfortunately, traditional force measurement techniques based on the particle velocity do not provide sufficient resolution, while balancing approaches involving counteracting forces are often cumbersome. Here, we demonstrate that a nanoparticle dielectrophoretic response can be quantitatively studied by a straightforward visual delineation of the dielectrophoretic trapping volume. We reveal this volume by detecting the width of the region depleted of gold nanoparticles by the dielectrophoretic force. Comparison of the measured widths for various nanoparticle sizes with numerical simulations obtained by solving the particle-conservation equation shows excellent agreement, thus providing access to the particle physical properties, such as polarizability and size. These findings can be further extended to investigate various types of nano-objects, including bio- and molecular aggregates, and offer a robust characterization tool that can enhance the control of matter at the nanoscale.

Dielectrophoretic characterization of peroxidized retinal pigment epithelial cells as a model of age-related macular degeneration.

BACKGROUND: Age-related macular degeneration (AMD) is a prevalent ocular pathology affecting mostly the elderly population. AMD is characterized by a progressive retinal pigment epithelial (RPE) cell degeneration, mainly caused by an impaired antioxidative defense. One of the AMD therapeutic procedures involves injecting healthy RPE cells into the subretinal space, necessitating pure, healthy RPE cell suspensions. This study aims to electrically characterize RPE cells to demonstrate a possibility using simulations to separate healthy RPE cells from a mixture of healthy/oxidized cells by dielectrophoresis. METHODS: BPEI-1 rat RPE cells were exposed to hydrogen peroxide to create an in-vitro AMD cellular model. Cell viability was evaluated using various methods, including microscopic imaging, impedance-based real-time cell analysis, and the MTS assay. Healthy and oxidized cells were characterized by recording their dielectrophoretic spectra, and electric cell parameters (crossover frequency, membrane conductivity and permittivity, and cytoplasm conductivity) were computed. A COMSOL simulation was performed on a theoretical microfluidic-based dielectrophoretic separation chip using these parameters. RESULTS: Increasing the hydrogen peroxide concentration shifted the first crossover frequency toward lower values, and the cell membrane permittivity progressively increased. These changes were attributed to progressive membrane peroxidation, as they were diminished when measured on cells treated with the antioxidant N-acetylcysteine. The changes in the crossover frequency were sufficient for the efficient separation of healthy cells, as demonstrated by simulations. CONCLUSIONS: The study demonstrates that dielectrophoresis can be used to separate healthy RPE cells from oxidized ones based on their electrical properties. This method could be a viable approach for obtaining pure, healthy RPE cell suspensions for AMD therapeutic procedures.

Electric-Field-Driven Localization of Molecular Nanowires in Wafer-Scale Nanogap Electrodes.

As integrated circuits continue to scale toward the atomic limit, bottom-up processes, such as epitaxial growth, have come to feature prominently in their fabrication. At the same time, chemistry has developed highly tunable molecular semiconductors that can perform the functions of ultimately scaled circuit components. Hybrid techniques that integrate programmable structures comprising molecular components into devices however are sorely lacking. Here we demonstrate a wafer-scale process that directs the localization of a conductive polymer, Mw = 20 kg mol-1 polyaniline, from dilute solutions into 50 nm vertical nanogap device architectures using electric-field-driven self-assembly. The resulting metal-polymer-metal junctions were characterized by electron microscopy, Raman spectroscopy and transport measurements demonstrating that our technique is highly selective, assembling conductive polymers only in electrically activated nanogaps. Our results represent a step toward scalable hybrid nanoelectronics that seamlessly integrate established lithographic top-down fabrication with bottom-up synthesized molecular functional circuit components.

Label-free detection of ConA-induced T-lymphocyte activation at single-cell level by microfluidics.

Lymphocyte activation is critical in regulating immune responses. The resulting T-cell proliferation has been implicated in the pathogenesis of a variety of autoimmune diseases, such as SLE and rheumatoid arthritis. ConA (concanavalin A)-induced activation has been widely used in the T lymphocytes model of immune-mediated liver injury, autoimmune hepatitis, and so on. In those works, it usually requires fluorescent labeling or cell staining to confirm whether the cells are transformed successfully after medicine treatment to figure out efficacy/pharmacology. The detection preparation steps are time-consuming and have limitations for further proteomic/genomic identifications. Here, a label-free microfluidic method is established to detect lymphocyte activation degree. The lymphocyte and ConA-activated lymphocyte were investigated by a microfluidic device. According to where single cells in the sample were captured in the designed channel, lymphocyte and ConA-activated samples are differentiated and characterized by population electric field factors, 2.08 × 104 and 2.21 × 104 V/m, respectively. Furthermore, salidroside, a herbal medicine that was documented to promote the transformation, was used to treat lymphocyte cells, and the treated cell population is detected to be 2.67 × 104 V/m. The characterization indicates an increasing trend with the activation degree. The result maintains a high consistency with traditional staining methods with transformed cells of 15.8%, 28.8%, and 48.3% in each cell population. Dielectrophoresis is promising to work as a tool for detecting lymphocyte transformation and medical efficacy detection.

Oligo cyc-DEP: On-chip cyclic immunofluorescence profiling of cell-derived nanoparticles.

We present a follow-on technique for the cyclic-immunofluorescence profiling of suspension particles isolated using dielectrophoresis. The original lab-on-chip technique ("cyc-DEP" [cyclic immunofluorescent imaging on dielectrophoretic chip]) was designed for the multiplex surveillance of circulating biomarkers. Nanoparticles were collected from low-volume liquid biopsies using microfluidic dielectrophoretic chip technology. Subsequent rounds of cyclic immunofluorescent labeling and quenching were imaged and quantified with a custom algorithm to detect multiple proteins. While cyc-DEP improved assay multiplicity, long runtimes threatened its clinical adoption. Here, we modify the original cyc-DEP platform to reduce assay runtimes. Nanoparticles were formulated from human prostate adenocarcinoma cells and collected using dielectrophoresis. Three proteins were labeled on-chip with a mixture of short oligonucleotide-conjugated antibodies. The sample was then incubated with complementary fluorophore-conjugated oligonucleotides, which were dehybridized using an ethylene carbonate buffer after each round of imaging. Oligonucleotide removal exhibited an average quenching efficiency of 98 ± 3% (n = 12 quenching events), matching the original cyc-DEP platform. The presented "oligo cyc-DEP" platform achieved clinically relevant sample-to-answer times, reducing the duration for three rounds of cyclic immunolabeling from approximately 20 to 6.5 h-a 67% decrease attributed to rapid fluorophore removal and the consolidated co-incubation of antibodies.

A stepwise multi-stage continuous dielectrophoresis separation microfluidic chip with microfilter structures.

The separation of target microparticles using microfluidic systems owns extensive applications in biomedical, chemical, and materials science fields. Integration of microfluidic sorting systems employing dielectrophoresis (DEP) technology has been widely investigated. However, enhancing separation efficiency, purity, stability, and integration remains a pressing issue. This study proposes a stepwise multi-stage continuous DEP separation microfluidic chip with a microfilter structure. By leveraging a stepwise electrode configuration, a gradient electric field is generated to drive target microparticles along the electric field gradient, thereby enhancing separation efficiency. Innovative integration of a microfilter structure facilitates simultaneous filtration and improves flow field distribution, thus enhancing system stability. Through the synergistic effect of stepwise electrodes and the microfilter structure, superior coupling of electric and flow fields is achieved, consequently improving the sorting purity, separation efficiency, and system stability of the DEP-based microfluidic sorting system. Validation through simulation and separation of polystyrene microspheres demonstrates the excellent particle separation performance of the proposed system. It evidently shows potential for seamless extension to various biological microparticle sorting applications, harboring significant prospects in the biomedical domain field.

Electrochemical Impedance Spectroscopy in the Determination of the Dielectric Properties of Tau-441 Protein for Dielectrophoresis Response Prediction.

This study employs electrochemical impedance spectroscopy (EIS) to probe the behavior of Tau-441 protein, a key component implicated in Alzheimer's disease. Through meticulous experimentation and analysis, the impedance of Tau-441 protein suspension revealed a conductivity peak value of 1.02 S/m. The study demonstrates a high level of specificity and selectivity, particularly within the challenging nanomolar concentration range. Additionally, the EIS method enabled the prediction of Tau-441 protein's dielectrophoresis (DEP) response and the determination of the associated frequency range of 1 kHz to 1 MHz. These findings contribute to advancing our understanding of the molecular intricacies surrounding Tau-441 and hold promise for unraveling implications related to Alzheimer's disease. This study establishes a robust foundation for future research on neurodegenerative disease and biosciences, offering valuable insights into the electrochemical dynamics of Tau-441 protein.

Dielectrophoretic separation and purification: From colloid and biological particles to droplets.

It is indispensable to realize the high level of purification and separation, so that objective particles, such as malignant cells, harmful bacteria, and special proteins or biological molecules, could satisfy the high precise measurement in the pharmaceutical analysis, clinical diagnosis, targeted therapy, and food defense. In addition, this could reveal the intrinsic nature and evolution mechanisms of individual biological variations. Consequently, many techniques related to optical tweezers, microfluidics, acoustophoresis, and electrokinetics can be broadly used to achieve micro- and nano-scale particle separations. Dielectrophoresis (DEP) has been used for various manipulation, concentration, transport, and separation processes of biological particles owing to its early development, mature theory, low cost, and high throughput. Although numerous reviews have discussed the biological applications of DEP techniques, comprehensive descriptions of micro- and nano-scale particle separations feature less frequently in the literature. Therefore, this review summarizes the current state of particle separation attention to relevant technological developments and innovation, including theoretical simulation, microchannel structure, electrode material, pattern and its layout. Moreover, a brief overview of separation applications using DEP in combination with other technologies is also provided. Finally, conclusions, future guidelines, and suggestions for potential promotion are highlighted.

Collection of serum albumin aggregate nanoparticles from human plasma by dielectrophoresis.

Dielectrophoresis (DEP) is a fast and reliable nanoparticle recovery method that utilizes nonuniform electric fields to manipulate particles based on their material composition and size, enabling recovery of biologically-derived nanoparticles from plasma for diagnostic applications. When applying DEP to undiluted human plasma, collection of endogenous albumin proteins was observed at electric field gradients much lower than predicted by theory to collect molecular proteins. To understand this collection, nanoparticle tracking analysis of bovine serum albumin (BSA) dissolved in 0.5× phosphate-buffered saline was performed and showed that albumin spontaneously formed aggregate nanoparticles with a mean diameter of 237 nm. These aggregates experienced a dielectrophoretic force as a function of aggregate radius rather than the diameter of individual protein molecules which contributed to their collection. In high conductance buffer (6.8 mS/cm), DEP was able to move these aggregates into regions of high electric field gradient, and in lower conductance buffer (0.68 mS/cm), these aggregates could be moved into high or low gradient regions depending on the applied frequency. Disruption of BSA aggregates using a nonionic detergent significantly decreased the particle diameter, resulting in decreased dielectrophoretic collection of albumin which increased the collection consistency of particles of interest. These results provide techniques to manipulate albumin aggregates via DEP, which impacts collection of diagnostic biomarkers.

Measurement and analysis of ionic leakage profiles in refrigerated human red blood cells using dielectrophoresis and inductively coupled mass spectroscopy.

Human red blood cells (RBCs) undergo ionic leakage through passive diffusion during refrigerated storage, affecting their quality and health. We investigated the dynamics of ionic leakage in human RBCs over a 20-day refrigerated storage period using extracellular ion quantification and dielectrophoresis (DEP). Four type O- human blood donors were examined to assess the relationship between extracellular ion concentrations (Na+, K+, Mg2+, Ca2+, and Fe2+), RBC cytoplasm conductivity, and membrane conductance. A consistent negative correlation between RBC cytoplasm conductivity and membrane conductance, termed the "ionic leakage profile" (ILP), was observed across the 20-day storage period. Specifically, we noted a gradual decline in DEP-measured RBC cytoplasm conductivity alongside an increase in membrane conductance. Further examination of the electrical origins of this ILP using inductively coupled plasma mass spectrometry revealed a relative decrease in extracellular Na+ concentration and an increase in K+ concentration over the storage period. Correlation of these extracellular ion concentrations with DEP-measured RBC electrical properties demonstrated a direct link between changes in the cytoplasmic and membrane domains and the leakage and transport of K+ and Na+ ions across the cell membrane. Our analysis suggests that the inverse correlation between RBC cytoplasm and membrane conductance is primarily driven by the passive diffusion of K+ from the cytoplasm and the concurrent diffusion of Na+ from the extracellular buffer into the membrane, resulting in a conductive reduction in the cytoplasmic domain and a subsequent increase in the membrane. The ILP's consistent negative trend across all donors suggests that it could serve as a metric for quantifying blood bank storage age, predicting the quality and health of refrigerated RBCs.

Combination of an Optically Induced Dielectrophoresis (ODEP) Mechanism and a Laminar Flow Pattern in a Microfluidic System for the Continuous Size-Based Sorting and Separation of Microparticles.

Optically induced dielectrophoresis (ODEP)-based microparticle sorting and separation is regarded as promising. However, current methods normally lack the downstream process for the transportation and collection of separated microparticles, which could limit its applications. To address this issue, an ODEP microfluidic chip encompassing three microchannels that join only at the central part of the microchannels (i.e., the working zone) was designed. During operation, three laminar flows were generated in the zone, where two dynamic light bar arrays were designed to sort and separate PS (polystyrene) microbeads of different sizes in a continuous manner. The separated PS microbeads were then continuously transported in laminar flows in a partition manner for the final collection. The results revealed that the method was capable of sorting and separating PS microbeads in a high-purity manner (e.g., the microbead purity values were 89.9 ± 3.7, 88.0 ± 2.5, and 92.8 ± 6.5% for the 5.8, 10.8, and 15.8 µm microbeads harvested, respectively). Overall, this study demonstrated the use of laminar flow and ODEP to achieve size-based sorting, separation, and collection of microparticles in a continuous and high-performance manner. Apart from the demonstration, this method can also be utilized for size-based sorting and the separation of other biological or nonbiological microparticles.

Proposal and performance evaluation of a new parallel plate continuous cell separation device using dielectrophoresis.

Along with the rapid development of cellular biological research in recent years, there has been an urgent need for a high-speed, high-precision method of separating target cells from a highly heterogeneous cell population. Among the various cell separation technologies proposed so far, dielectrophoresis (DEP)-based approaches have shown particular promise because they are noninvasive to cells. We have developed a new DEP-based device to separate large numbers of live and dead cells of the human mammary cell line MCF10A. In this study, we validated the separation performance of this device. The results showed the successful separation of a higher percentage of cells than in previous studies, with a separation efficiency higher than 90%. In the past, there have been no confirmed cases in which a separation rate of over 90% and high-speed processing of a large number of cells were simultaneously achieved. It was shown that the proposed device can process large numbers of cells at high speed and with high accuracy.

Rapid and Sensitive Detection by Combining Electric Field Effects and Surface Plasmon Resonance: A Theoretical Study.

Surface plasmon resonance (SPR) has been extensively employed in biological sensing, environmental detection, as well as chemical industry. Nevertheless, the performance possessed by conventional surface plasmon resonance (SPR) biosensors can be further limited by the transport of analyte molecules to the sensing surface, noteworthily when small molecules or low levels of substances are being detected. In this study, a rapid and highly sensitive SPR biosensor is introduced to enhance the ability of the target analytes' collection by integrating AC electroosmosis (ACEO) and dielectrophoresis (DEP). Both the above-mentioned phenomena principally arise from the generation of the AC electric fields. This generation can be tailored by shaping the interdigitated electrodes (IDEs) that also serve as the SPR biomarker sensing area. The effects exerted by different parameters (e.g., the frequency and voltage of the AC electric field as well as microelectrode structures) are considered in the iSPR (interdigitated SPR) biosensor operation, and the iSPR biosensors are optimized with the sensitivity. The results of this study confirm that the iSPR can efficiently concentrate small molecules into the SPR sensing area, such that SPR reactions achieve an order of magnitude increase, and the detection time is shortened. The rapid and sensitive sensor takes on critical significance in the development of on-site diagnostics in a wide variety of human and animal health applications.

Microfluidic Electroporation Arrays for Investigating Electroporation-Induced Cellular Rupture Dynamics.

Electroporation is pivotal in bioelectrochemistry for cellular manipulation, with prominent applications in drug delivery and cell membrane studies. A comprehensive understanding of pore generation requires an in-depth analysis of the critical pore size and the corresponding energy barrier at the onset of cell rupture. However, many studies have been limited to basic models such as artificial membranes or theoretical simulations. Challenging this paradigm, our study pioneers using a microfluidic electroporation chip array. This tool subjects live breast cancer cell species to a diverse spectrum of alternating current electric field conditions, driving electroporation-induced cell rupture. We conclusively determined the rupture voltages across varying applied voltage loading rates, enabling an unprecedented characterization of electric cell rupture dynamics encompassing critical pore radius and energy barrier. Further bolstering our investigation, we probed cells subjected to cholesterol depletion via methyl-ß-cyclodextrin and revealed a strong correlation with electroporation. This work not only elucidates the dynamics of electric rupture in live cell membranes but also sets a robust foundation for future explorations into the mechanisms and energetics of live cell electroporation.

Electrical signature of heterogeneous human mesenchymal stem cells.

Human mesenchymal stem cells (hMSCs) have gained traction in transplantation therapy due to their immunomodulatory, paracrine, immune-evasive, and multipotent differentiation potential. The inherent heterogeneity of hMSCs poses a challenge for therapeutic treatments and necessitates the identification of robust biomarkers to ensure reproducibility in both in vivo and in vitro experiments. In this study, we utilized dielectrophoresis (DEP), a label-free electrokinetic phenomenon, to investigate the heterogeneity of hMSCs derived from bone marrow (BM) and adipose tissue (AD). The electrical properties of BM-hMSCs were compared to homogeneous mouse fibroblasts (NIH-3T3), human fibroblasts (WS1), and human embryonic kidney cells (HEK-293). The DEP profile of BM-hMSCs differed most from HEK-293 cells. We compared the DEP profiles of BM-hMSCs and AD-hMSCs and found that they have similar membrane capacitances, differing cytoplasm conductivity, and transient slopes. Inducing both populations to differentiate into adipocyte and osteoblast cells revealed that they behave differently in response to differentiation-inducing cytokines. Histology and reverse transcription-quantitative polymerase chain reaction (RT-qPCR) analyses of the differentiation-related genes revealed differences in heterogeneity between BM-hMSCs and AD-hMSCs. The differentiation profiles correlate well with the DEP profiles developed and indicate differences in the heterogeneity of BM-hMSCs and AD-hMSCs. Our results demonstrate that using DEP, membrane capacitance, cytoplasm conductivity, and transient slope can uniquely characterize the inherent heterogeneity of hMSCs to guide robust and reproducible stem cell transplantation therapies.

Dielectrophoresis: Measurement technologies and auxiliary sensing applications.

Dielectrophoresis (DEP), which arises from the interaction between dielectric particles and an aqueous solution in a nonuniform electric field, contributes to the manipulation of nano and microparticles in many fields, including colloid physics, analytical chemistry, molecular biology, clinical medicine, and pharmaceutics. The measurement of the DEP force could provide a more complete solution for verifying current classical DEP theories. This review reports various imaging, fluidic, optical, and mechanical approaches for measuring the DEP forces at different amplitudes and frequencies. The integration of DEP technology into sensors enables fast response, high sensitivity, precise discrimination, and label-free detection of proteins, bacteria, colloidal particles, and cells. Therefore, this review provides an in-depth overview of DEP-based fabrication and measurements. Depending on the measurement requirements, DEP manipulation can be classified into assistance and integration approaches to improve sensor performance. To this end, an overview is dedicated to developing the concept of trapping-on-sensing, improving its structure and performance, and realizing fully DEP-assisted lab-on-a-chip systems.

Dielectrophoretic separation/classification/focusing of microparticles using electrified lab-on-a-disc platforms.

BACKGROUND: Separation, classification, and focusing of microparticles are essential issues in microfluidic devices that can be implemented in two categories: using labeling and label-free methods. Label-free methods differentiate microparticles based on their inherent properties, including size, density, shape, electrical conductivity/permittivity, and magnetic susceptibility. Dielectrophoresis is an advantageous label-free technique for this objective. Besides, centrifugal microfluidic devices exploit centrifugal forces to move liquid and particles. The simultaneous combination of dielectrophoretic and centrifugal forces exerted on microparticles still needs to be scrutinized more to predict their trajectories in such devices. RESULTS: An integrated system utilizing two categories in microfluidics is proposed: dielectrophoretic manipulation of microparticles and centrifugal-driven microfluidics, followed by a numerical analysis. In this regard, we assumed a rectangular microchannel with internal unilateral planar electrodes equipped with three equal-sized outlets placed radially on a centrifugal platform where microparticles flow toward the disc's outer edge. The effect of different coordinate-based parameters, including radial and lateral distances (X and Y offsets)/tilting angles toward the radius direction (α), on the particles' movement was investigated. Additionally, the effect of operational parameters, including applied voltage, the microchannel width, the number of enabled electrodes, the diameter of particles, and the configuration of electrodes, were analyzed, and the distributions of particles toward the outlets were monitored. It was found that enhanced particle focusing becomes possible at lower rotation speeds and higher electric field values. Furthermore, the proposed centrifugal-DEP system's efficiency for classifying red blood cells/platelets and Live/Dead yeast cells systems was evaluated. SIGNIFICANCE: Our integrated system is introduced as a novel method for focusing and classifying various microparticles with no need for sheath flows, having the ability to conduct particles at desired routes and focusing width. Furthermore, the system effectively separates various bioparticles and offers ease of operation and high-efficiency throughput over conventional dielectrophoretic devices.

Small extracellular vesicles detection using dielectrophoresis-based microfluidic chip for diagnosis of breast cancer.

Small extracellular vesicles (sEVs) reflect the genotype and phenotype of original cells and are biomarkers for early diagnosis and treatment monitoring of tumors. Yet, their small size and low density make them difficult to isolate and detect in body fluid samples. This study proposes a novel acDEP-Exo chip filled with transparent micro-beads, which formed a non-uniform electrical field, and finally achieved rapid, sensitive, and tunable sEVs capture and detection. The method requires only 20-50 µL of sample, achieved a limit of detection (LOD) of 161 particles/µL, and can detect biomarkers within 13 min. We applied the chip to analyze the two markers of sEV's EpCAM and MUC1 in clinical plasma samples from breast cancer (BC) patients and healthy volunteers and found that the combined evaluation of sEV's biomarkers has extremely high sensitivity, specificity and accuracy. The present study introduces an alternative approach to sEVs isolation and detection, has a great potential in real-time sEVs-based liquid biopsy.

Cell Electrokinetic Fingerprint: A Novel Approach Based on Optically Induced Dielectrophoresis (ODEP) for In-Flow Identification of Single Cells.

A novel optically induced dielectrophoresis (ODEP) system that can operate under flow conditions is designed for automatic trapping of cells and subsequent induction of 2D multi-frequency cell trajectories. Like in a "ping-pong" match, two virtual electrode barriers operate in an alternate mode with varying frequencies of the input voltage. The so-derived cell motions are characterized via time-lapse microscopy, cell tracking, and state-of-the-art machine learning algorithms, like the wavelet scattering transform (WST). As a cell-electrokinetic fingerprint, the dynamic of variation of the cell displacements happening, over time, is quantified in response to different frequency values of the induced electric field. When tested on two biological scenarios in the cancer domain, the proposed approach discriminates cellular dielectric phenotypes obtained, respectively, at different early phases of drug-induced apoptosis in prostate cancer (PC3) cells and for differential expression of the lectine-like oxidized low-density lipoprotein receptor-1 (LOX-1) transcript levels in human colorectal adenocarcinoma (DLD-1) cells. The results demonstrate increased discrimination of the proposed system and pose an additional basis for making ODEP-based assays addressing cancer heterogeneity for precision medicine and pharmacological research.

Microfluidics identify moso bamboo and henon bamboo by leaf protoplast subpopulations with single-cell analysis.

Current techniques identifying herbal medicine species require marker labeling or lack systematical accuracy (expert authentication). There is an emerging interest in developing an accurate and label-free tool for herbal medicine authentication. Here, a high-resolution microfluidic-based method is developed for identifying herbal species by protoplast subpopulations. Moso bamboo and henon bamboo are used as a model to be differentiated based on protoplast. Their biophysical properties factors are characterized to be 7.09 (± 0.39) × 108 V/m2 and 6.54 (± 0.26) × 108 V/m2, respectively. Their biophysical distributions could be distinguished by the Cramér-von Mises criterion with a 94.60% confidence level. The subpopulations of each were compared with conventional flow cytometry indicating the existence of subpopulations and the differences between the two species. The subsets divided by a biophysical factor of 8.05(± 0.51) × 108 V/m2 suggest good consistency with flow cytometry. The work demonstrated the possibility of microfluidics manipulation on protoplast for medication safety use taking advantage of dielectrophoresis. The device is promising in developing a reliable and accurate way of identifying herbal species with difficulties in authentication.