Nano-constriction device for rapid protein preconcentration in physiological media through a balance of electrokinetic forces

We describe a methodology to steeply enhance streptavidin protein preconcentration within physiological media over that achieved by negative dielectrophoresis (NDEP) through utilizing a DC offset to the AC field at nanoscale constriction gap devices. Within devices containing approximately 50-nm constriction gaps, we find that the addition of a critical DC field offset (1.5 V/cm) to the NDEP condition (～200 V(pp) /cm at 1 MHz) results in an exponentially enhanced extent of protein depletion across the device to cause a rapid and steeply rising degree of protein preconcentration. Under these conditions, an elliptical-shaped protein depletion zone that is extended along the device centerline axis forms instantaneously around the constrictions to result in protein preconcentration along the constriction sidewall direction. Through a potential energy diagram to describe the electrokinetic force balance across the device, we find that the potential energy barrier due to NDEP is gradually tilted upon addition of DC fields, to cause successively steeper potential wells along the sidewall direction for devices containing smaller constriction gaps. Hence, for approximately 50-nm constriction gaps at a critical DC field, the ensuing narrow and deep potential energy wells enable steep protein preconcentration, due to depletion over an exponentially enhanced extent across the device.     

High-resolution analyses of cell fusion dynamics in a biochip

In this study, we analyzed the electrofusion of two cells in a biochip that has been developed to perform the capture by dielectrophoresis and the electrofusion of pairs of cells. The good transparency of the microsystem allowed analyzing the details of the fusion events. By staining one of the cells, the mixing of the two cytosols could be observed during the electrofusion experiment. We show for the first time the rapidity of the mixing of the two cytosols: less than 5 s under our experimental conditions. By comparing these experimental results to a numerical simulation, we found that the rate of this phenomenon is compatible with a diffusion-only mechanism, showing that during the fusion, the two cell membranes in contact are affected by very rapid structural changes and do not limit the exchange of the cytosols between the two cells. A point of interest is the use of dielectric structures to concentrate the electric field and of positive dielectrophoresis to capture cells in the area where the electric field is more intense. This technique allows the increase of the cell-to-cell contact and limits cell cytosol leakages during the fusion process.     

Dielectrophoretic isolation of DNA and nanoparticles from blood

The ability to effectively detect disease-related DNA biomarkers and drug delivery nanoparticles directly in blood is a major challenge for viable diagnostics and therapy monitoring. A DEP method has been developed which allows the rapid isolation, concentration and detection of DNA and nanoparticles directly from human and rat whole blood. Using a microarray device operating at 20 V peak-to-peak and 10 kHz, a wide range of high molecular weight (HMW)-DNA and nanoparticles were concentrated into high-field regions by positive DEP, while the blood cells were concentrated into the low-field regions by negative DEP. A simple fluidic wash removes the blood cells while the DNA and nanoparticles remain concentrated in the DEP high-field regions where they can be detected by fluorescence. HMW-DNA could be detected at 260 ng/mL, which is a detection level suitable for analysis of disease-related cell-free circulating DNA biomarkers. Fluorescent 40 nm nanoparticles could be detected at 9.5 × 10(9) particles/mL, which is a level suitable for monitoring drug delivery nanoparticles. The ability to rapidly isolate and detect DNA biomarkers and nanoparticles from undiluted whole blood will benefit many diagnostic applications by significantly reducing sample preparation time and complexity.     

Microfluidic device embedding electrodes for dielectrophoretic manipulation of cells‐A review

Microfluidic device embedding electrodes realizes cell manipulation with the help of dielectrophoresis. Cell manipulation is an important technology for cell sorting and cell population purification. Till now, the theory of dielectrophoresis has been greatly developed. Microfluidic devices with various arrangements of electrodes have been reported from the beginning of the single non‐uniform electric field to the later multiple physical fields. This paper reviews the research status of microfluidic device embedding electrodes for cell manipulation based on dielectrophoresis. Firstly, the working principle of dielectrophoresis is explained. Next, cell manipulation approaches based on dielectrophoresis are introduced. Then, different types of electrode arrangements in the microfluidic device for cell manipulation are discussed, including planar, multilayered and microarray dot electrodes. Finally, the future development trend of the dielectrophoresis with the help of microfluidic devices is prospected. With the rapid development of microfluidic technology, in the near future, high precision, high throughput, high efficiency, multifunctional, portable, economical and practical microfluidic dielectrophoresis will be widely used in the fields of biology, medicine, agriculture and so on.     

On the design of deterministic dielectrophoresis for continuous separation of circulating tumor cells from peripheral blood cells

Detection and analysis of circulating tumor cells (CTCs) have emerged as a promising way to diagnose cancer, study its cellular mechanism, and test or develop potential treatments. However, the rarity of CTCs among peripheral blood cells is a big challenge toward CTC detection. In addition, in cases where there is similar size range between certain types of CTCs (e.g. breast cancer cells) and white blood cells (WBCs), high‐resolution techniques are needed. In the present work, we propose a deterministic dielectrophoresis (DEP) method that combines the concept of deterministic lateral displacement (DLD) and insulator‐based dielectrophoresis (iDEP) techniques that rely on physical markers such as size and dielectric properties to differentiate different type of cells. The proposed deterministic DEP technology takes advantage of frequency‐controlled AC electric field for continuous separation of CTCs from peripheral blood cells. Utilizing numerical modeling, different aspects of coupled DLD‐DEP design such as the required applied voltages, velocities, and geometrical parameters of DLD arrays of microposts are investigated. Regarding the inevitable difference and uncertainty ranges for the reported crossover frequencies of cells, a comprehensive analysis is conducted on applied electric field frequency as design's determinant factor. Deterministic DEP design provides continuous sorting of CTCs from WBCs even with similar size and has the future potential for high throughput and efficiency.     

On the use of correction factors for the mathematical modeling of insulator based dielectrophoretic devices

Mathematical modeling is a fundamental component in the development of new microfluidics techniques and devices. Modeling allows for the rapid testing of new system configurations while saving resources. Microscale electrokinetic (EK) techniques have significantly benefited by the advances in modeling programs and software packages. However, EK phenomena are complex to model, as they dynamically affect system characteristics, including the physical properties of the particles and fluid within the system. Insulator‐based dielectrophoresis (iDEP) is an EK technique that has received important attention during the last two decades. In particular, numerous research groups that study iDEP systems employ a combination of modeling and experimentation for developing new iDEP systems. An important fraction of these research groups has adopted the practice of employing “correction factors” to account for EK phenomena that cannot be accurately predicted in their models due to model complexity and limitations in computing resources. The present review article aims to provide the reader with an overview of the most common approaches in the use of correction factors for the modeling of iDEP systems.     

AC dielectrophoretic deformable particle-particle interactions and their relative motions

This paper develops a numerical simulation model to research the deformable particle-particle interactions caused by dielectrophoresis (DEP) under AC electric fields. The DEP force is calculated by using Maxwell stress tensor method, and the hydrodynamic force is obtained by calculating the hydrodynamic stress tensor. Simulation results show that the DEP interactive motion will facilitate the particles forming particle chains that are parallel to the electric field, and the particles with low shear modulus present a lower x-component velocity. Also, the electric field intensity and particles radius have some effects on the DEP motions, and for different particles, smaller particles with larger electric field intensity easily reach a larger velocity. The numerical research may provide universal guidance for biological cells manipulation and assembly.     

Fast immobilization of probe beads by dielectrophoresis-controlled adhesion in a versatile microfluidic platform for affinity assay

The use of probe beads for lab-on-chip affinity assays is very interesting from a practical point of view. It is easier to handle and trap beads than molecules in microfluidic systems. We present a method for the immobilization of probe beads at defined areas on a chip using dielectrophoresis (DEP)-controlled adhesion. The method is fast, i.e., it takes between 10 and 120 s--depending on the protocol--to functionalize a chip surface at defined areas. The method is versatile, i.e., it works for beads with different types of probe molecule coatings. The immobilization is irreversible, i.e., the retained beads are able to withstand high flow velocities in a flow-through device even after the DEP voltage is turned off, thus allowing the use of conventional high-conductivity analyte buffers in the following assay procedure. We demonstrate the on-chip immobilization of fluorescent beads coated with biotin, protein A, and goat-antimouse immunoglobulin G (IgG). The number of immobilized beads at an electrode array can be determined from their fluorescence signal. Further, we use this method to demonstrate the detection of streptavidin and mouse IgG. Finally, we demonstrate the feasibility of the parallel detection of different analyte molecules on the same chip.     

Gravitation-driven stress-reduced cell handling

We present a simple lab-on-chip device for handling small samples of delicate cells, e.g. stem cells. It uses a combination of sedimentation and dielectrophoresis. The transport of cells is driven by gravitation. Dielectrophoresis uses radio-frequency electric fields for generating particle-selective forces dependent on size and polarisability. Electrodes along the channels hold particles and/or cells in a defined position and deflect them towards different outlets. The absence of external pumping and the integration of injection and sampling ports allow the processing of tiny sample volumes. Various functions are demonstrated, such as contact-free cell trapping and cell/particle sorting. Pairs of human cells and antibody-coated beads, as they are formed for T cell activation, are separated from unbound beads. The cells experience only low stress levels compared with the stress levels in dielectrophoresis systems, where transport depends on external pumping. Our device is a versatile yet simple tool that finds applications in cellular biotechnology, in particular when an economic solution is required. Figure A simple gravitation-driven lab-on-chip device for the separation of mixed populations of microparticles or cells by negative dielectrophoresis.     

Controlling the mechanics and nanotopography of biocompatible scaffolds through dielectrophoresis with carbon nanotubes

Creating a biocompatible carbon-nanotube polymer scaffold is an area that has a number of potential applications. Herein, a dielectrophoretic approach was pursued to integrate carbon nanotubes into a polymeric material for fabricating a nanoscale composite scaffold with increased and controllable mechanical strength. The adhesion force, which combines the surface energy of the sample and the interfacial energy between the tip and sample, was estimated to be 55.39 +/- 6.72 nN away from the center of the protrusions at a distance of 0.5 microm while being 24.01 +/- 4.45 nN at the center. The adhesion force for the center of the cavities was 42.47 +/- 6.91 and 88.21 +/- 15.05 nN at 0.5 microm away from the center. NIH 3T3 fibroblast cells were then utilized to test the cellular biocompatibility of this multiwalled carbon nanotubes (MWCNTs) film. Cells were cultured on the surface and then their attachment, spreading, and proliferation behaviors were observed. This nanotube-polymer scaffolding approach has a wide range of potential applications including in complex device fabrication as well as in developing biomimetic and tissue engineering scaffolds, and artificial organs.