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1.
Integrating different steps on a chip for cell manipulations and sample preparation is of foremost importance to fully take advantage of microfluidic possibilities, and therefore make tests faster, cheaper and more accurate. We demonstrated particle manipulation in an integrated microfluidic device by applying hydrodynamic, electroosmotic (EO), electrophoretic (EP), and dielectrophoretic (DEP) forces. The process involves generation of fluid flow by pressure difference, particle trapping by DEP force, and particle redirect by EO and EP forces. Both DC and AC signals were applied, taking advantages of DC EP, EO and AC DEP for on-chip particle manipulation. Since different types of particles respond differently to these signals, variations of DC and AC signals are capable to handle complex and highly variable colloidal and biological samples. The proposed technique can operate in a high-throughput manner with thirteen independent channels in radial directions for enrichment and separation in microfluidic chip. We evaluated our approach by collecting Polystyrene particles, yeast cells, and E. coli bacteria, which respond differently to electric field gradient. Live and dead yeast cells were separated successfully, validating the capability of our device to separate highly similar cells. Our results showed that this technique could achieve fast pre-concentration of colloidal particles and cells and separation of cells depending on their vitality. Hydrodynamic, DC electrophoretic and DC electroosmotic forces were used together instead of syringe pump to achieve sufficient fluid flow and particle mobility for particle trapping and sorting. By eliminating bulky mechanical pumps, this new technique has wide applications for in situ detection and analysis.  相似文献   

2.
Optoelectrofluidic field separation (OEFS) of particles under light -intensity gradient (LIG) is reported, where the LIG illumination on the photoconductive layer converts the short-ranged dielectrophoresis (DEP) force to the long-ranged one. The long-ranged DEP force can compete with the hydrodynamic force by alternating current electro-osmosis (ACEO) over the entire illumination area for realizing effective field separation of particles. In the OEFS system, the codirectional illumination and observation induce the levitation effect, compensating the attenuation of the DEP force under LIG illumination by slightly floating particles from the surface. Results of the field separation and concentration of diverse particle pairs (0.82–16 μm) are well demonstrated, and conditions determining the critical radius and effective particle manipulation are discussed. The OEFS with codirectional LIG strategy could be a promising particle manipulation method in many applications where a rapid manipulation of biological cells and particles over the entire working area are of interest.  相似文献   

3.
This paper presents a field-flow method for separating particle populations in a dielectrophoretic (DEP) chip with asymmetric electrodes under continuous flow. The structure of the DEP device (with one thick electrode that defines the walls of the microfluidic channel and one thin electrode), as well as the fabrication and characterization of the device, was previously described. A characteristic of this structure is that it generates an increased gradient of electric field in the vertical plane that can levitate the particles experiencing negative DEP. The separation method consists of trapping one population to the bottom of the microfluidic channel using positive DEP, while the other population that exhibits negative DEP is levitated and flowed out. Viable and nonviable yeast cells were used for testing of the separation method.  相似文献   

4.
The conventional microfluidic H filter is modified with multi-insulating blocks to achieve a flow-through manipulation and separation of microparticles. The device transports particles by exploiting electro-osmosis and electrophoresis, and manipulates particles by utilizing dielectrophoresis (DEP). Polydimethylsiloxane (PDMS) blocks fabricated in the main channel of the PDMS H filter induce a nonuniform electric field, which exerts a negative DEP force on the particles. The use of multi-insulating blocks not only enhances the DEP force generated, but it also increases the controllability of the motion of the particles, facilitating their manipulation and separation. Experiments were conducted to demonstrate the controlled flow direction of particles by adjusting the applied voltages and the separation of particles by size under two different input conditions, namely (i) a dc electric field mode and (ii) a combined ac and dc field mode. Numerical simulations elucidate the electrokinetic and hydrodynamic forces acting on a particle, with theoretically predicted particle trajectories in good agreement with those observed experimentally. In addition, the flow field was obtained experimentally with fluorescent tracer particles using the microparticle image velocimetry (μ-PIV) technique.  相似文献   

5.
The recent development of microfluidic "lab on a chip" devices requiring sample sizes <100 μL has given rise to the need to concentrate dilute samples and trap analytes, especially for surface-based detection techniques. We demonstrate a particle collection device capable of concentrating micron-sized particles in a predetermined area by combining AC electroosmosis (ACEO) and dielectrophoresis (DEP). The planar asymmetric electrode pattern uses ACEO pumping to induce equal, quadrilateral flow directed towards a stagnant region in the center of the device. A number of system parameters affecting particle collection efficiency were investigated including electrode and gap width, chamber height, applied potential and frequency, and number of repeating electrode pairs and electrode geometry. The robustness of the on-chip collection design was evaluated against varying electrolyte concentrations, particle types, and particle sizes. These devices are amenable to integration with a variety of detection techniques such as optical evanescent waveguide sensing.  相似文献   

6.
Multi-target pathogen detection using heterogeneous medical samples require continuous filtering, sorting, and trapping of debris, bioparticles, and immunocolloids within a diagnostic chip. We present an integrated AC dielectrophoretic (DEP) microfluidic platform based on planar electrodes that form three-dimensional (3D) DEP gates. This platform can continuously perform these tasks with a throughput of 3 μL∕min. Mixtures of latex particles, Escherichia coli Nissle, Lactobacillus, and Candida albicans are sorted and concentrated by these 3D DEP gates. Surface enhanced Raman scattering is used as an on-chip detection method on the concentrated bacteria. A processing rate of 500 bacteria was estimated when 100 μl of a heterogeneous colony of 107 colony forming units ∕ml was processed in a single pass within 30 min.  相似文献   

7.
Micro-orifice based cell fusion assures high-yield fusion without compromising the cell viability. This paper examines feasibility of a dielectrophoresis (DEP) assisted cell trapping method for parallel fusion with a micro-orifice array. The goal is to create viable fusants for studying postfusion cell behavior. We fabricated a microfluidic chip that contained a chamber and partition. The partition divided the chamber into two compartments and it had a number of embedded micro-orifices. The voltage applied to the electrodes located at each compartment generated an electric field distribution concentrating in micro-orifices. Cells introduced into each compartment moved toward the micro-orifice array by manipulation of hydrostatic pressure. DEP assisted trapping was used to keep the cells in micro-orifice and to establish cell to cell contact through orifice. By applying a pulse, cell fusion was initiated to form a neck between cells. The neck passing through the orifice resulted in immobilization of the fused cell pair at micro-orifice. After washing away the unfused cells, the chip was loaded to a microscope with stage top incubator for time lapse imaging of the selected fusants. The viable fusants were successfully generated by fusion of mouse fibroblast cells (L929). Time lapse observation of the fusants showed that fused cell pairs escaping from micro-orifice became one tetraploid cell. The generated tetraploid cells divided into three daughter cells. The fusants generated with a smaller micro-orifice (diameter∼2 μm) were kept immobilized at micro-orifice until cell division phase. After observation of two synchronized cell divisions, the fusant divided into four daughter cells. We conclude that the presented method of cell pairing and fusion is suitable for high-yield generation of viable fusants and furthermore, subsequent study of postfusion phenomena.  相似文献   

8.
We report the development and results of a two-step method for sorting cells and small particles in a microfluidic device. This approach uses a single microfluidic channel that has (1) a microfabricated sieve which efficiently focuses particles into a thin stream, followed by (2) a dielectrophoresis (DEP) section consisting of electrodes along the channel walls for efficient continuous sorting based on dielectric properties of the particles. For our demonstration, the device was constructed of polydimethylsiloxane, bonded to a glass surface, and conductive agarose gel electrodes. Gold traces were used to make electrical connections to the conductive gel. The device had several novel features that aided performance of the sorting. These included a sieving structure that performed continuous displacement of particles into a single stream within the microfluidic channel (improving the performance of downstream DEP, and avoiding the need for additional focusing flow inlets), and DEP electrodes that were the full height of the microfluidic walls (“vertical electrodes”), allowing for improved formation and control of electric field gradients in the microfluidic device. The device was used to sort polymer particles and HeLa cells, demonstrating that this unique combination provides improved capability for continuous DEP sorting of particles in a microfluidic device.  相似文献   

9.
Diffusion of autocrine and paracrine signaling molecules allows cells to communicate in the absence of physical contact. This chemical-based, long-range communication serves crucial roles in tissue function, activation of the immune system, and other physiological functions. Despite its importance, few in vitro methods to study cell-cell signaling through paracrine factors are available today. Here, we report the design and validation of a microfluidic platform that enables (i) soluble molecule-cell and/or (ii) cell-cell paracrine signaling. In the microfluidic platform, multiple cell populations can be introduced into parallel channels. The channels are separated by arrays of posts allowing diffusion of paracrine molecules between cell populations. A computational analysis was performed to aid design of the microfluidic platform. Specifically, it revealed that channel spacing affects both spatial and temporal distribution of signaling molecules, while the initial concentration of the signaling molecule mainly affects the concentration of the signaling molecules excreted by the cells. To validate the microfluidic platform, a model system composed of the signaling molecule lipopolysaccharide, mouse macrophages, and engineered human embryonic kidney cells was introduced into the platform. Upon diffusion from the first channel to the second channel, lipopolysaccharide activates the macrophages which begin to produce TNF-α. The TNF-α diffuses from the second channel to the third channel to stimulate the kidney cells, which express green fluorescent protein (GFP) in response. By increasing the initial lipopolysaccharide concentration an increase in fluorescent response was recorded, demonstrating the ability to quantify intercellular communication between 3D cellular constructs using the microfluidic platform reported here. Overall, these studies provide a detailed analysis on how concentration of the initial signaling molecules, spatiotemporal dynamics, and inter-channel spacing affect intercellular communication.  相似文献   

10.
We report on reversible electroporation of cells in a flow-through microfluidic device, whereby the required electric field is delivered through a set of integrated microcapillaries between a centre stream of cells and side streams of liquid electrolytes. The electrolytes are applied with a sine wave voltage and cells flow by the microcapillary openings encounter a burst of ac field with a duration and strength determined by their average speed and spatial proximity to the microcapillary openings, respectively. Effectiveness of the approach is presented through numerical simulations and empirical results on electroporation efficiency and cell viability against various flow rates (exposure time to the field) as well as frequencies and root-mean-square (rms) intensities of the field. High frequencies (80–400 kHz) and high intensities (e.g., 1.6 kV/cm, rms) are identified with increased electroporation efficiency 61% and viability 86% on average. These results suggest that the device demonstrated here with a simple design and robust operation offers a viable platform for flow-through electroporation.  相似文献   

11.
Enrichment of rare cell populations such as Circulating Tumor Cells (CTCs) is a critical step before performing analysis. This paper presents a polymeric microfluidic device with integrated thick Carbon-PolyDimethylSiloxane composite (C-PDMS) electrodes designed to carry out dielectrophoretic (DEP) trapping of low abundance biological cells. Such conductive composite material presents advantages over metallic structures. Indeed, as it combines properties of both the matrix and doping particles, C-PDMS allows the easy and fast integration of conductive microstructures using a soft-lithography approach while preserving O2 plasma bonding properties of PDMS substrate and avoiding a cumbersome alignment procedure. Here, we first performed numerical simulations to demonstrate the advantage of such thick C-PDMS electrodes over a coplanar electrode configuration. It is well established that dielectrophoretic force (FDEP) decreases quickly as the distance from the electrode surface increases resulting in coplanar configuration to a low trapping efficiency at high flow rate. Here, we showed quantitatively that by using electrodes as thick as a microchannel height, it is possible to extend the DEP force influence in the whole volume of the channel compared to coplanar electrode configuration and maintaining high trapping efficiency while increasing the throughput. This model was then used to numerically optimize a thick C-PDMS electrode configuration in terms of trapping efficiency. Then, optimized microfluidic configurations were fabricated and tested at various flow rates for the trapping of MDA-MB-231 breast cancer cell line. We reached trapping efficiencies of 97% at 20 μl/h and 78.7% at 80 μl/h, for 100 μm thick electrodes. Finally, we applied our device to the separation and localized trapping of CTCs (MDA-MB-231) from a red blood cells sample (concentration ratio of 1:10).  相似文献   

12.
Electrokinetic transport of cylindrical cells under dc electric fields in a straight microfluidic channel is experimentally and numerically investigated with emphasis on the dielectrophoretic (DEP) effect on their orientation variations. A two-dimensional multiphysics model, composed of the Navier–Stokes equations for the fluid flow and the Laplace equation for the electric potential defined in an arbitrary Lagrangian–Eulerian framework, is employed to capture the transient electrokinetic motion of cylindrical cells. The numerical predictions of the particle transport are in quantitative agreement with the obtained experimental results, suggesting that the DEP effect should be taken into account to study the electrokinetic transport of cylindrical particles even in a straight microchannel with uniform cross-sectional area. A comprehensive parametric study indicates that cylindrical particles would experience an oscillatory motion under low electric fields. However, they are aligned with their longest axis parallel to the imposed electric field under high electric fields due to the induced DEP effect.  相似文献   

13.
In this study, a 3D passivated-electrode, insulator-based dielectrophoresis microchip (3D πDEP) is presented. This technology combines the benefits of electrode-based DEP, insulator-based DEP, and three dimensional insulating features with the goal of improving trapping efficiency of biological species at low applied signals and fostering wide frequency range operation of the microfluidic device. The 3D πDEP chips were fabricated by making 3D structures in silicon using reactive ion etching. The reusable electrodes are deposited on second glass substrate and then aligned to the microfluidic channel to capacitively couple the electric signal through a 100 μm glass slide. The 3D insulating structures generate high electric field gradients, which ultimately increases the DEP force. To demonstrate the capabilities of 3D πDEP, Staphylococcus aureus was trapped from water samples under varied electrical environments. Trapping efficiencies of 100% were obtained at flow rates as high as 350 μl/h and 70% at flow rates as high as 750 μl/h. Additionally, for live bacteria samples, 100% trapping was demonstrated over a wide frequency range from 50 to 400 kHz with an amplitude applied signal of 200 Vpp. 20% trapping of bacteria was observed at applied voltages as low as 50 Vpp. We demonstrate selective trapping of live and dead bacteria at frequencies ranging from 30 to 60 kHz at 400 Vpp with over 90% of the live bacteria trapped while most of the dead bacteria escape.  相似文献   

14.
In this paper, we propose a continuous flow droplet-based microfluidic platform for magnetic particle-based assays by employing in-droplet washing. The droplet-based washing was implemented by traversing functionalized magnetic particles across a laterally merged droplet from one side (containing sample and reagent) to the other (containing buffer) by an external magnetic field. Consequently, the magnetic particles were extracted to a parallel-synchronized train of washing buffer droplets, and unbound reagents were left in an original train of sample droplets. To realize the droplet-based washing function, the following four procedures were sequentially carried in a droplet-based microfluidic device: parallel synchronization of two trains of droplets by using a ladder-like channel network; lateral electrocoalescence by an electric field; magnetic particle manipulation by a magnetic field; and asymmetrical splitting of merged droplets. For the stable droplet synchronization and electrocoalescence, we optimized droplet generation conditions by varying the flow rate ratio (or droplet size). Image analysis was carried out to determine the fluorescent intensity of reagents before and after the washing step. As a result, the unbound reagents in sample droplets were significantly removed by more than a factor of 25 in the single washing step, while the magnetic particles were successfully extracted into washing buffer droplets. As a proof-of-principle, we demonstrate a magnetic particle-based immunoassay with streptavidin-coated magnetic particles and fluorescently labelled biotin in the proposed continuous flow droplet-based microfluidic platform.  相似文献   

15.
Alternating current (AC) dielectrophoresis (DEP) experiments for biological particles in microdevices are typically done at a fixed frequency. Reconstructing the DEP response curve from static frequency experiments is laborious, but essential to ascertain differences in dielectric properties of biological particles. Our lab explored the concept of sweeping the frequency as a function of time to rapidly determine the DEP response curve from fewer experiments. For the purpose of determining an ideal sweep rate, homogeneous 6.08 μm polystyrene (PS) beads were used as a model system. Translatability of the sweep rate approach to ∼7 μm red blood cells (RBC) was then verified. An Au/Ti quadrapole electrode microfluidic device was used to separately subject particles and cells to 10Vpp AC electric fields at frequencies ranging from 0.010 to 2.0 MHz over sweep rates from 0.00080 to 0.17 MHz/s. PS beads exhibited negative DEP assembly over the frequencies explored due to Maxwell-Wagner interfacial polarizations. Results demonstrate that frequency sweep rates must be slower than particle polarization timescales to achieve reliable incremental polarizations; sweep rates near 0.00080 MHz/s yielded DEP behaviors very consistent with static frequency DEP responses for both PS beads and RBCs.  相似文献   

16.
Lewpiriyawong N  Yang C 《Biomicrofluidics》2012,6(1):12807-128079
The recent development of microfluidic “lab on a chip” devices requires the need to continuously separate submicron particles. Here, we present a PDMS microfluidic device with sidewall conducting PDMS (AgPDMS) composite electrodes capable of separating submicron particles in hydrodynamic flow. In particular, the device can service dual functions. First, the AgPDMS composite electrodes embedded in a sidewall of the device channel allow for performing AC-dielectrophoretic (DEP) characterization through direct microscopic observation of particle behavior. Characterization experiments are carried out for numerous parameters including particle size, medium conductivity, and AC field frequency to reveal important dielectrophoresis DEP information in terms of the crossover frequency and positive/negative DEP behavior under specific frequencies. Second, the device offers an advantage that sidewall AgPDMS composite electrodes can produce strong DEP effects throughout the entire channel height, and thus the robustness of the on-chip particle separation is demonstrated for continuous separation in a flowing mixture of 0.5 and 5 μm particles with 100% separation efficiency.  相似文献   

17.
Assessment of the microbial safety of water resources is among the most critical issues in global water safety. As the current detection methods have limitations such as high cost and long process time, new detection techniques have transpired among which microfluidics is the most attractive alternative. Here, we show a novel hybrid dielectrophoretic (DEP) system to separate and detect two common waterborne pathogens, Escherichia coli (E. coli), a bacterium, and Cryptosporidium parvum (C. parvum), a protozoan parasite, from water. The hybrid DEP system integrates a chemical surface coating with a microfluidic device containing inter-digitated microelectrodes to impart positive dielectrophoresis for enhanced trapping of the cells. Trimethoxy(3,3,3-trifluoropropyl) silane, (3-aminopropyl)triethoxysilane, and polydiallyl dimethyl ammonium chloride (p-DADMAC) were used as surface coatings. Static cell adhesion tests showed that among these coatings, the p-DADMAC-coated glass surface provided the most effective cell adhesion for both the pathogens. This was attributed to the positively charged p-DADMAC-coated surface interacting electrostatically with the negatively charged cells suspended in water leading to increased cell trapping efficiency. The trapping efficiency of E. coli and C. parvum increased from 29.0% and 61.3% in an uncoated DEP system to 51.9% and 82.2% in the hybrid DEP system, respectively. The hybrid system improved the cell trapping by encouraging the formation of cell pearl-chaining. The increment in trapping efficiency in the hybrid DEP system was achieved at an optimal frequency of 1 MHz and voltage of 2.5 Vpp for C. parvum and 2 Vpp for E. coli, the latter is lower than 2.5 Vpp and 7 Vpp, respectively, utilized for obtaining similar efficiency in an uncoated DEP system.  相似文献   

18.
This work describes the development of a prototypic microfluidic platform for the generation of stepwise concentration gradients of drugs. A sensitive apoptotic analysis method is integrated into this microfluidic system for studying apoptosis of HeLa cells under the influence of anticancer drug, etoposide, with various concentrations in parallel; it measures the yellow fluorescent protein∕cyan fluorescent protein fluorescence resonance energy transfer (FRET) signal that responds to the activation of caspase-3, an indicator of cell apoptosis. Sets of microfluidic valves on the chip generate stepwise concentration gradient of etoposide in various cell-culture microchambers. The FRET signals from multiple chambers are simultaneously monitored under a fluorescent microscope for long-time observation and the on-chip results are compared with those from 96-well plate study and the methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay. The microfluidic platform shows several advantages including high-throughput capacity, low drug consumption, and high sensitivity.  相似文献   

19.
This Special Topic section is a compilation of several original contributions covering both fundamental and practical aspects of electrokinetic microfluidic phenomena that were presented during the Electrokinetics and Microfluidics sessions held at the conference.Electrokinetics is currently the mechanism of choice for the manipulation of fluids as well as colloidal and biological particles at microscale and nanoscale dimensions.1 The popularity of electrokinetics is perhaps not so surprising as electrodes are easy to fabricate and embed into microfluidic chips, thus allowing the entire fluid and particle actuation mechanism to be completely integrated into the device. In addition, driving microfluidics with electric fields is relatively straightforward and allows for precise actuation. Nevertheless, considerable challenges remain in understanding the complex mechanisms associated with the hydrodynamics of conducting and dielectric fluids and particles under the influence of electric fields. Concomitantly, there has been an exponential increase in research and development in this field along both fundamental and applied themes in the past five years.This sustained growth in the microfluidics community of electrokinetics research has led to a sequel to the first Electrokinetic Phenomena and Microfluidics session at the 82nd ACS Colloid and Surface Science Symposium in Raleigh, NC, in 2008, and which we hope will now be a regular feature at successive ACS Colloid and Surface Science meetings. This year at the combined 2009 13th International Conference on Surface and Colloid Science (ICSCS) and the 83rd ACS Colloid and Surface Science Symposium in New York, the Electrokinetics and Microfluidics symposium proved to be extremely popular, with three keynote lectures presented by Professor Howard Stone, Professor Hsueh-Chia Chang, and Professor Thomas Healy, and 44 oral presentations. In both 2008 and 2009, Biomicrofluidics has organized a special issue to cover some of the contributions reported at these meetings.2The growing interest in using electric fields to manipulate biological entities such as cells, DNA, and even single molecules is reflected in this year’s collection of papers with dielectrophoretic (DEP) phenomena comprising the bulk of the contributions. In Ref. 3, a new theory to describe Stern layer conductance along the surface of nanocolloids is proposed, forming the basis for the derivation of a more accurate prediction of the DEP crossover frequency. This theory is then employed to determine the conformation and, hence, optimum coverage of oligonucleotides on the surface of nanocolloid functionalized molecular probes during DNA hybridization under the influence of DEP, which can be exploited for biomolecular sensing. Other fundamental DEP papers include the investigation of particle motion under DEP induced optically via a photoconductor, in which Zhu et al.4 characterized the frequency dependence of the motion through the synchronous velocity spectra of the particles, and a numerical study of particle trapping at the throat of converging-diverging microchannels under the influence of negative DEP using a transient arbitrary Lagrangian–Eulerian finite element method.5 A more practical implementation is, on the other hand, reported by Yang et al.6 in which the negative DEP is exploited to separate colorectal cancer cells from other cells in a microfluidic device as a demonstration of a portable cancer detection tool.Continuing along the separation theme, but with regard to DNA separation using pulsed-field gel electrophoresis aided by sparse but regularly ordered microfabricated arrays of nanoposts, is a Brownian dynamics simulation model reported by Ou et al.7 in which DNA channeling, which predicts that the motion of DNA is undisturbed by the presence of arrays for large spacing to DNA equilibrium size ratios and when the field lines are straight, is predicted, consistent with experimental observations. In another fundamental paper, a direct numerical simulation model is presented to predict the current-voltage relationship across conducting pores along cell membranes, which is of fundamental importance in the electroporation process.8We hope that you will enjoy reading the contributions in this special topic and that it encourages you to participate in future Electrokinetics and Microfluidics meetings at the ACS Colloid and Surface Science Symposia, which we definitely hope will continue on a regular basis.  相似文献   

20.
A prerequisite for single cell study is the capture and isolation of individual cells. In microfluidic devices, cell capture is often achieved by means of trapping. While many microfluidic trapping techniques exist, hydrodynamic methods are particularly attractive due to their simplicity and scalability. However, current design guidelines for single cell hydrodynamic traps predominantly rely on flow resistance manipulation or qualitative streamline analysis without considering the target particle size. This lack of quantitative design criteria from first principles often leads to non-optimal probabilistic trapping. In this work, we describe an analytical design guideline for deterministic single cell hydrodynamic trapping through the optimization of streamline distributions under laminar flow with cell size as a key parameter. Using this guideline, we demonstrate an example design which can achieve 100% capture efficiency for a given particle size. Finite element modelling was used to determine the design parameters necessary for optimal trapping. The simulation results were subsequently confirmed with on-chip microbead and white blood cell trapping experiments.  相似文献   

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