RESUMEN
Protein sequencing, as well as protein fingerprinting, has gained tremendous attention in the electrical sensing realm of solid-state nanopores and is challenging due to fast translocations and the use of high molar electrolytes. Despite providing an appreciable signal-to-noise ratio, high electrolyte concentrations can have adverse effects on the native protein structure. Herein, we present a thorough investigation of low electrolyte sensing conditions across a broad pH and voltage range generating conductive pulses (CPs) irrespective of protein net charge. We used Cas9 as the model protein and demonstrated that unfolding is noncooperative, represented by the gradual elongation or stretching of the protein, and sensitive to both the applied voltage and pH (i.e., charge state). The magnitude of unfolding and the isoelectric point (pI) of Cas9 was found to be correlated and a critical factor in our experiments. Electroosmotic flow (EOF) was always aligned with the transit direction, whereas electrophoretic force (EPF) was either reinforcing (pH < pI) or opposing (pH > pI) the protein's movement, which led to slower translocations at higher pH values. Further exploration of higher pH values led to slowing down of protein with > 30% of the population being slower than 0.5 ms. Our results would be critical for protein sensing at very low electrolytes and to retard their translocation speed without resorting to high-bandwidth equipment.
Asunto(s)
Proteína 9 Asociada a CRISPR/metabolismo , Nanoporos , Electroósmosis/instrumentación , Electroósmosis/métodos , Concentración de Iones de Hidrógeno , Punto Isoeléctrico , Conformación Proteica , Desplegamiento ProteicoRESUMEN
In this paper, a micromixer of a new configuration is presented, consisting of a spherical chamber in the center of which an ion-selective microsphere is placed. Stratified liquid is introduced through the chamber via inlet and outlet holes under an external pressure gradient and an external electric field is directed in such a way that the resulting electroosmotic flow is directed against the pressure-driven flow, resulting in mixing. The investigation is carried out by direct numerical simulation on a super-computer. Optimal values of the applied electric field are determined to yield strong mixing. Above this optimal mixing regime, a number of instabilities and bifurcations are realized, which qualitatively coincide with those occurring during electrophoresis of an ion-selective microgranule. As shown by our calculation, these instabilities do not lead to an enhanced mixing. The resulting electroconvective vortices remain confined near the surface of the microgranule, and do not sufficiently perturb the stratified fluid flow further from the granule. On the other hand, another type of instability caused by the salt concentration gradient can generate sufficiently strong oscillations to enhance mixing. However, this only occurs when the external electric field is sufficiently high that the electroosmotic flow is comparable to the pressure-driven flow. This ultimately leads to creation of reverse flows of the liquid and cessation of the device operation. Thus, it was shown that the best mixing occurs in the absence of electrokinetic instability. Based on the data obtained, it is possible to select the necessary geometric characteristics of the micromixer to achieve the optimal mixing mode for a given set of liquids, which may be ten times more effective than passive mixers at the same flow rates. A comparison with the experimental data of the other authors confirms the effectiveness of this device and its other capabilities. Furthermore, the basic device design can be operated in other modes, for example, an electrohydrodynamic pump, a streaming current generator, or even a micro-reactor, depending on the system parameters and choice of an ion-selective granule.
Asunto(s)
Electroósmosis , Modelos Químicos , Simulación por Computador , Electroósmosis/instrumentación , Electroforesis/instrumentación , Diseño de Equipo , Dispositivos Laboratorio en un Chip , MicroesferasRESUMEN
Vortex formation near a two-part cylinder with zeta potentials of different values but the same sign under an external DC electric field is numerically investigated in this paper. The cylinder, inserted in a straight microchannel filled with an aqueous solution, is composed of an upstream part and a downstream part. When a DC electric field is applied in the channel, under certain conditions, the vortex will form near the cylinder due to the different velocities of electroosmotic flow generated on the cylinder surface. The numerical results reveal that the larger the velocity difference of electroosmotic flow generated on the two-part cylinder and the smaller the channel width, the more conducive to vortex formation in the channel. In addition, if the zeta potential ratios of cylinder downstream part to upstream part and channel wall to cylinder upstream part are unchanged, the DC electric field strength and the zeta potential value do not affect the pattern of vortices formed in the channel. This study provides a way for vortex formation in microchannels and has the potential application in microfluidic devices.
Asunto(s)
Electroósmosis/instrumentación , Microfluídica/instrumentación , Simulación por Computador , Diseño de Equipo , Modelos Químicos , Propiedades de SuperficieRESUMEN
The study of heat and electroosmotic characteristics in the flow of a third-order fluid regulated by peristaltic pumping is examined by using governing equations, i.e., the continuity equation, momentum equation, energy equation, and concentration equation. The wavelength is considered long compared to its height and a low Reynolds number is assumed. The velocity slip condition is employed. Analytical solutions are performed through the perturbation technique. The expressions for the dimensionless velocity components, temperature, concentration, and heat transfer rate are obtained. Pumping features were computed numerically for discussion of results. Trapping and heat transfer coefficient distributions were also studied graphically. The findings of the present study can be applied to design biomicrofluidic devices like tumor-on-a-chip and organ-on-a-chip.
Asunto(s)
Electroósmosis/instrumentación , Calor , Hidrodinámica , Modelos TeóricosRESUMEN
Electroosmotic flow (EOF) has been widely used to transport fluids and samples in micro- and nanofluidic channels for lab-on-a-chip applications. This essentially surface-driven plug-like flow is, however, sensitive to both the fluid and wall properties, of which any inhomogeneity may draw disturbances to the flow and even instabilities. Existing studies on EOF instabilities have been focused primarily upon Newtonian fluids though many of the chemical and biological solutions are actually non-Newtonian. We carry out a systematic experimental investigation of the fluid rheological effects on the elastic instability in the EOF of phosphate buffer-based polymer solutions through T-shaped microchannels. We find that electro-elastic instabilities can be induced in shear thinning polyacrylamide (PAA) and xanthan gum (XG) solutions if the applied direct current voltage is above a threshold value. However, no instabilities are observed in Newtonian or weakly shear thinning viscoelastic fluids including polyethylene oxide (PEO), polyvinylpyrrolidone (PVP), and hyaluronic acid (HA) solutions. We also perform a quantitative analysis of the wave parameters for the observed elasto-elastic instabilities.
Asunto(s)
Electroósmosis/instrumentación , Técnicas Analíticas Microfluídicas/instrumentación , Modelos Químicos , Resinas Acrílicas/química , Elasticidad , Reología , ViscosidadRESUMEN
Quantitative mass spectrometry analysis of metabolites at a single-cell level is critical to understanding the cell functionality and heterogeneity. To preserve cell viability after extraction, the extracted volume needs to be precisely controlled at a subpicoliter-to-picoliter level. Recently, we developed a volume-controlled, and highly sensitive approach for live cell analysis at a single-cell level by integrating electroosmotic extraction and nano-electrospray ionization mass spectrometry (nanoESI MS) analysis. Herein, we use outer epidermal cells of Allium cepa as a model system to present the details of our workflow, including detailed descriptions of the experimental setup for live cell analysis, preparation of the extraction nanopipette, establishment of calibration curves, and extraction and quantification of glucose in an individual onion cell. The capability of this procedure for quantitative live cell analysis has been demonstrated by accurate quantification of glucose in Allium cepa. In principle, our approach is applicable to identification and quantification of metabolites in live mammalian cells.
Asunto(s)
Electroósmosis/instrumentación , Cebollas/citología , Células Vegetales/metabolismo , Espectrometría de Masa por Ionización de Electrospray/instrumentación , Supervivencia Celular , Electroósmosis/métodos , Diseño de Equipo , Glucosa/análisis , Glucosa/metabolismo , Metabolómica/instrumentación , Metabolómica/métodos , Cebollas/química , Cebollas/metabolismo , Células Vegetales/química , Análisis de la Célula Individual/instrumentación , Análisis de la Célula Individual/métodos , Espectrometría de Masa por Ionización de Electrospray/métodosRESUMEN
Plant polyphenols can form functional coatings on various materials through self-polymerization. In this paper, a series of modified capillary columns, which possess diversity of charge characteristics for modulating electroosmotic flow (EOF), were prepared by one-step co-deposition of gallic acid (GA), a plant-derived polyphenol monomer, and branched polyethyleneimine (PEI). The physicochemical properties of the prepared columns were characterized by Fourier transform infrared spectroscopy (FT-IR), UV-Vis spectroscopy and scanning electron microscopy (SEM). The magnitude and direction of EOF of GA/PEI co-deposited columns were modulated by changing a series of coating parameters, such as post-incubation of FeCl3, co-deposition time, and deposited amounts of GA and PEI with different relative molecular mass (PEI-600, PEI-1800, PEI-10000, and PEI-70000). Furthermore, the separation efficiencies of the prepared GA/PEI co-deposited columns were evaluated by separations of small molecules, including organic acids, polar nucleotides, phenols, nucleic acid bases and nucleosides. Results indicated that modulating of EOF plays an important role in enhancing the separation performance and reversing the elution order of the analytes. Finally, the developed method was successfully applied to quantitative analysis of acidic compounds in four real samples. The recoveries were in the range of 73.5%-85.8% for citric acid, benzoic acid, sorbic acid, salicylic acid and ascorbic acid in beverage and fruit samples, 101.6%-104.9% for cinnamic acid, vanillic acid, and ferulic acid in Angelica sinensis sample, while 84.6%-97.8% for guanosine-5'-monophosphate, uridine-5'-monophosphate, cytosine-5'- monophosphate and adenosine-5'-monophosphate in Cordyceps samples. These results indicated that the co-deposition of plant polyphenol-inspired GA/PEI coatings can provide new opportunities for EOF modulation of capillary electrophoresis.
Asunto(s)
Electrocromatografía Capilar/métodos , Electroósmosis/métodos , Ácido Gálico/química , Polietileneimina/química , Electrocromatografía Capilar/instrumentación , Electroósmosis/instrumentación , Peso Molecular , Ácidos Nucleicos/aislamiento & purificación , Nucleósidos/aislamiento & purificación , Nucleótidos/aislamiento & purificación , Compuestos Orgánicos/aislamiento & purificación , PolimerizacionRESUMEN
In this work, an efficient electroosmotic pump (EOP) based on the ultrathin silica nanoporous membrane (u-SNM), which can drive the motion of fluid under the operating voltage as low as 0.2 V, has been fabricated. Thanks to the ultrathin thickness of u-SNM (â¼75 nm), the effective electric field strength across u-SNM could be as high as 8.27 × 105 V m-1 in 0.4 M KCl when 1.0 V of voltage was applied. The maximum normalized electroosmotic flow (EOF) rate was as high as 172.90 mL/min/cm2 /V, which was larger than most of other nanoporous membrane based EOPs. In addition to the ultrathin thickness, the high porosity of this membrane (with a pore density of 4 × 1012 cm-2 , corresponding to a porosity of 16.7%) also contribute to such a high EOF rate. Moreover, the EOF rate was found to be proportional to both the applied voltage and the electrolyte concentration. Because of small electrokinetic radius of u-SNM arising from its ultrasmall pore size (ca. 2.3 nm in diameter), the EOF rate increased with increasing the electrolyte concentration and reached the maximum at a concentration of 0.4 M. This dependence was rationalized by the variations of both zeta potential and electrokinetic radius with the electrolyte concentration.
Asunto(s)
Electroósmosis/instrumentación , Membranas Artificiales , Nanoporos , Dióxido de Silicio/química , Electroósmosis/métodos , Diseño de Equipo , Modelos Químicos , PorosidadRESUMEN
Electrophoretic on-line sample preconcentration techniques in microfluidic channels improve the sensitivity prior to the separation. Among various techniques, the most important field-amplified sample stacking and sweeping on cross-channel microchips are demonstrated. As a novel microfluidic preconcentration approach, a large-volume sample stacking with electroosmotic flow pump (LVSEP) on straight-channel chips is also presented, which can omit a complicated voltage program for sample injection processes. In this chapter, we describe how to prepare and how to run these on-line sample preconcentration methods in microchip electrophoresis.
Asunto(s)
Electroósmosis/instrumentación , Electroforesis por Microchip/métodos , Tampones (Química) , Calibración , Electroósmosis/métodos , Electroforesis por Microchip/instrumentación , Límite de Detección , Técnicas Analíticas Microfluídicas/instrumentación , Técnicas Analíticas Microfluídicas/métodosRESUMEN
Insulator-based dielectrophoresis has to date been almost entirely restricted to Newtonian fluids despite the fact that many of the chemical and biological fluids exhibit non-Newtonian characteristics. We present herein an experimental study of the fluid rheological effects on the electroosmotic flow of four types of polymer solutions, i.e., 2000 ppm xanthan gum (XG), 5% polyvinylpyrrolidone (PVP), 3000 ppm polyethylene oxide (PEO), and 200 ppm polyacrylamide (PAA) solutions, through a constriction microchannel under DC electric fields of up to 400 V/cm. We find using particle streakline imaging that the fluid elasticity does not change significantly the electroosmotic flow pattern of weakly shear-thinning PVP and PEO solutions from that of a Newtonian solution. In contrast, the fluid shear-thinning causes multiple pairs of flow circulations in the weakly elastic XG solution, leading to a central jet with a significantly enhanced speed from before to after the channel constriction. These flow vortices are, however, suppressed in the strongly viscoelastic and shear-thinning PAA solution.
Asunto(s)
Electroósmosis/métodos , Soluciones/química , Resinas Acrílicas/química , Elasticidad , Electroósmosis/instrumentación , Técnicas Analíticas Microfluídicas/instrumentación , Polietilenglicoles/química , Polisacáridos Bacterianos/química , Povidona/química , Sustancias Viscoelásticas/químicaRESUMEN
Induced-charge electroosmosis (ICEO) has attracted tremendous popularity for driving fluid motion from the microfluidic community since the last decade, while less attention has been paid to ICEO-based nanoparticle manipulation. We propose herein a unique concept of hybrid electroosmotic kinetics (HEK) in terms of bi-phase ICEO (BICEO) actuated in a four-terminal spiral electrode array, for effective electrokinetic enrichment of fluorescent polystyrene nanoparticles on ideally polarizable metal strips. First, by alternating the applied AC voltage waves between consecutive discrete terminals, the flow stagnation lines where the sample nanoparticles aggregate can be switched in time between two different distribution modes. Second, we innovatively introduce the idea of AC field-effect flow control on BICEO; by altering the combination of gating voltage sequence, not only the number of circulative particle trapping lines is doubled, but the collecting locations can be flexibly reconfigured as well. Third, hydrodynamic streaming of DC-biased BICEO is tested in our device design, wherein the global linear electroosmosis dominates BICEO contributed from both AC and DC components, resulting in a reduction of particle enrichment area, while with a sharp increase in sample transport speed inside the bulk phase. The flow field associated with HEK is predicted using a linear asymptotic analysis under Debye-Huckel limit, with the simulation results in qualitative agreement with in-lab observations of nanoparticle trapping by exploiting a series of improved ICEO techniques. This work provides an affordable and field-deployable platform for real-time nanoparticle trapping in the context of dilute electrolyte.
Asunto(s)
Electroósmosis/instrumentación , Técnicas Analíticas Microfluídicas/instrumentación , Nanopartículas/química , Electricidad , Electroósmosis/métodos , Diseño de Equipo , Cinética , MicroelectrodosRESUMEN
Dielectrophoresis (DEP) is usually effective close to the electrode surface. Several techniques have been developed to overcome its drawbacks and to enhance dielectrophoretic particle capture. Here we present a simple technique of superimposing alternating current DEP (high-frequency signals) and electroosmosis (EO; low-frequency signals) between two coplanar electrodes (gap: 25 µm) using a lab-made voltage adder for rapid and selective concentration of bacteria, viruses, and proteins, where we controlled the voltages and frequencies of DEP and EO separately. This signal superimposition technique enhanced bacterial capture (Escherichia coli K-12 against 1-µm-diameter polystyrene beads) more selectively (>99%) and rapidly (~30 s) at lower DEP (5 Vpp) and EO (1.2 Vpp) potentials than those used in the conventional DEP capture studies. Nanometer-sized MS2 viruses and troponin I antibody proteins were also concentrated using the superimposed signals, and significantly more MS2 and cTnI-Ab were captured using the superimposed signals than the DEP (10 Vpp) or EO (2 Vpp) signals alone (p < 0.035) between the two coplanar electrodes and at a short exposure time (1 min). This technique has several advantages, such as simplicity and low cost of electrode fabrication, rapid and large collection without electrolysis.
Asunto(s)
Electroósmosis/instrumentación , Escherichia coli K12/aislamiento & purificación , Levivirus/aislamiento & purificación , Poliestirenos/química , Proteínas/aislamiento & purificación , Electricidad , Electrodos , Diseño de EquipoRESUMEN
This study uses negative dielectrophoresis and AC electroosmosis as a driving mechanism and presents an electrically driven microconcentrator that concentrates the sample in the region exterior to the electrodes (termed as exterior-electrode electrically driven microconcentrator in this paper). The proposed microconcentrator uses a 3-D face-to-face electrode pair; the top electrode is a relatively large planar electrode, and the bottom electrode is formed with three to six long and thin electrodes connected into an open ring. The sample is brought to the vicinity of the open electrode at the bottom by electroosmotic flow; then, negative dielectrophoresis is used to push the sample away from the electrode and concentrate it in the region surrounded by the open ring electrode. Concentration using an exterior-electrode electrically driven microconcentrator offers promise for convenient use in conjunction with relevant detection systems. The results indicate that for the proposed exterior-electrode electrically driven microconcentrator, the optimal frequency is 100 kHz and the optimal voltage is 13 Vp-p . The corner concentration process at the corners of the bottom open electrodes enables the multi-corner electrodes to exhibit better concentration results than that exhibited by semicircular-shaped electrodes. The concentration performance is most favorable when the shape of the open electrode at the bottom is a five-vertex electrode, enabling a concentration enhancement factor of 55 times for a latex particle solution and 11 times for E. coli. The experimental results also demonstrate that the concentration phenomenon in this study is not induced by non-specific adsorption and can be repeated multiple times.
Asunto(s)
Electroósmosis/instrumentación , Electroforesis/instrumentación , Electrodos , Diseño de Equipo , Escherichia coli/aislamiento & purificación , Microesferas , Modelos QuímicosRESUMEN
A nanoporous poly-(styrene sulfonate) (poly-SS) membrane was developed for fast and selective ion transport in a microfluidic chip. The poly-SS membrane can be photopolymerized in-situ at arbitrary location of a microchannel, enabling integrated fluidics design in the microfluidic chip. The membrane is characterized by a low hydraulic resistance and a high surface charge to maximize the electroosmotic flow and charge selectivity. The membrane characteristics were investigated by charge-selective electropreconcentration method. Experimental results show membranes with various percentages of poly-SS are able to concentrate anions (fluorescein and TRITC-labeled BSA). The anion-selective electropreconcentration process is stable and 26-times faster than previously reported poly-AMPS (2-acrylamido-2-methyl-1-propanesulfonic acid) based system. The electropreconcentration was also demonstrated to depend on the sample valency and buffer concentration.
Asunto(s)
Electroósmosis/métodos , Membranas Artificiales , Técnicas Analíticas Microfluídicas/métodos , Nanoporos , Aniones/análisis , Aniones/química , Aniones/aislamiento & purificación , Electroósmosis/instrumentación , Diseño de Equipo , Técnicas Analíticas Microfluídicas/instrumentación , Poliestirenos/químicaRESUMEN
We present a novel microfluidic paper-based analytical device (µPAD) which utilizes the native high electroosmotic flow (EOF) in nitrocellulose to achieve stationary isotachophoresis (ITP) focusing. This approach decouples sample accumulation from the length of the channel, resulting in significant focusing over short channel lengths. We provide a brief theory for EOF-balanced ITP focusing under continuous injection from a depleting reservoir and present the design of a short (7 mm) paper-based microfluidic channel, which allows a 200 µL sample to be processed in approximately 6 min, resulting in a 20 000-fold increase in concentration - a full order of magnitude improvement compared to previous paper-based ITP devices. We show the stability of the assay over longer (40 min) durations of time, and using Morpholino probes, we present the applicability of the device for amplification-free detection of nucleic acids, with a limit-of-detection (LoD) of 5 pM in 10 min. Finally, we utilize the small footprint of the channel and show a multiplexed platform in which 12 assays operate in parallel in a 24-well plate format.
Asunto(s)
ADN/análisis , Electroósmosis , Isotacoforesis , Técnicas Analíticas Microfluídicas , Papel , Colodión/química , Electroósmosis/instrumentación , Isotacoforesis/instrumentación , Técnicas Analíticas Microfluídicas/instrumentaciónRESUMEN
Insulator-based dielectrophoresis (iDEP) exploits in-channel hurdles and posts etc. to create electric field gradients for various particle manipulations. However, the presence of such insulating structures also amplifies the Joule heating in the fluid around themselves, leading to both temperature gradients and electrothermal flow. These Joule heating effects have been previously demonstrated to weaken the dielectrophoretic focusing and trapping of microscale and nanoscale particles. We find that the electrothermal flow vortices are able to entrain submicron particles for a localized enrichment near the insulating tips of a ratchet microchannel. This increase in particle concentration is reasonably predicted by a full-scale numerical simulation of the mass transport along with the coupled charge, heat and fluid transport. Our model also predicts the electric current and flow pattern in the fluid with a good agreement with the experimental observations.
Asunto(s)
Electroósmosis/instrumentación , Electroforesis/instrumentación , Técnicas Analíticas Microfluídicas/métodos , Microesferas , Dimetilpolisiloxanos/química , Electricidad , Campos Electromagnéticos , Diseño de Equipo/instrumentación , Concentración de Iones de Hidrógeno , Modelos Teóricos , Propiedades de Superficie , Temperatura , TermodinámicaRESUMEN
We report herein a novel microfluidic particle concentrator that utilizes constriction microchannels to enhance the flow-focusing performance of induced-charge electroosmosis (ICEO), where viscous hemi-spherical oil droplets are embedded within the mainchannel to form deformable converging-diverging constriction structures. The constriction region between symmetric oil droplets partially coated on the electrode strips can improve the focusing performance by inducing a granular wake flow area at the diverging channel, which makes almost all of the scattered sample particles trapped within a narrow stream on the floating electrode. Another asymmetric droplet pair arranged near the outlets can further direct the trajectory of focused particle stream to one specified outlet port depending on the symmetry breaking in the shape of opposing phase interfaces. By fully exploiting rectification properties of induced-charge electrokinetic phenomena at immiscible water/oil interfaces of tunable geometry, the expected function of continuous and switchable flow-focusing is demonstrated by preconcentrating both inorganic silica particles and biological yeast cells. Physical mechanisms responsible for particle focusing and locus deflection in the droplet-assisted concentrentor are analyzed in detail, and simulation results are in good accordance with experimental observations. Our work provides new routes to construct flexible electrokinetic framework for preprocessing on-chip biological samples before performing subsequent analysis.
Asunto(s)
Electroósmosis/instrumentación , Técnicas Analíticas Microfluídicas/instrumentación , Diseño de Equipo , Dióxido de Silicio , Levaduras/citologíaRESUMEN
Employing electric phenomena for the spatial manipulation of bioparticles from whole cells down to dissolved molecules has become a useful tool in biotechnology and analytics. AC electrokinetic effects like dielectrophoresis and AC electroosmosis are increasingly used to concentrate, separate and immobilize DNA and proteins. With the advance of photolithographical micro- and nanofabrication methods, novel or improved bioanalytical applications benefit from concentrating analytes, signal enhancement and locally controlled immobilization by AC electrokinetic effects. In this review of AC electrokinetics of proteins, the respective studies are classified according to their different electrode geometries: individual electrode pairs, interdigitated electrodes, quadrupole electrodes, and 3D configurations of electrode arrays. Known advantages and disadvantages of each layout are discussed.
Asunto(s)
Electroósmosis/instrumentación , Electroforesis/instrumentación , Proteínas/análisis , Animales , Electricidad , Electrodos , Electroósmosis/métodos , Electroforesis/métodos , Diseño de Equipo , HumanosRESUMEN
This paper presents the use of DC-biased AC electric field for enhancing cell trapping throughput in an insulator-based dielectrophoretic (iDEP) fluidic device with densely packed silica beads. Cell suspension is carried through the iDEP device by a pressure-driven flow. Under an applied DC-biased AC electric field, DEP trapping force is produced as a result of non-uniform electric field induced by the gap of electrically insulating silica beads packed between two mesh electrodes that allow both fluid and cells to pass through. While the AC component is mainly to control the magnitude of DEP trapping force, the DC component generates local electroosmotic (EO) flow in the cavity between the beads and the EO flow can be set to move along or against the main pressure-driven flow. Our experimental and simulation results show that desirable trapping is achieved when the EO flow direction is along (not against) the main flow direction. Using our proposed DC-biased AC field, the device can enhance the trapping throughput (in terms of the flowrate of cell suspension) up to five times while yielding almost the same cell capture rates as compared to the pure AC field case. Additionally, the device was demonstrated to selectively trap dead yeast cells from a mixture of flowing live and dead yeast cells.
Asunto(s)
Electroósmosis/instrumentación , Electroforesis/instrumentación , Ensayos Analíticos de Alto Rendimiento/instrumentación , Saccharomyces cerevisiae/aislamiento & purificación , Dióxido de Silicio/química , Adhesión Celular , Electricidad , Electrodos , Campos Electromagnéticos , Diseño de Equipo/instrumentación , Modelos Teóricos , Presión , Propiedades de SuperficieRESUMEN
Single-cell analysis has attracted increasing attention because of cell heterogeneities. Various strategies have been developed for analyzing single cells, but most of these analytical processes kill the cells. Tools that can qualitatively and quantitatively measure the cellular contents without killing the cell are highly demanding because they enable us to conduct single-cell time-course studies (e.g., to examine how a cell responds to a therapy before, during, and after a treatment). Here we develop a femto-liter (fL) pipet to serve this purpose. To ensure that we can accurately and precisely pipet fL solutions, we fill all conduits with liquid and use an electroosmotic pump (EOP) as the driving force to facilitate withdrawal of cellular contents from single cells. We tentatively term this device an EOP-driven pipette or EDP. We characterize the EDP for accurately and precisely withdrawing solution from â¼250 fL to 80 nL; a volume range that covers the applications for most types of cells. To demonstrate the feasibility of utilizing the EDP for a single-cell time-course study, we utilize the EDP to take the cellular contents out at different times during the course of a zebrafish embryo development for cholesterol measurements. More than 50% of the embryos survive after each pipetting and analysis step, and this number will increase considerably as we improve our cell manipulation skills and reduce the pipet-tip diameter. We expect this EDP to become an effective tool for single-cell time-course studies.