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1.
Alternating-current (AC) electrokinetics involve the movement and behaviors of particles or cells. Many applications, including dielectrophoretic manipulations, are dependent upon charge interactions between the cell or particle and the surrounding medium. Medium concentrations are traditionally treated as spatially uniform in both theoretical models and experiments. Human red blood cells (RBCs) are observed to crenate, or shrink due to changing osmotic pressure, over 10 min experiments in non-uniform AC electric fields. Cell crenation magnitude is examined as functions of frequency from 250 kHz to 1 MHz and potential from 10 Vpp to 17.5 Vpp over a 100 μm perpendicular electrode gap. Experimental results show higher peak to peak potential and lower frequency lead to greater cell volume crenation up to a maximum volume loss of 20%. A series of experiments are conducted to elucidate the physical mechanisms behind the red blood cell crenation. Non-uniform and uniform electrode systems as well as high and low ion concentration experiments are compared and illustrate that AC electroporation, system temperature, rapid temperature changes, medium pH, electrode reactions, and convection do not account for the crenation behaviors observed. AC electroosmotic was found to be negligible at these conditions and AC electrothermal fluid flows were found to reduce RBC crenation behaviors. These cell deformations were attributed to medium hypertonicity induced by ion concentration gradients in the spatially nonuniform AC electric fields.  相似文献   

2.
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   

3.
Isolated mitochondria display a wide range of sizes plausibly resulting from the coexistence of subpopulations, some of which may be associated with disease or aging. Strategies to separate subpopulations are needed to study the importance of these organelles in cellular functions. Here, insulator-based dielectrophoresis (iDEP) was exploited to provide a new dimension of organelle separation. The dielectrophoretic properties of isolated Fischer 344 (F344) rat semimembranosus muscle mitochondria and C57BL/6 mouse hepatic mitochondria in low conductivity buffer (0.025–0.030 S/m) at physiological pH (7.2–7.4) were studied using polydimethylsiloxane (PDMS) microfluidic devices. First, direct current (DC) and alternating current (AC) of 0–50 kHz with potentials of 0–3000 V applied over a channel length of 1 cm were separately employed to generate inhomogeneous electric fields and establish that mitochondria exhibit negative DEP (nDEP). DEP trapping potential thresholds at 0–50 kHz were also determined to be weakly dependent on applied frequency and were generally above 200 V. Second, we demonstrated a separation scheme using DC potentials <100 V to perform the first size-based iDEP sorting of mitochondria. Samples of isolated mitochondria with heterogeneous sizes (150 nm–2 μm diameters) were successfully separated into sub-micron fractions, indicating the ability to isolate mitochondria into populations based on their size.  相似文献   

4.
To develop a lab on a chip (LOC) integrated with both sensor and actuator functions, a novel two-in-one system based on optical-driven manipulation and sensing in a microfluidics setup based on a hydrogenated amorphous silicon (a-Si:H) layer on an indium tin oxide/glass is first realized. A high-intensity discharge xenon lamp functioned as the light source, a chopper functioned as the modulated illumination for a certain frequency, and a self-designed optical path projected on the digital micromirror device controlled by the digital light processing module was established as the illumination input signal with the ability of dynamic movement of projected patterns. For light-addressable potentiometric sensor (LAPS) operation, alternating current (AC)-modulated illumination with a frequency of 800 Hz can be generated by the rotation speed of the chopper for photocurrent vs bias voltage characterization. The pH sensitivity, drift coefficient, and hysteresis width of the Si3N4 LAPS are 52.8 mV/pH, −3.2 mV/h, and 10.5 mV, respectively, which are comparable to the results from the conventional setup. With an identical two-in-one system, direct current illumination without chopper rotation and an AC bias voltage can be provided to an a-Si:H chip with a manipulation speed of 20 μm/s for magnetic beads with a diameter of 1 μm. The collection of magnetic beads by this light-actuated AC electroosmosis (LACE) operation at a frequency of 10 kHz can be easily realized. A fully customized design of an illumination path with less decay can be suggested to obtain a high efficiency of manipulation and a high signal-to-noise ratio of sensing. With this proposed setup, a potential LOC system based on LACE and LAPS is verified with the integration of a sensor and an actuator in a microfluidics setup for future point-of-care testing applications.  相似文献   

5.
Electrorotation is widely used for characterization of biological cells and materials using a rotating electric field. Generally, multiphase AC electric fields and quadrupolar electrode configuration are needed to create a rotating electric field for electrorotation. In this study, we demonstrate a simple method to rotate dielectrophoretically trapped microparticles using a stationary AC electric field. Coplanar interdigitated electrodes are used to create a linearly polarized nonuniform AC electric field. This nonuniform electric field is employed for dielectrophoretic trapping of microparticles as well as for generating electroosmotic flow in the vicinity of the electrodes resulting in rotation of microparticles in a microfluidic device. The rotation of barium titanate microparticles is observed in 2-propanol and methanol solvent at a frequency below 1 kHz. A particle rotation rate as high as 240 revolutions per minute is observed. It is demonstrated that precise manipulation (both rotation rate and equilibrium position) of the particles is possible by controlling the frequency of the applied electric field. At low frequency range, the equilibrium positions of the microparticles are observed between the electrode edge and electrode center. This method of particle manipulation is different from electrorotation as it uses induced AC electroosmosis instead of electric torque as in the case of electrorotation. Moreover, it has been shown that a microparticle can be rotated along its own axis without any translational motion.  相似文献   

6.
A biochip system imitates the oviduct of mammals with a microfluidic channel to achieve fertilization in vitro of imprinting-control-region (ICR) mice. We apply a method to manipulate and to position the oocyte and the sperm of ICR mice at the same time in our microfluidic channel with a positive dielectrophoretic (DEP) force. The positive dielectrophoretic response of the oocyte and sperm was exhibited under applied bias conditions AC 10 Vpp waveform, 1 MHz, 10 min. With this method, the concentration of sperm in the vicinity of the oocyte was increased and enhanced the probability of natural fertilization. We used commercial numerical software (CFDRC-ACE+) to simulate the square of the electric field and analyzed the location at which the oocyte and sperm are trapped. The microfluidic devices were designed and fabricated with poly(dimethylsiloxane). The results of our experiments indicate that a positive DEP served to drive the position of the oocyte and the sperm to natural fertilization (average rate of fertilization 51.58%) in our microchannel structures at insemination concentration 1.5 × 106 sperm ml−1. Embryos were cultured to two cells after 24 h and four cells after 48 h.  相似文献   

7.
This paper investigates the phenomenon of Faradaic charging in ac electrokinetics. Faradaic reactions were suggested as a key effect responsible for the reversal of pumping direction in ac micropumps. However, this hypothesis has yet to be proven convincingly and directly. Here we present an ion detection strategy to determine the production of ions through Faradaic hydrolytic reactions originating from direct application of voltage to electrolytic solutions during ac electrokinetics. Experiments were performed with symmetrical planar electrodes aligned along a microfluidic channel. Fluorescein, a pH-dependent dye, was employed as the pH indicator for the detection of ion production. Images were captured for analysis at various voltage levels. From analyzing the fluorescence intensity and its distribution, it can be concluded that the production of ions from hydrolytic reactions takes place and increases with the ac voltage. The coefficient of deviation indicates a significant enhancement at ac voltage above 11 Vpp. Lastly, we demonstrate a strategy using dc-biased ac electrokinetics to achieve controllability in direction and magnitude of the net fluid flow in pumping application.  相似文献   

8.
Spatially varied surface treatment of a fluorescently labeled Bovine Serum Albumin (BSA) protein, on the walls of a closed (sealed) microchannel is achieved via a well-defined gradient in plasma intensity. The microchips comprised a microchannel positioned in-between two microelectrodes (embedded in the chip) with a variable electrode separation along the length of the channel. The channel and electrodes were 50 μm and 100 μm wide, respectively, 50 μm deep, and adjacent to the channel for a length of 18 mm. The electrode separation distance was varied linearly from 50 μm at one end of the channel to a maximum distance of 150, 300, 500, or 1000 μm to generate a gradient in helium plasma intensity. Plasma ignition was achieved at a helium flow rate of 2.5 ml/min, 8.5 kVpk-pk, and 10 kHz. It is shown that the plasma intensity decreases with increasing electrode separation and is directly related to the residual amount of BSA left after the treatment. The plasma intensity and surface protein gradient, for the different electrode gradients studied, collapse onto master curves when plotted against electrode separation. This precise spatial control is expected to enable the surface protein gradient to be tuned for a range of applications, including high-throughput screening and cell-biomolecule-biomaterial interactions.  相似文献   

9.
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   

10.
Assessment of the dielectrophoresis (DEP) cross-over frequency (fxo), cell diameter, and derivative membrane capacitance (Cm) values for a group of undifferentiated human embryonic stem cell (hESC) lines (H1, H9, RCM1, RH1), and for a transgenic subclone of H1 (T8) revealed that hESC lines could not be discriminated on their mean fxo and Cm values, the latter of which ranged from 14 to 20 mF/m2. Differentiation of H1 and H9 to a mesenchymal stem cell-like phenotype resulted in similar significant increases in mean Cm values to 41–49 mF/m2 in both lines (p < 0.0001). BMP4-induced differentiation of RCM1 to a trophoblast cell-like phenotype also resulted in a distinct and significant increase in mean Cm value to 28 mF/m2 (p < 0.0001). The progressive transition to a higher membrane capacitance was also evident after each passage of cell culture as H9 cells transitioned to a mesenchymal stem cell-like state induced by growth on a substrate of hyaluronan. These findings confirm the existence of distinctive parameters between undifferentiated and differentiating cells on which future application of dielectrophoresis in the context of hESC manufacturing can be based.  相似文献   

11.
The capability of the AC dielectrophoresis (DEP) for on-chip capture and chaining of microalgae suspended in freshwaters was evaluated. The effects of freshwater composition as well as the electric field voltage, frequency, and duration, on the dielectrophoretic response of microalga Chlamydomonas reinhardtii were characterized systematically. Highest efficiency of cell alignment in one-dimensional arrays, determined by the percentage of cells in chain and the chain length, was obtained at AC-field of 20 V mm−1 and 1 kHz applied for 600 s. The DEP response and cell alignment of C. reinhardtii in water sampled from lake, pond, and river, as well as model media were affected by the chemical composition of the media. In the model media, the efficiency of DEP chaining was negatively correlated to the conductivity of the cell suspensions, being higher in suspensions with low conductivity. The cells suspended in freshwaters, however, showed anomalously high chaining at long exposure times. High concentrations of nitrate and dissolved organic matter decrease cell chaining efficiency, while phosphate and citrate concentrations increase it and favor formation of longer chains. Importantly, the application of AC-field had no effect on algal autofluorescence, cell membrane damage, or oxidative stress damages in C. reinhardtii.  相似文献   

12.
In this paper, the translational motion and self-rotational behaviors of the Raji cells, a type of B-cell lymphoma cell, in an optically induced, non-rotational, electric field have been characterized by utilizing a digitally programmable and optically activated microfluidics chip with the assistance of an externally applied AC bias potential. The crossover frequency spectrum of the Raji cells was studied by observing the different linear translation responses of these cells to the positive and negative optically induced dielectrophoresis force generated by a projected light pattern. This digitally projected spot served as the virtual electrode to generate an axisymmetric and non-uniform electric field. Then, the membrane capacitance of the Raji cells could be directly measured. Furthermore, Raji cells under this condition also exhibited a self-rotation behavior. The repeatable and controlled self-rotation speeds of the Raji cells to the externally applied frequency and voltage were systematically investigated and characterized via computer-vision algorithms. The self-rotational speed of the Raji cells reached a maximum value at 60 kHz and demonstrated a quadratic relationship with respect to the applied voltage. Furthermore, optically projected patterns of four orthogonal electrodes were also employed as the virtual electrodes to manipulate the Raji cells. These results demonstrated that Raji cells located at the center of the four electrode pattern could not be self-rotated. Instead any Raji cells that deviated from this center area would also self-rotate. Most importantly, the Raji cells did not exhibit the self-rotational behavior after translating and rotating with respect to the center of any two adjacent electrodes. The spatial distributions of the electric field generated by the optically projected spot and the pattern of four electrodes were also modeled using a finite element numerical simulation. These simulations validated that the electric field distributions were non-uniform and non-rotational. Hence, the non-uniform electric field must play a key role in the self-rotation of the Raji cells. As a whole, this study elucidates an optoelectric-coupled microfluidics-based mechanism for cellular translation and self-rotation that can be used to extract the dielectric properties of the cells without using conventional metal-based microelectrodes. This technique may provide a simpler method for label-free identification of cancerous cells with many associated clinical applications.  相似文献   

13.
Human mesenchymal stem cells (hMSCs) have three key properties that make them desirable for stem cell therapeutics: differentiation capacity, trophic activity, and ability to self-renew. However, current separation techniques are inefficient, time consuming, expensive, and, in some cases, alter hMSCs cellular function and viability. Dielectrophoresis (DEP) is a technique that uses alternating current electric fields to spatially separate biological cells based on the dielectric properties of their membrane and cytoplasm. This work implements the first steps toward the development of a continuous cell sorting microfluidic device by characterizing native hMSCs dielectric signatures and comparing them to hMSCs morphologically standardized with a polymer. A quadrapole Ti-Au electrode microdevice was used to observe hMSC DEP behaviors, and quantify frequency spectra and cross-over frequency of hMSCs from 0.010–35 MHz in dextrose buffer solutions (0.030 S/m and 0.10 S/m). This combined approach included a systematic parametric study to fit a core-shell model to the DEP spectra over the entire tested frequency range, adding robustness to the analysis technique. The membrane capacitance and permittivity were found to be 2.2 pF and 2.0 in 0.030 S/m and 4.5 pF and 4.1 in 0.10 S/m, respectively. Elastin-like polypeptide (ELP-) polyethyleneimine (PEI) copolymer was used to control hMSCs morphology to spheroidal cells and aggregates. Results demonstrated that ELP-PEI treatment controlled hMSCs morphology, increased experiment reproducibility, and concurrently increased hMSCs membrane permittivity to shift the cross-over frequency above 35 MHz. Therefore, ELP-PEI treatment may serve as a tool for the eventual determination of biosurface marker-dependent DEP signatures and hMSCs purification.  相似文献   

14.
A variety of methods have been used to introduce chemicals into a stream or to mix two or more streams of different compositions using microfluidic devices. In the following paper, the introduction of cryoprotective agents (CPAs) used during cryopreservation of cells in order to protect them from freezing injuries and increase viability post thaw is described. Dimethylsulphoxide (DMSO) is the most commonly used CPA. We aim to optimize the operating conditions of a two-stream microfluidic device to introduce a 10% vol/vol solution of DMSO into a cell suspension. Transport behavior of DMSO between two streams in the device has been experimentally characterized for a spectrum of flow conditions (0.7 < Re < 10), varying initial donor stream concentrations, (1% vol/vol < Co < 15% vol/vol) and different flow rate fractions (0.23 < fq < 0.77). The outlet cell stream concentration is analyzed for two different flow configurations: one with the cell stream flowing on top of the DMSO-rich donor stream, and the other with the cell stream flowing beneath the heavy DMSO-laden stream. We establish a transition from a diffusive mode of mass transfer to gravity-influenced convective currents for Atwood numbers (At) in the range of (1.7 × 10−3 < At < 3.1 × 10−3) for the latter configuration. Flow visualization with cells further our understanding of the effect of At on the nature of mass transport. Cell motion studies performed with Jurkat cells confirm a high cell recovery from the device while underscoring the need to collect both the streams at the outlet of the device and suggesting flow conditions that will help us achieve the target DMSO outlet concentration for clinical scale flow rates of the cell suspension.  相似文献   

15.
The time-dependent nucleation phase is critical to amyloid fibrillation and related to many pathologies, in which the conversion from natively folded amyloidogenic proteins to oligomers via nucleation is often hypothesized as a possible underlying mechanism. In this work, non-uniform AC-electric fields across two asymmetric electrodes were explored to control and examine the aggregation of insulin, a model amyloid protein, in aqueous buffer solution at constant temperature (20 °C) by fluorescence correlation spectroscopy and fluorescence microscopy. Insulin was rapidly concentrated in a strong AC-field by imposed AC-electroosmosis flow over an optimal frequency range of 0.5–2 kHz. In the presence of an AC-field, direct fibrillation from insulin monomers without the formation of oligomer precursors was observed. Once the insulin concentration had nearly doubled its initial concentration, insulin aggregates were observed in solution. The measured lag time for the onset of insulin aggregation, determined from the abrupt reduction in insulin concentration in solution, was significantly shortened from months or years in the absence of AC-fields to 1 min–3 h under AC-fields. The ability of external fields to alter amyloid nucleation kinetics provides insights into the onset of amyloid fibrillation.  相似文献   

16.
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17.
Genetic variation in the angiotensin II type 1 receptor (AT1R) has an important effect on the outcome of acute coronary syndrome (ACS) initiated treatment with captopril. This study aims to investigate the impact of genetic polymorphism of AT1R (rs5186 and rs275651) on the ACS outcome in Iraqi patients treated with captopril. A total of 250 Iraqi individuals with ACS were included in this case—control study and they were divided into two study groups; Study group 1 included 125 participants who were prescribed captopril, 25 mg twice daily and study group 2 included 125 participants who received no captopril as part of their ACS treatment (control study). The AT1R gene (rs5186) CC genotype was found to be associated with ST-elevation myocardial infarction (STEMI) (Odd’s ratio (O.R) = 1.2, P = 0.7), while AC was associated with Non-ST-elevation myocardial infarction (NSTEMI) and unstable angina (UA) (O.R = 1.2, P = 0.8). AC genotype is more prone to have Percutaneous coronary intervention (PCI) after ACS attack (O.R = 1.2, P = 0.6). CC genotype had a risk to get less improvement (O.R = 1.6, P = 0.5), so might require higher doses of captopril during acute coronary insult. The AT1R gene (rs275651) AA genotype was associated with UA (O.R = 1.3, P = 0.9). AA and AT genotypes were more prone to have PCI after ACS attack (O.R = 3.9 P = 0.2, O.R = 3.5, P = 0.3 respectively) and thus requiring higher doses of captopril. We conclude that the AT1R rs5186, rs275651 genetic polymorphisms might partially affect the clinical outcome of ACS patients treated with captopril and might have captopril resistance which requires higher doses.  相似文献   

18.
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   

19.
The authors exposed a non-equilibrium dynamic counterion and coion analyte concentration to an AC electric field to selectively concentrate peptides at the poles of a cation-selective granule. The counterion polarization results from the focusing of the electric field show a discontinuous drop in the intra-granule counterion electromigration flux at the pole. The coion concentration polarization is due to the combined external convective and electromigration fluxes toward the pole that neutralize the accumulating counterions. Because the electromigration mobility of the peptide anion analyte depends on the pH, the authors determined a 20 000-fold high concentration factor for a near-neutral pH of 6.0 to 7.7. Because the peptide is protonated at the acidic pole and its absolute charge ranges from −0.3 to −1.9, the concentration factor scales exponentially with the absolute charge, thus allowing extremely selective concentrations of various peptides, which is demonstrated by fluorescein isothiocyanate tagged angiotensin I (pI ∼ 5.8) and Texas red tagged avidin (pI ∼ 10.5). This dynamic concentration effect can substantially enhance the sensitivity of bio-assays.  相似文献   

20.
The selective cell separation is a critical step in fundamental life sciences, translational medicine, biotechnology, and energy harvesting. Conventional cell separation methods are fluorescent activated cell sorting and magnetic-activated cell sorting based on fluorescent probes and magnetic particles on cell surfaces. Label-free cell separation methods such as Raman-activated cell sorting, electro-physiologically activated cell sorting, dielectric-activated cell sorting, or inertial microfluidic cell sorting are, however, limited when separating cells of the same kind or cells with similar sizes and dielectric properties, as well as similar electrophysiological phenotypes. Here we report a label-free density difference amplification-based cell sorting (dDACS) without using any external optical, magnetic, electrical forces, or fluidic activations. The conceptual microfluidic design consists of an inlet, hydraulic jump cavity, and multiple outlets. Incoming particles experience gravity, buoyancy, and drag forces in the separation chamber. The height and distance that each particle can reach in the chamber are different and depend on its density, thus allowing for the separation of particles into multiple outlets. The separation behavior of the particles, based on the ratio of the channel heights of the inlet and chamber and Reynolds number has been systematically studied. Numerical simulation reveals that the difference between the heights of only lighter particles with densities close to that of water increases with increasing the ratio of the channel heights, while decreasing Reynolds number can amplify the difference in the heights between the particles considered irrespective of their densities.Separating specific cells from heterogeneous or homogeneous mixtures has been considered as a key step in a wide variety of applications ranging from biomedicine to energy harvesting. For example, the separation and sorting of rare circulating tumor cells (CTCs) from whole blood has gained significant importance in the potential diagnosis and treatment of metastatic cancers.1,2 Similarly, malaria detection relies on the collection of infected red blood cells (RBCs) from whole blood.3,4 In addition, the selective separation of lipid-rich microalgae from homogeneous mixtures of microalgae is a promising technique in biomass conversion.5To date, conventional cell separation can be done by labelling cells with biomolecules to induce differences in physical properties. For instance, in a fluorescence-activated cell sorter (FACS), cells to be separated are labelled with antibodies or aptamers with fluorescent molecules, and then sorted by applying an electrical potential.6,7 Similarly, magnetic-activated cell sorter (MACS) uses magnetic.8,9 Alternatively, label-free cell separation methods have exploited inherent differences in the physical properties (e.g., size and dielectric properties) of different kinds of cells. For example, acoustophoresis forces particles larger than a desired size to move into the center of a fluidic channel by using ultrasonic standing waves.10–12 Inertial microfluidics takes advantage of curved fluidic channels in order to amplify the size differences between particles.13,14 Mass-dependent separation of particles based on gravity and hydrodynamic flow was also reported.15 Particles with different dielectric properties can also be sorted by dielectrophoresis which induces the movement of polarizable particles.16–18The disadvantage of these methods, however, is that they require external forces and labels that may cause unexpected damage to biological cells.19–21 More importantly, most methods are limited in separating cells of the same kind or cells with similar sizes and dielectric properties.Here we designed a novel, label-free density difference amplification-based cell sorting (dDACS) that allows the separation of particles with the same size and charge by exploiting subtle differences in density without the use of external forces. Figure 1(a) illustrates the proposed microfluidic model and its underlying mechanism. The conceptual microfluidic system consists of an inlet, a separation chamber (hydraulic jump cavity), and multiple outlets. Particles entering through the inlet experience gravity (FG), buoyancy (FB), and drag (FD) forces in the separation chamber. The net force acting on the particles can be described as F = FG + FB + FD.(1)As particles enter the separation chamber (i.e., hydraulic jump cavity), FD acting on the particles changes its direction along the streamline. The particles experience additional forces in the y direction due to large tangential angle (Fig. 1(b)). For lighter particles, whose densities are close to that of the surrounding water, FD becomes comparable to FG (i.e., in the y direction), while the net force for heavier particles is less affected by this additional contribution of FD due to a large FG. As a result, the height (H) and distance (D) that each particle can travel are different depending on its density. The difference in the maximum height (ΔHmax) between two particles with different density (ρp1 and ρp2) can be further approximated as ΔHmax∝(vyp0)2(vyf−vyp0), (ρp1−ρp2) ,(2)where vyp0 and vyf represent the velocity of particle and fluid along the y direction at the entrance of hydraulic jump cavity, respectively.Open in a separate windowFIG. 1.Schematic illustration of label-free density difference amplification-based cell sorting (dDACS), which exploits differences in the densities (ρ1 > ρ2) of particles with similar diameters (d) and charge. (a) The conceptual microfluidic design consists of an inlet, a separation chamber (hydraulic jump cavity), and multiple outlets. Incoming particles experience gravity (FG), buoyancy (FB), and drag (FD) forces in the separation chamber, and depending on their densities, the height (H) and distance (D) that each particle is able to reach will be different, allowing the particles to be separated into multiple outlets. (b) Possible microfluidic channel configurations for density-based separation: Uniform channel height (left), gradual channel expansion (middle), and hydraulic jump cavity with sudden channel expansion (right). The height difference between particles with different densities can be amplified by the sudden channel expansion compared to the other two cases due to the relatively large tangential angle, θ of FD. (|θ1|≪ |θ2|) (see Fig. S1 in the supplementary material22).In comparison with the other two cases (Fig. 1(b) uniform channel height and gradual channel expansion), the height difference between the particles with different densities can be amplified by the sudden channel expansion in the hydraulic jump cavity due to relatively large tangential angle (see supplementary material22). Therefore, the particles can be separated through the multiple outlets, depending on their height and distance.In order to analyze the separation behavior of particles in the chamber according to differences in their densities, H and D are systematically investigated. The numerical simulations are performed using a commercial CFD software (CFX 14.0; ANSYS 14.0; ANSYS, Inc.). Particles with the same density may have different trajectories in the separation chamber depending on their inlet positions (Fig. 2(a)). Prior to this investigation, the maximum height (Hmax) and distance (Dmax) for each particle are compared by examining H and D of 100 identical particles at different inlet positions since the inlet position of particles could be controlled.20 Fig. 2(b) shows Hmax and Dmax of particles with respect to density at a fixed Reynolds number (Re = 0.1). Note that Reynolds number is defined as Re = ρfvfDh/μ, where ρf, vf, Dh, μ are density of fluid, velocity of the fluid, hydraulic diameter of a channel, and dynamic viscosity of the fluid, respectively. The hydraulic diameter in the Reynolds number is determined with the inlet channel. Particle densities in the range of 1.1 to 2.0 g/cm3 are chosen with the increase of 0.1 g/cm3. These values are quite reasonable in that the densities of many microorganisms such as microalgae are typically within this range and their densities can be varied by 0.2 g/m3 depending on their cellular context.23 The lighter particles travel with a higher Hmax, and longer Dmax. With the separation chamber, the height difference between particles with densities of 1.1 and 1.2 g/cm3 can be amplified by about 10 times as compared to that in a channel without the chamber, judging from the position where the 1.1 g/cm3 particle reaches its Hmax.Open in a separate windowFIG. 2.Microfluidic particle separation with respect to Reynolds number (Re). (a) Trajectories in the separation chamber of a hundred particles with the same density starting from inlet positions chosen arbitrarily in order to investigate the effect of the inlet positions on the maxima of the height (Hmax) and distance (Dmax) prior to further simulation. (b) Representative trajectories of particles having different densities from 1.1 to 2.0 g/cm3. (c) The maximum height (Hmax) of each particle with respect to Re. (d) Representative maximum distance (Dmax) of each particle at Re = 0.1. (Left) Streamline of fluid and representative trajectories of particles with densities of 1.1 and 2.0 g/cm3 in the separation chamber at Re = 0.1 (right).In Fig. 2(c), the values for Hmax of particles with respect to Reynolds number (Re) are presented. Since in our study, the maximum height (Hmax) and distance (Dmax) for each particle were compared by examining H and D of 100 identical particles that are randomly distributed in the channel (throughout all figures), there is little variation in Hmax and Dmax between each simulation. However, the standard deviation between each simulation is quite small and can be negligible. The Hmax values particles at Re = 0.5 with densities of 1.1 g/cm3 and 1.2 g/cm3 are 2.21 × 103 μm and 2.17 × 103 μm, respectively. The difference between Hmax of different particles, ΔHmax, increases with decreasing Re. For example, ΔHmax between particles with densities of 1.1 and 2.0 g/cm3 becomes 0.26 × 103 μm at Re = 1.0, but increases to 1.38 × 103 μm as Re decreases to 0.1. As Re increases (velocity of fluid increases), the relative velocity in the y direction between the fluid and the particle increases resulting in increasing of FD in the y direction since the velocity of particle in the y direction is very small at the entrance of the separation chamber. Thus, contribution of FD becomes comparable to the net force in the y direction. As a result, most of the particles even in the case of heavier ones travel quite similarly with the streamline, and ΔHmax subsequently decreases. On the other hand, as Re decreases, the contribution of FG becomes dominant due to the decrease of FD in the y direction. Consequently, the particles start to cross downwards streamlines as the density of the particles increases and Hmax gradually decreases. In addition, irrespective of their densities, ΔHmax of the particles increases with decreasing Re.Fig. 2(d) shows Dmax with respect to the density of the particles (left). Different densities of particles show different trajectories due to the relative contribution of FD to the net force in the y direction depending on the particle density (right). At Re = 0.1, Dmax of particles with densities of 1.1 cm3 and 1.2 g/cm3 are 2.91 × 104 μm and 1.43 × 104 μm, respectively. As the density of a particle increases, its Dmax dramatically decreases. The difference in Dmax between particles with densities of 1.1 and 1.2 g/cm3 is 1.48 × 104 μm, and 0.0037 × 104 μm for particles with densities of 1.9 and 2.0 g/cm3. The effect of FD is stronger compared to that of FG on lighter particles. Thus, lighter particles travel quite similarly with the streamline and finally have a large Dmax. On the other hand, heavier particles where effect of FG is stronger compared to that of FD cross downwards streamlines and finally have a small Dmax.Next, in order to investigate the separation behavior of particles with respect to the geometry of the microfluidic device, the effect of the ratio of the height of the separation chamber (hc) to the inlet (hi) on Hmax is investigated as shown in Fig. ​Fig.3.3. Interestingly, Hmax of particles with density of 1.1 g/cm3 increases from 1.93 × 103 μm to 6.48 × 103 μm while that of particles with density of 1.9 g/cm3 slightly changes from 0.70 × 103 μm to 0.73 × 103 μm as hc/hi increases from 5 to 20.Open in a separate windowFIG. 3.Microfluidic particle separation with respect to the ratio of the height of the inlet (hi) to the separation chamber (hc).This result can be attributed to two effects: (1) the change in the streamline and (2) the relative contribution of drag force to the net force depending on the density. With increasing hc/hi, dramatic increase in Hmax for lighter particles is because the streamline for the lighter ones experiences more vertical displacement in the separation chamber and the contribution of FD to the net force acting on the lighter one is more significant (see Fig. S2 in the supplementary material22).Based on this approach, we propose a microfluidic device for the selective separation of the lightest particle. Fig. 4(a) shows one unit (with three outlets) of the proposed microfluidic device that can be connected in series. The ratio of channel heights (hc/hi) is set to 20, and the particle densities are in the range of 1.1 ∼ 1.5 g/m3. Fig. 4(b) shows the representative separation behavior of the particles. A portion of the lightest particles (1.1 g/cm3) is selectively separated into the upper and middle outlets, while remaining light particles together with four other heavier particles with densities in the range of 1.2 to 1.5 g/cm3 leave through the lowest outlet. With a single operation of this unit, 40% of the lightest particles are recovered. In addition, the yield increases with increasing number of cycles (Fig. 4(c)).Open in a separate windowFIG. 4.(a) One unit of the proposed microfluidic device for the selective separation of the lightest particle based on the simulation results. Particles are separated into two outlets based on differences in both the height and distance travelled stemming from differences in density. (b) Representative separation behavior of particles observed in the device. (c) The yield of the lightest particle (1.1 g/cm3) with the proposed microfluidic device according to the number of cycles (i.e., this unit is assumed to be connected in series).In summary, we have demonstrated a label-free microfluidic system for the separation of particles according to subtle differences in their densities without external forces. Our microfluidic design consists simply of an inlet, a separation chamber, and multiple outlets. When entering the separation chamber, the particles experience an additional drag force in the y direction, amplifying the difference in both the height and the distance that the particles with different densities can travel within the chamber. At a fixed Reynolds number, with increasing particle density, Hmax decreases monotonously, and Dmax decreases dramatically. On the other hand, as Reynolds number increases, the difference between the heights of particles with different densities is attenuated. In addition, the simulation reveals that increasing the ratio of the channel heights increases the difference between the heights of particles only when their densities are close to that of the surrounding water. Based on this approach, a microfluidic device for the separation of the lightest particles has been proposed. We expect that our density-based separation design can be beneficial to the selective separation of specific microorganisms such as lipid-rich microalgae for energy harvesting application.  相似文献   

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