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1.
Deterministic lateral displacement (DLD) is a microfluidic size-based particle separation or filter technology with applications in cell separation and enrichment. Currently, there are no cost-effective manufacturing methods for this promising microfluidic technology. In this fabrication paper, however, we develop a simple, yet robust protocol for thermoplastic DLD devices using regulatory-approved materials and biocompatible methods. The final standalone device allowed for volumetric flow rates of 660 μl min−1 while reducing the manufacturing time to <1 h. Optical profilometry and image analysis were employed to assess manufacturing accuracy and precision; the average replicated post height was 0.48% less than the average post height on the master mold and the average replicated array pitch was 1.1% less than the original design with replicated posts heights of 62.1 ± 5.1 μm (mean ± 6 standard deviations) and replicated array pitches of 35.6 ± 0.31 μm. 相似文献
2.
Separation by dielectrophoresis of dormant and nondormant bacterial cells of Mycobacterium smegmatis
The dielectrophoretic behavior of active, dead, and dormant Mycobacterium smegmatis bacterial cells was studied. It was found that the 72-h-old dormant cells had a much higher effective particle conductivity (812±10 μS cm−1), almost double that of active cells (560±20 μS cm−1), while that of dead (autoclaved) M. smegmatis cells was the highest (950±15 μS cm−1) overall. It was also found that at 80 kHz, 900 μS cm−1 dead cells were attracted at the edges of interdigitated castellated electrodes by positive dielectrophoresis, but dormant cells were not. Similarly, at 120 kHz, 2 μS cm−1 active cells were attracted and dormant cells were not. Using these findings a dielectrophoresis-based microfluidic separation system was developed in which dead and active cells were collected from a given cell suspension, while dormant cells were eluted. 相似文献
3.
Understanding the correlation between exposed surfaces and performances of controlled nanocatalysts can aid effective strategies to enhance electrocatalysis, but this is as yet unexplored for the nitrogen reduction reaction (NRR). Here, we first report controlled synthesis of well-defined Pt3Fe nanocrystals with tunable morphologies (nanocube, nanorod and nanowire) as ideal model electrocatalysts for investigating the NRR on different exposed facets. The detailed electrocatalytic studies reveal that the Pt3Fe nanocrystals exhibit shape-dependent NRR electrocatalysis. The optimized Pt3Fe nanowires bounded with high-index facets exhibit excellent selectivity (no N2H4 is detected), high activity with NH3 yield of 18.3 μg h−1 mg−1cat (0.52 μg h−1 cm−2ECSA; ECSA: electrochemical active surface area) and Faraday efficiency of 7.3% at −0.05 V versus reversible hydrogen electrode, outperforming the {200} facet-enclosed Pt3Fe nanocubes and {111} facet-enclosed Pt3Fe nanorods. They also show good stability with negligible activity change after five cycles. Density functional theory calculations reveal that, with high-indexed facet engineering, the Fe-3d band is an efficient d-d coupling correlation center for boosting the Pt 5d-electronic exchange and transfer activities towards the NRR. 相似文献
4.
Accurate measurement of blood viscoelasticity including viscosity and elasticity is essential in estimating blood flows in arteries, arterials, and capillaries and in investigating sub-lethal damage of RBCs. Furthermore, the blood viscoelasticity could be clinically used as key indices in monitoring patients with cardiovascular diseases. In this study, we propose a new method to simultaneously measure the viscosity and elasticity of blood by simply controlling the steady and transient blood flows in a microfluidic analogue of Wheastone-bridge channel, without fully integrated sensors and labelling operations. The microfluidic device is designed to have two inlets and outlets, two side channels, and one bridge channel connecting the two side channels. Blood and PBS solution are simultaneously delivered into the microfluidic device as test fluid and reference fluid, respectively. Using a fluidic-circuit model for the microfluidic device, the analytical formula is derived by applying the linear viscoelasticity model for rheological representation of blood. First, in the steady blood flow, the relationship between the viscosity of blood and that of PBS solution (μBlood/μPBS) is obtained by monitoring the reverse flows in the bridge channel at a specific flow-rate rate (QPBSSS/QBloodL). Next, in the transient blood flow, a sudden increase in the blood flow-rate induces the transient behaviors of the blood flow in the bridge channel. Here, the elasticity (or characteristic time) of blood can be quantitatively measured by analyzing the dynamic movement of blood in the bridge channel. The regression formula (ABlood (t) = Aα + Aβ exp [−(t − t0)/λBlood]) is selected based on the pressure difference (ΔP = PA − PB) at each junction (A, B) of both side channels. The characteristic time of blood (λBlood) is measured by analyzing the area (ABlood) filled with blood in the bridge channel by selecting an appropriate detection window in the microscopic images captured by a high-speed camera (frame rate = 200 Hz, total measurement time = 7 s). The elasticity of blood (GBlood) is identified using the relationship between the characteristic time and the viscosity of blood. For practical demonstrations, the proposed method is successfully applied to evaluate the variations in viscosity and elasticity of various blood samples: (a) various hematocrits form 20% to 50%, (b) thermal-induced treatment (50 °C for 30 min), (c) flow-induced shear stress (53 ± 0.5 mL/h for 120 min), and (d) normal rat versus spontaneously hypertensive rat. Based on these experimental demonstrations, the proposed method can be effectively used to monitor variations in viscosity and elasticity of bloods, even with the absence of fully integrated sensors, tedious labeling and calibrations. 相似文献
5.
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
(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
6.
Gergely Simon Caroline Busch Marco A. B. Andrade Julien Reboud Jonathan M. Cooper Marc P. Y. Desmulliez Mathis O. Riehle Anne L. Bernassau 《Biomicrofluidics》2021,15(1)
Separation and sorting of biological entities (viruses, bacteria, and cells) is a critical step in any microfluidic lab-on-a-chip device. Acoustofluidics platforms have demonstrated their ability to use physical characteristics of cells to perform label-free separation. Bandpass-type sorting methods of medium-sized entities from a mixture have been presented using acoustic techniques; however, they require multiple transducers, lack support for various target populations, can be sensitive to flow variations, or have not been verified for continuous flow sorting of biological cells. To our knowledge, this paper presents the first acoustic bandpass method that overcomes all these limitations and presents an inherently reconfigurable technique with a single transducer pair for stable continuous flow sorting of blood cells. The sorting method is first demonstrated for polystyrene particles of sizes 6, 10, and 14.5 μm in diameter with measured purity and efficiency coefficients above 75 ± 6% and 85 ± 9%, respectively. The sorting strategy was further validated in the separation of red blood cells from white blood cells and 1 μm polystyrene particles with 78 ± 8% efficiency and 74 ± 6% purity, respectively, at a flow rate of at least 1 μl/min, enabling to process finger prick blood samples within minutes. 相似文献
7.
Zhengwei Zhang Peng Chen Xiangdong Yang Yuan Liu Huifang Ma Jia Li Bei Zhao Jun Luo Xidong Duan Xiangfeng Duan 《国家科学评论(英文版)》2020,7(4):737
Monolayer transition metal dichalcogenides (TMDs) have attracted considerable attention as atomically thin semiconductors for the ultimate transistor scaling. For practical applications in integrated electronics, large monolayer single crystals are essential for ensuring consistent electronic properties and high device yield. The TMDs available today are generally obtained by mechanical exfoliation or chemical vapor deposition (CVD) growth, but are often of mixed layer thickness, limited single crystal domain size or have very slow growth rate. Scalable and rapid growth of large single crystals of monolayer TMDs requires maximization of lateral growth rate while completely suppressing the vertical growth, which represents a fundamental synthetic challenge and has motivated considerable efforts. Herein we report a modified CVD approach with controllable reverse flow for rapid growth of large domain single crystals of monolayer TMDs. With the use of reverse flow to precisely control the chemical vapor supply in the thermal CVD process, we can effectively prevent undesired nucleation before reaching optimum growth temperature and enable rapid nucleation and growth of monolayer TMD single crystals at a high temperature that is difficult to attain with use of a typical thermal CVD process. We show that monolayer single crystals of 450 μm lateral size can be prepared in 10 s, with the highest lateral growth rate up to 45 μm/s. Electronic characterization shows that the resulting monolayer WSe2 material exhibits excellent electronic properties with carrier mobility up to 90 cm2 V−1 s−1, comparable to that of the best exfoliated monolayers. Our study provides a robust pathway for rapid growth of high-quality TMD single crystals. 相似文献
8.
In this paper, 3D particle focusing in a straight channel with asymmetrical expansion–contraction cavity arrays (ECCA channel) is achieved by exploiting the dean-flow-coupled elasto-inertial effects. First, the mechanism of particle focusing in both Newtonian and non-Newtonian fluids was introduced. Then particle focusing was demonstrated experimentally in this channel with Newtonian and non-Newtonian fluids using three different sized particles (3.2 μm, 4.8 μm, and 13 μm), respectively. Also, the effects of dean flow (or secondary flow) induced by expansion–contraction cavity arrays were highlighted by comparing the particle distributions in a single straight rectangular channel with that in the ECCA channel. Finally, the influences of flow rates and distances from the inlet on focusing performance in the ECCA channel were studied. The results show that in the ECCA channel particles are focused on the cavity side in Newtonian fluid due to the synthesis effects of inertial and dean-drag force, whereas the particles are focused on the opposite cavity side in non-Newtonian fluid due to the addition of viscoelastic force. Compared with the focusing performance in Newtonian fluid, the particles are more easily and better focused in non-Newtonian fluid. Besides, the Dean flow in visco-elastic fluid in the ECCA channel improves the particle focusing performance compared with that in a straight channel. A further advantage is three-dimensional (3D) particle focusing that in non-Newtonian fluid is realized according to the lateral side view of the channel while only two-dimensional (2D) particle focusing can be achieved in Newtonian fluid. Conclusively, this novel Dean-flow-coupled elasto-inertial microfluidic device could offer a continuous, sheathless, and high throughput (>10 000 s−1) 3D focusing performance, which may be valuable in various applications from high speed flow cytometry to cell counting, sorting, and analysis. 相似文献
9.
Jennifer S. Hartley M. Myintzu Hlaing Gediminas Seniutinas Saulius Juodkazis Paul R. Stoddart 《Biomicrofluidics》2015,9(6)
Surface-enhanced Raman scattering (SERS) shows promise for identifying single bacteria, but the short range nature of the effect makes it most sensitive to the cell membrane, which provides limited information for species-level identification. Here, we show that a substrate based on black silicon can be used to impale bacteria on nanoscale SERS-active spikes, thereby producing spectra that convey information about the internal composition of the bacterial capsule. This approach holds great potential for the development of microfluidic devices for the removal and identification of single bacteria in important clinical diagnostics and environmental monitoring applications.Plasma etching of silicon can be used to produce inexpensive, large surface area, nano-textured surfaces known as black silicon. Recently, it has been shown that black silicon nano-needles can impale bacteria1 and that it can be used as a sensor in microfluidic devices.2 When coated by a plasmonic metal, such as gold, the nano-textured surface of black silicon is ideal for use as a surface-enhanced Raman scattering (SERS) sensing platform.3 This work aims to investigate whether gold-coated black silicon nano-needles can be used to both impale bacteria and identify them by SERS. This combination of properties would promote the development of microfluidic devices for the removal and monitoring of bacteria in a wide range of medical, environmental, and industrial applications.4Black silicon was fabricated by a reactive ion etching technique,5 resulting in pyramidal-shaped spikes of height 185 ± 30 nm, full width at half height of 54 ± 10 nm, and 10 ± 2.4 nm radius of curvature at the tip. Samples were then magnetron sputter coated with 200 nm of gold, as this coating thickness was found to provide a suitable compromise between SERS enhancement and impalement efficiency. E. coli (ATCC 25922) from −80 °C stock was isolated on a nutrient agar plate (Difco nutrient broth, Becton Dickinson) for approximately 12 h. A single E. coli colony was then inoculated from the plate into 20 ml of nutrient broth media and incubated overnight at 37 °C with orbital shaking at 200 rpm. The total biomass of overnight culture was adjusted to an optical density of 0.3 at λ = 600 nm by adding fresh sterile nutrient broth (Cary 50 spectrophotometer, Agilent). The E. coli planktonic cells were washed three times by centrifugation at 12 000 rpm (Centrifuge 5804 R, Eppendorf) for 2 min. The washed cells were then re-suspended in a low strength minimum medium (Dulbecco A, phosphate buffered saline). A volume of 100 μl of solution was pipetted onto substrates and left to incubate for 1 h on the bench. Separate sets of samples were created for scanning electron microscope (SEM) imaging, live/dead staining, and SERS. Three sets were needed as each of these measurements altered the samples and left them unsuitable for further analysis.The first set of samples was washed three times with milliQ water after incubation, allowed to dry and then immediately sputter coated with gold using the Emitech K975x (operating current 35 mA, sputter time 32 s, stage rotation 138 rpm, and vacuum of 1 × 10−2 mbar). SEM imaging was performed with a Zeiss Supra 40VP in high vacuum mode with a working distance of 5 mm and an accelerating voltage of 3 kV. Figure Figure11 shows an example of the different levels of impalement that occurred on the black silicon surface. All cells showed signs of damage, but in some cases, the damage was limited to the perimeter of the cell and the main body appeared whole. In other cases, the entire cell had collapsed onto the spikes.Open in a separate windowFIG. 1.A typical SEM image showing E. coli cells with different levels of impalement on gold-coated black silicon.The second set of samples was used for live/dead staining (Invitrogen BacLight Bacterial Viability Kit L7012) with 3.34 mM SYTO 9 (green fluorescence) and 20 mM propidium iodide (red fluorescence). Equal volumes of both dyes were mixed thoroughly in a tube and added to the sample in a ratio of 3 μl of mixed dye to 1 ml of bacterial suspension. After mixing, a volume of 100 μl of the solution was pipetted onto the substrates, which were then incubated at room temperature in the dark for 15 min, before the staining solution was removed by pipetting. The substrates were then washed three times with milliQ water and mounted on a microscope slide for fluorescence imaging. The substrates were not allowed to dry and were stored in phosphate buffered saline at 4 °C when not in use. An epifluorescence microscope (Olympus IX71) with a mercury lamp source and a 60× water immersion objective was used to collect live/dead images from the substrates. Two filter blocks were used to collect the images: U-MNIBA2 blue excitation narrow band delivered green emission (live) and U-MWIG2 green excitation wide band provided red emission (dead).The live/dead image in Figure Figure22 shows a mix of both live and dead cells on the black silicon sample. The prevalence of live cells could be due to the incomplete impalement seen under SEM for some cells. It can also be explained by the sample still being wet during live/dead staining. The cells are dried prior to imaging in the SEM and this could weaken the cell wall and allow capillary forces to draw the cells onto the spikes for impalement. This hypothesis is supported by the large number of cells on the stained sample and the presence of cell groupings and cells imaged during mid-division. If the cells were immediately impaled, then such activity would not have been visible and a greater proportion of red cells would be expected.Open in a separate windowFIG. 2.Epifluorescence image showing live (green) and dead (red) E. coli cells after incubation on gold-coated black silicon.The third set of samples was washed three times with milliQ water after incubation and allowed to dry prior to spectral analysis. SERS spectra were collected with a Renishaw inVia Raman spectrometer operating at 785 nm with a 1200 l/mm grating. Power at the sample was 150 mW focused with a 100 × /0.85 NA objective to obtain a diffraction limited laser spot. The resulting spot size (≤2 μm in diameter) is well matched to the size of the bacterial cells. Spectra were collected with three accumulations of 10 s. Data were background subtracted6 and normalised to unity for ease of plotting. A great deal of variability was observed in the resulting spectra, as shown in Figure Figure33.Open in a separate windowFIG. 3.SERS spectra of E. coli after incubation on a gold-coated black silicon substrate. The spectrum numbers represent single cells at different locations and different levels of impalement.It should be noted that E. coli SERS is known to produce a high level of variability,7–12 depending on the experimental setup.13 However, the variability seen in the SERS spectra of Fig. Fig.33 is unusual for measurements performed under consistent experimental conditions. This increased level of variability may be related to the different levels of impalement seen in Fig. Fig.1,1, which results in the probing of different internal components. SERS is a surface sensitive technique, with the signal primarily arising within 2 nm of the metal surface.14 Note that unlike apertureless nanoprobes15 or conical plasmonic nanotips,16 the SERS signal in black silicon arises primarily from “hot spots” between the spikes, where the plasmon resonance field is particularly strong.17 Therefore, depending on the depth and location of impalement, different biomolecules are expected to be excited by this novel substrate.Some peaks occur in the same position for multiple spectra (e.g., peak positions 420, 893, 1001, 1285, and 1307 cm−1), but there are also a lot of unique peaks. The vertical lines in Fig. Fig.33 indicate peaks which have appeared in the literature for SERS of E. coli.7–12 Spectrum 3 has a high proportion of peaks matching published values. This is also the case for spectrum 5, which shares a few peak positions with spectrum 3. Preliminary peak allocations have identified carbohydrates11 (420 cm−1), tyrosine11 (650 cm−1), adenine10,11 (706 and 735 cm−1), hypoxanthine7 (722 and1373 cm−1), phenylalanine9 (1001 cm−1), amide III (Ref. 10) (1285 cm−1), CH2 deformation12 (1556 cm−1), and C=C10 (1587 cm−1).Given the varying levels of impalement observed in the SEM, it appears that the spike shape and Au coating should be further optimized to ensure that the entire cell is consistently pierced and the internal biomolecules are more comprehensively probed. In this way, it may be possible to obtain a more reproducible SERS spectrum of the internal biomolecular constituents of single bacterial cells, thereby providing rapid identification for medical and environmental diagnostic applications. Given that SERS is insensitive to water,4 future work should aim to achieve impalement in an aqueous environment, so that the full capability of microfluidics can be used to separate and concentrate suspended bacteria before presenting them to the substrate for rapid analysis.4 This suggests a broad range of potential applications in the detection, monitoring, and control of bacterial contamination. 相似文献
10.
Aarash Sofla Bojana Cirkovic Anne Hsieh Jason W. Miklas Nenad Filipovic Milica Radisic 《Biomicrofluidics》2013,7(1)
The majority of available cardiomyocyte markers are intercellular proteins, limiting our ability to enrich live cardiomyocytes from heterogeneous cell preparations in the absence of genetic labeling. Here, we describe enrichment of live cardiomyocytes from the hearts of adult mice in a label-free microfluidic approach. The separation device consisted of a vertical column (15 mm long, 700 μm diameter), placed between permanent magnets resulting in a field strength of 1.23 T. To concentrate the field at the column wall, the column was wrapped with 69 μm diameter nickel wire. Before passing the cells through the column, the cardiomyocytes in the cell suspension had been rendered paramagnetic by treatment of the adult mouse heart cell preparation with sodium nitrite (2.5 mM) for 20 min on ice. The cell suspension was loaded into the vertical column from the top and upon settling, the non-myocytes were removed by the upward flow from the column. The cardiomyocytes were then collected from the column by applying a higher flow rate (144 μl/min). We found that by applying a separation flow rate of 4.2 μl/min in the first step, we can enrich live adult cardiomyocytes to 93% ± 2% in a label-free manner. The cardiomyocytes maintained viability immediately after separation and upon 24 h in culture. 相似文献
11.
Recent years have witnessed a strong trend towards analysis of single-cells. To access and handle single-cells, many new tools are needed and have partly been developed. Here, we present an improved version of a single-cell printer which is able to deliver individual single cells and beads encapsulated in free-flying picoliter droplets at a single-bead efficiency of 96% and with a throughput of more than 10 beads per minute. By integration of acoustophoretic focusing, the cells could be focused in x and y direction. This way, the cells were lined-up in front of a 40 μm nozzle, where they were analyzed individually by an optical system prior to printing. In agreement with acoustic simulations, the focusing of 10 μm beads and Raji cells has been achieved with an efficiency of 99% (beads) and 86% (Raji cells) to a 40 μm wide center region in the 1 mm wide microfluidic channel. This enabled improved optical analysis and reduced bead losses. The loss of beads that ended up in the waste (because printing them as single beads arrangements could not be ensured) was reduced from 52% ± 6% to 28% ± 1%. The piezoelectric transducer employed for cell focusing could be positioned on an outer part of the device, which proves the acoustophoretic focusing to be versatile and adaptable. 相似文献
12.
This article describes a fabrication process for the generation of a leak proof paper based microfluidic device and a new design strategy for convenient incorporation of externally prepared test zones. Briefly, a negative photolithographic method was used to prepare the device with a partial photoresist layer on the rear of the device to block the leakage of sample. Microscopy and Field Emission Scanning Electron Microscopy data validated the formation of the photoresist layer. The partial layer of photoresist on the device channel limits sample volume to 7 ± 0.2 μl as compared to devices without the partial photoresist layer which requires a larger sample volume of 10 ± 0.1 μl. The design prototype with a customized external test zone exploits the channel protrusions on the UV exposed photoresist treated paper to bridge the externally applied test zone to the sample and absorbent zones. The partially laminated device with an external test zone has a comparatively low wicking speed of 1.8 ± 0.9 mm/min compared to the completely laminated device with an inbuilt test zone (3.3 ± 1.2 mm/min) which extends the reaction time between the analyte and reagents. The efficacy of the prepared device was studied with colorimetric assays for the non-specific detection of protein by tetrabromophenol blue, acid/base with phenolphthalein indicator, and specific detection of proteins using the HRP-DAB chemistry. The prepared device has the potential for leak proof detection of analyte, requires low sample volume, involves reduced cost of production (∼$0.03, excluding reagent and lamination cost), and enables the integration of customized test zones. 相似文献
13.
Jyotsna A. Patil Mandakini S. Kshirsagar Arun J. Patil 《Indian journal of clinical biochemistry : IJCB》2021,36(1):94
Activated carbon fabrics (ACF) mask prevents the absorption of lead and reduce its adverse effects of human health. Aim of this study to know the blood lead level and its effects on heme biosynthesis and hematological parameters after using 2 months activated carbon fabric mask of battery manufacturing workers (BMW). Blood lead level, heme biosynthesis and hematological parameters were measured by using standard method. Blood lead level (P < 0.001, − 13.5%) was significantly decreased, activated δ-aminolevulinic acid dehydratase (P < 0.001, 11.97%) and non-activated δ- aminolevulinic acid dehydratase (P < 0.001, 23.17%) enzyme activity were significantly increased, however, the ratio of activated to Non-activated δ- ALAD (P < 0.001, − 10.13%) was significantly decreased, urinary excretion of δ- aminolevulinic acid (P < 0.001, − 10.49%) and porphobilinogen (P < 0.001, − 7.38%) were significantly decreased after using 2 months ACF mask as compared to before using mask of BMW. Hematological parameters i.e Hb (P < 0.05, 13.42%), PCV (P < 0.05, 7.23%), MCV (P < 0.05, 1.9%) were significantly increased and total WBC count (P < 0.05, − 5.18%) was significantly decreased after using 2 months ACF mask as compared to before using mask of BMW. Two months using ACF mask reduces the blood lead level and improves the δ-ALDH activity and hematological parameters, decreases the urinary excretion of δ-ALA, PBG of battery manufacturing workers. Therefore, the regular using of ACF mask is beneficial to prevent the lead absorption and its adverse effects on human health. 相似文献
14.
Applying metal organic frameworks (MOFs) in electrochemical systems is a currently emerging field owing to the rich metal nodes and highly specific surface area of MOFs. However, the problems for MOFs that need to be solved urgently are poor electrical conductivity and low ion transport. Here we present a facile in situ growth method for the rational synthesis of MOFs@hollow mesoporous carbon spheres (HMCS) yolk–shell-structured hybrid material for the first time. The size of the encapsulated Zeolitic Imidazolate Framework-67 (ZIF-67) is well controlled to 100 nm due to the spatial confinement effect of HMCS, and the electrical conductivity of ZIF-67 is also increased significantly. The ZIF@HMCS-25% hybrid material obtained exhibits a highly efficient oxygen reduction reaction activity with 0.823 V (vs. reversible hydrogen electrode) half-wave potential and an even higher kinetic current density (JK = 13.8 mA cm−2) than commercial Pt/C. ZIF@HMCS-25% also displays excellent oxygen evolution reaction performance and the overpotential of ZIF@HMCS-25% at 10 mA cm−2 is 407 mV. In addition, ZIF@HMCS-25% is further employed as an air electrode for a rechargeable Zn–air battery, exhibiting a high power density (120.2 mW cm−2 at 171.4 mA cm−2) and long-term charge/discharge stability (80 h at 5 mA cm−2). This MOFs@HMCS yolk–shell design provides a versatile method for the application of MOFs as electrocatalysts directly. 相似文献
15.
Shimul C. Saha Andrew M. Powl B. A. Wallace Maurits R. R. de Planque Hywel Morgan 《Biomicrofluidics》2015,9(1)
We describe a scalable artificial bilayer lipid membrane platform for rapid electrophysiological screening of ion channels and transporters. A passive pumping method is used to flow microliter volumes of ligand solution across a suspended bilayer within a microfluidic chip. Bilayers are stable at flow rates up to ∼0.5 μl/min. Phospholipid bilayers are formed across a photolithographically defined aperture made in a dry film resist within the microfluidic chip. Bilayers are stable for many days and the low shunt capacitance of the thin film support gives low-noise high-quality single ion channel recording. Dose-dependent transient blocking of α-hemolysin with β-cyclodextrin (β-CD) and polyethylene glycol is demonstrated and dose-dependent blocking studies of the KcsA potassium channel with tetraethylammonium show the potential for determining IC50 values. The assays are fast (30 min for a complete IC50 curve) and simple and require very small amounts of compounds (100 μg in 15 μl). The technology can be scaled so that multiple bilayers can be addressed, providing a screening platform for ion channels, transporters, and nanopores. 相似文献
16.
Rui Zhang Jie Huang Fei Xie Baojun Wang Ming Chu Yuedan Wang Haichao Li Wei Wang Haixia Zhang Wengang Wu Zhihong Li 《Biomicrofluidics》2014,8(3)
Nowadays, microfluidics is attracting more and more attentions in the biological society and has
provided powerful solutions for various applications. This paper reported a microfluidic strategy
for aqueous sample sterilization. A well-designed small microchannel with a high hydrodynamic
resistance was used to function as an in-chip pressure regulator. The pressure in the upstream
microchannel was thereby elevated which made it possible to maintain a boiling-free high temperature
environment for aqueous sample sterilization. A 120 °C temperature along with a pressure of 400 kPa
was successfully achieved inside the chip to sterilize aqueous samples with E. coli
and Staphylococcus aureus inside. This technique will find wide applications in
portable cell culturing, microsurgery in wild fields, and other related micro total analysis
systems.Microfluidics, which confines fluid flow at microscale, attracts more and more attentions in the
biological society.1–4 By scaling the flow
domain down to microliter level, microfluidics shows attractive merits of low sample consumption,
precise biological objective manipulation, and fast momentum/energy transportation. For example,
various cell operations, such as culturing5–7
and sorting,8–10 have already been
demonstrated with microfluidic approaches. In most biological applications, sterilization is a key
sample pre-treatment step to avoid contamination. However, as far as the author knew, this important
pre-treatment operation is generally achieved in an off-chip way, by using high temperature and high
pressure autoclave. Actually, microfluidics has already been utilized to develop new solution for
high pressure/temperature reactions. The required high pressure/temperature condition was generated
either by combining off-chip back pressure regulator and hot-oil bath,11,12 or by integrating pressure regulator, heater, and temperature
sensor into a single chip.13 This work presented a
microfluidic sterilization strategy by implementing the previously developed continuous flowing high
pressure/temperature microfluidic reactor.Figure Figure11 shows the working principle of the present
microfluidic sterilization chip. The chip consists of three zones: sample loading (a microchannel
with length of 270 mm and width of 40 μm), sterilization (length of 216 mm and
width of 100 μm), and pressure regulating (length of 42 mm and width of
5 μm). Three functional zones were separated by two thermal isolation trenches. The
sample was injected into the chip by a syringe pump and experienced two-step filtrations (feature
sizes of 20 μm and 5 μm, not shown in Figure Figure1)1) at the entrance to avoid the channel clog. All channels had the same depth of
40 μm. According to the Hagen–Poiseuille relationship,15 the pressure regulating channel had a large flow resistance (around
1.09 × 1017 Pa·s/m3, see supplementary S1 for details16) because of its small width, thereby generated a high working
pressure in the upstream sterilization channel under a given flow rate. The boiling point of the
solution will then be raised up by the elevated pressure in the sterilization zone followed by the
Antoine equation.16 By integrating
heater/temperature sensors in the pressurized zone, a high temperature environment with temperature
higher than 100 °C can thereby be realized for aqueous sample sterilization. The sample was
collected from the outlet and cultured at 37 °C for 12 h. Bacterial colony was counted to evaluate
the sterilization performance.Open in a separate windowFIG. 1.Working principle of the present microfluidic sterilization. Only microfluidic channel, heater,
and temperature sensor were schematically shown. The varied colour of the microchannel represents
the pressure and that of the halation stands for the temperature.Fabrication of this chip has been introduced elsewhere.14 The fabricated chip and the experimental system are shown in Figure Figure2.2. There were two inlets of the chip. While, in the experiment, only
one inlet used and connected to the syringe pump. The backup one was blocked manually. The sample
load zone was arranged in between of the sterilization zone and the pressure regulating zone based
on thermal management consideration. A temperature control system (heater/temperature sensor, power
source, and multi-meter) was setup to provide the required high temperature. The heater and the
temperature sensor were microfabricated Pt resistors. The temperature coefficient of resistance
(TCR) was measured as 0.00152 K−1.Open in a separate windowFIG. 2.The fabricated chip and the experimental system. (a) Two chips with a penny for comparison. The
left chip was viewed from the heater/temperature sensor side, while the right one was observed from
the microchannel side (through a glass substrate). (b) The experimental system.Thermal isolation performance of the present chip before packaging with inlet/outlet was shown in
Figure Figure3,3, to show the thermal interference issue. The results
indicated that when the sterilization zone was heated up to 140 °C, the pressure regulating zone was
about 40 °C. At this temperature, the viscosity of water decreases to 0.653 mPa·s from 1.00 mPa·s
(at 20 °C), which will make the pressure in the sterilization zone reduced from 539 kPa (calculated
at 20 °C and flow rate of 4 nl/s) to 387 kPa. The boiling point will then decrease to 142.8 °C,
which will guarantee a boiling-free sterilization. In the cases without the thermal isolation
trenches, the temperature of the pressure regulating zone reached as high as 75 °C because of the
thermal interference from the sterilization zone, as shown in Figure Figure3.3. The pressure in the sterilization zone was then reduced to 268 kPa (calculated at flow
rate of 4 nl/s) and the boiling temperature was around 130 °C, which was lower than the set
sterilization temperature. Detail calculation can be found in supplementary S2.16Open in a separate windowFIG. 3.The temperature distribution of the chips (before packaged) with and without thermal isolation
trenches (powered at 1 W). The data were extracted from the central lines of infrared images, as
shown as inserts.Bacterial sterilization performance of the present chip was tested and the experimental results
were shown in Figure Figure4.4. E. coli with initial
concentration of 106/ml was pumped into and flew through the chip with the sterilization
temperatures varied from 25 °C to 120 °C at flow rates of 2 nl/s and 4 nl/s. The outflow was
collected and inoculated onto the SS agar plate evenly with inoculation loops. The population of
bacteria in the outflow was counted based on the bacterial colonies after incubation at 37 °C for
12 h. Typical bacterial colonies were shown in Figure Figure4.4. The
low flow rate case showed a better sterilization performance because of the longer staying period in
the sterilization channel. The population of E. coli was around
1.25 × 104/ml after a 432 s-long, 70 °C sterilization (at flow rate of 2 nl/s). While at
the flow rate of 4 nl/s, the cultivation result indicated the population was around
3.8 × 104/ml because the sterilization time was shorten to 216 s. A control case, where
the solution flew through an un-heated chip at 2 nl/s, was conducted to investigate the effect of
the shear stress on the sterilization performance (see the supplementary S3 for details16). As listed in Table TableI,I, the results indicated that the shear stress did not show any noticeable effect on the
bacterial sterilization. When the chip was not heated, i.e., the case with the largest shear stress
because of the highest viscosity of fluid, the bacterial cultivation was nearly the same as the
off-chip results (no stress). The temperature has the most significant effect on the sterilization
performance. No noticeable bacteria proliferation was observed in the cases with the sterilization
temperature higher than 100 °C, as shown in Figure Figure44.
Open in a separate windowaData in the table are shear stress (Pa)/population of bacteria, where “+++” indicates a large
proliferation, “+” means small but noticeable proliferation, “−” represents no proliferation.bOff-chip control group.Open in a separate windowFIG. 4.Sterilization performance of the present chip with E. coli and S.
aureus as test bacteria. All the original population was 106/ml. Inserted images
showed the images of the culture disk after bacteria incubation.Sterilization of another commonly encountered bacterium, Staphylococcus aureus,
with initial population of 106/ml was also tested in the present chip, as shown in Figure
Figure4.4. Similarly, no noticeable S. aureus
proliferation was found when the sterilization temperature was higher than 100 °C.In short, we demonstrated a microfluidic sterilization strategy by utilizing a continuous flowing
high temperature/pressure chip. The population of E. coli or S.
aureus was reduced from 106/ml to an undetectable level when the sterilization
temperature of the chip was higher than 100 °C. The chip holds promising potential in developing
portable microsystem for biological/clinical applications. 相似文献
Table I.
The E. coli cultivation results under different flow rates and sterilization temperatures. a| 25 °C | 70 °C | 100 °C | 120 °C | 25 °C b | |
|---|---|---|---|---|---|
| 2 nl/s | 1.89/+++ | 1.38/+ | 1.16/− | 1.04/− | 0/+++ |
| 4 nl/s | 3.78/+++ | 2.76/+ | 2.32/− | 2.08/− | 0/+++ |
17.
Orloff ND Dennis JR Cecchini M Schonbrun E Rocas E Wang Y Novotny D Simmonds RW Moreland J Takeuchi I Booth JC 《Biomicrofluidics》2011,5(4):44107-441079
We present a 91 MHz surface acoustic wave resonator with integrated microfluidics that includes a flow focus, an expansion region, and a binning region in order to manipulate particle trajectories. We demonstrate the ability to change the position of the acoustic nodes by varying the electronic phase of one of the transducers relative to the other in a pseudo-static manner. The measurements were performed at room temperature with 3 μm diameter latex beads dispersed in a water-based solution. We demonstrate the dependence of nodal position on pseudo-static phase and show simultaneous control of 9 bead streams with spatial control of −0.058 μm/deg ± 0.001 μm/deg. As a consequence of changing the position of bead streams perpendicular to their flow direction, we also show that the integrated acoustic-microfluidic device can be used to change the trajectory of a bead stream towards a selected bin with an angular control of 0.008 deg/deg ± 0.000(2) deg/deg. 相似文献
18.
Cong Liu Feng An Paria S M Gharavi Qinwen Lu Junkun Zha Chao Chen Liming Wang Xiaozhi Zhan Zedong Xu Yuan Zhang Ke Qu Junxiang Yao Yun Ou Zhiming Zhao Xiangli Zhong Dongwen Zhang Nagarajan Valanoor Lang Chen Tao Zhu Deyang Chen Xiaofang Zhai Peng Gao Tingting Jia Shuhong Xie Gaokuo Zhong Jiangyu Li 《国家科学评论(英文版)》2020,7(1):84
Complex oxides with tunable structures have many fascinating properties, though high-quality complex oxide epitaxy with precisely controlled composition is still out of reach. Here we have successfully developed solution-based single-crystalline epitaxy for multiferroic (1-x)BiTi(1-y)/2FeyMg(1-y)/2O3–(x)CaTiO3 (BTFM–CTO) solid solution in large area, confirming its ferroelectricity at the atomic scale with strong spontaneous polarization. Careful compositional tuning leads to a bulk magnetization of 0.07 ± 0.035 μB/Fe at room temperature, enabling magnetically induced polarization switching exhibiting a large magnetoelectric coefficient of 2.7–3.0 × 10−7 s/m. This work demonstrates the great potential of solution processing in large-scale complex oxide epitaxy and establishes novel room-temperature magnetoelectric coupling in epitaxial BTFM–CTO film, making it possible to explore a much wider space of composition, phase, and structure that can be easily scaled up for industrial applications. 相似文献
19.
Genetic sequence and hyper-methylation profile information from the promoter regions of tumor suppressor genes are important for cancer disease investigation. Since hyper-methylated DNA (hm-DNA) is typically present in ultra-low concentrations in biological samples, such as stool, urine, and saliva, sample enrichment and amplification is typically required before detection. We present a rapid microfluidic solid phase extraction (μSPE) system for the capture and elution of low concentrations of hm-DNA (≤1 ng ml−1), based on a protein-DNA capture surface, into small volumes using a passive microfluidic lab-on-a-chip platform. All assay steps have been qualitatively characterized using a real-time surface plasmon resonance (SPR) biosensor, and quantitatively characterized using fluorescence spectroscopy. The hm-DNA capture/elution process requires less than 5 min with an efficiency of 71% using a 25 μl elution volume and 92% efficiency using a 100 μl elution volume. 相似文献
20.
Blood analysis plays a major role in medical and science applications and white blood cells (WBCs) are an important target of analysis. We proposed an integrated microfluidic chip for direct and rapid trapping WBCs from whole blood. The microfluidic chip consists of two basic functional units: a winding channel to mix and arrays of two-layer trapping structures to trap WBCs. Red blood cells (RBCs) were eliminated through moving the winding channel and then WBCs were trapped by the arrays of trapping structures. We fabricated the PDMS (polydimethylsiloxane) chip using soft lithography and determined the critical flow velocities of tartrazine and brilliant blue water mixing and whole blood and red blood cell lysis buffer mixing in the winding channel. They are 0.25 μl/min and 0.05 μl/min, respectively. The critical flow velocity of the whole blood and red blood cell lysis buffer is lower due to larger volume of the RBCs and higher kinematic viscosity of the whole blood. The time taken for complete lysis of whole blood was about 85 s under the flow velocity 0.05 μl/min. The RBCs were lysed completely by mixing and the WBCs were trapped by the trapping structures. The chip trapped about 2.0 × 103 from 3.3 × 103 WBCs. 相似文献