Miniaturization of dielectric liquid microlens in package

This study presents packaged microscale liquid lenses actuated with liquid droplets of 300-700 μm in diameter using the dielectric force manipulation. The liquid microlens demonstrated function focal length tunability in a plastic package. The focal length of the liquid lens with a lens droplet of 500 μm in diameter is shortened from 4.4 to 2.2 mm when voltages applied change from 0 to 79 V(rms). Dynamic responses that are analyzed using 2000 frames∕s high speed motion cameras show that the advancing and receding times are measured to be 90 and 60 ms, respectively. The size effect of dielectric liquid microlens is characterized for a lens droplet of 300-700 μm in diameter in an aspect of focal length.     

Observation of nonspherical particle behaviors for continuous shape-based separation using hydrodynamic filtration

Selection of particles or cells of specific shapes from a complex mixture is an essential procedure for various biological and industrial applications, including synchronization of the cell cycle, classification of environmental bacteria, and elimination of aggregates from synthesized particles. Here, we investigate the separation behaviors of nonspherical and spherical particles∕cells in the hydrodynamic filtration (HDF) scheme, which was previously developed for continuous size-dependent particle∕cell separation. Nonspherical particle models were prepared by coating the hemisphere of spherical polymer particles with a thin Au layer and by bonding the Janus particles to form twins and triplets resembling dividing and aggregating cells, respectively. High-speed imaging revealed a difference in the separation behaviors of spherical and nonspherical particles at a branch point; nonspherical particles showed rotation behavior and did not enter the branch channel even when their minor axis was smaller than the virtual width of the flow region entering the branch channel, w(1). The confocal-laser high-speed particle intensity velocimetry system visualized the flow profile inside the HDF microchannel, demonstrating that the steep flow-velocity distribution at the branch point is the main factor causing the rotation behavior of nonspherical particles. As applications, we successfully separated spherical and nonspherical particles with various major∕minor lengths and also demonstrated the selection of budding∕single cells from a yeast cell mixture. We therefore conclude that the HDF scheme can be used for continuous shape-based particle∕cell separation.     

High-throughput size-based rare cell enrichment using microscale vortices

Cell isolation in designated regions or from heterogeneous samples is often required for many microfluidic cell-based assays. However, current techniques have either limited throughput or are incapable of viable off-chip collection. We present an innovative approach, allowing high-throughput and label-free cell isolation and enrichment from heterogeneous solution using cell size as a biomarker. The approach utilizes the irreversible migration of particles into microscale vortices, developed in parallel expansion-contraction trapping reservoirs, as the cell isolation mechanism. We empirically determined the critical particle∕cell diameter D(crt) and the operational flow rate above which trapping of cells∕particles in microvortices is initiated. Using this approach we successfully separated larger cancer cells spiked in blood from the smaller blood cells with processing rates as high as 7.5×10(6) cells∕s. Viable long-term culture was established using cells collected off-chip, suggesting that the proposed technique would be useful for clinical and research applications in which in vitro culture is often desired. The presented technology improves on current technology by enriching cells based on size without clogging mechanical filters, employing only a simple single-layered microfluidic device and processing cell solutions at the ml∕min scale.     

Passive optical separation and enrichment of cells by size difference

A size-selective cell sorting microfluidic device that utilizes optical force is developed. The device consists of a three-dimensional polydimethylsiloxane microstructure comprised of two crossed microchannels in a three-dimensional configuration. A line shaped focused laser beam is used for automatic size-selective cell sorting in a continuous flow environment. As yeast cells in an aqueous medium are fed continuously into a lower channel, the line shaped focused laser beam is applied (perpendicular to the direction of flow) at the junction of the two crossed channels. The scattering force of the laser beam was employed to push cells matching specific criteria upward from one channel to another. The force depends on the size of the cells, the laser power, and the fluid flow speed. The variation in size of yeast cells causes them to follow different routes at the intersection. For flow speeds below 30 μm∕s, all yeast cells larger than 3 μm were removed from the main stream. As a result, a high purity sample of small cells can be collected at the outlet of bottom channel.     

Transmittance tuning by particle chain polarization in electrowetting-driven droplets

A tiny droplet containing nano∕microparticles commonly handled in digital microfluidic lab-on-a-chip is regarded as a micro-optical component with tunable transmittance at programmable positions for the application of micro-opto-fluidic-systems. Cross-scale electric manipulations of droplets on a millimeter scale as well as suspended particles on a micrometer scale are demonstrated by electrowetting-on-dielectric (EWOD) and particle chain polarization, respectively. By applying electric fields at proper frequency ranges, EWOD and polarization can be selectively achieved in designed and fabricated parallel plate devices. At low frequencies, the applied signal generates EWOD to pump suspension droplets. The evenly dispersed particles reflect and∕or absorb the incident light to exhibit a reflective or dark droplet. When sufficiently high frequencies are used on to the nonsegmented parallel electrodes, a uniform electric field is established across the liquid to polarize the dispersed neutral particles. The induced dipole moments attract the particles each other to form particle chains and increase the transmittance of the suspension, demonstrating a transmissive or bright droplet. In addition, the reflectance of the droplet is measured at various frequencies with different amplitudes.     

dc electrokinetic transport of cylindrical cells in straight microchannels

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

Curvature-induced dielectrophoresis for continuous separation of particles by charge in spiral microchannels

The separation of particles from a heterogeneous mixture is critical in chemical and biological analyses. Many methods have been developed to separate particles in microfluidic devices. However, the majority of these separations have been limited to be size based and binary. We demonstrate herein a continuous dc electric field driven separation of carboxyl-coated and noncoated 10 μm polystyrene beads by charge in a double-spiral microchannel. This method exploits the inherent electric field gradients formed within the channel turns to manipulate particles by dielectrophoresis and is thus termed curvature-induced dielectrophoresis. The spiral microchannel is also demonstrated to continuously sort noncoated 5 μm beads, noncoated 10 μm beads, and carboxyl-coated 10 μm beads into different collecting wells by charge and size simultaneously. The observed particle separation processes in different situations are all predicted with reasonable agreements by a numerical model. This curvature-induced dielectrophoresis technique eliminates the in-channel microelectrodes and obstacles that are required in traditional electrode- and insulator-based dielectrophoresis devices. It may potentially be used to separate multiple particle targets by intrinsic properties for lab-on-a-chip applications.     

Electrochemical biosensors for on-chip detection of oxidative stress from immune cells

Seamless integration of biological components with electrochemical sensors is critical in the development of microdevices for cell analysis. The present paper describes the integration miniature Au electrodes next to immune cells (macrophages) in order to detect cell-secreted hydrogen peroxide (H(2)O(2)). Photopatterning of poly(ethylene glycol) (PEG) hydrogels was used to both immobilize horseradish peroxidase molecules onto electrodes and to define regions for cell attachment in the vicinity of sensing electrodes. Electrodes micropatterned in such a manner were enclosed inside poly(dimethylsiloxane) fluid conduits and incubated with macrophages. The cells attached onto the exposed glass regions in the vicinity of the electrodes and nowhere else on the non-fouling PEG hydrogel surface. A microfluidic device was converted into an electrochemical cell by placing flow-through Ag∕AgCl reference and Pt wire counter electrodes at the outlet and inlet, respectively. This microdevice with integrated H(2)O(2)-sensing electrodes had sensitivity of 27 μA∕cm(2) mM with a limit of detection of 2 μM. Importantly, this microdevice allowed controllable seeding of macrophages next to electrodes, activation of these cells and on-chip monitoring of H(2)O(2) release in real time. In the future, this biosensor platform may be utilized for monitoring of macrophage responses to pathogens or for the study of inflammatory signaling in micropatterned cell cultures.     

Optofluidic planar reactors for photocatalytic water treatment using solar energy

Optofluidics may hold the key to greater success of photocatalytic water treatment. This is evidenced by our findings in this paper that the planar microfluidic reactor can overcome the limitations of mass transfer and photon transfer in the previous photocatalytic reactors and improve the photoreaction efficiency by more than 100 times. The microreactor has a planar chamber (5 cm×1.8 cm×100 μm) enclosed by two TiO(2)-coated glass slides as the top cover and bottom substrate and a microstructured UV-cured NOA81 layer as the sealant and flow input∕output. In experiment, the microreactor achieves 30% degradation of 3 ml 3×10(-5)M methylene blue within 5 min and shows a reaction rate constant two orders higher than the bulk reactor. Under optimized conditions, a reaction rate of 8% s(-1) is achieved under solar irradiation. The average apparent quantum efficiency is found to be only 0.25%, but the effective apparent quantum efficiency reaches as high as 25%. Optofluidic reactors inherit the merits of microfluidics, such as large surface∕volume ratio, easy flow control, and rapid fabrication and offer a promising prospect for large-volume photocatalytic water treatment.     

Polyelectrolyte multilayer surface functionalization of poly(dimethylsiloxane) (PDMS) for reduction of yeast cell adhesion in microfluidic devices

Polyelectrolyte multilayers (PEMs) based on the combinations poly(diallyldimethylammonium chloride)∕poly(acrylic acid) (PDADMAC∕PAA) and poly(allylamine hydrochloride)∕PAA (PAH∕PAA) were adsorbed on poly(dimethylsiloxane) (PDMS) and tested for nonspecific surface attachment of hydrophobic yeast cells using a parallel plate flow chamber. A custom-made graft copolymer containing poly(ethylene glycol) (PEG) side chains (PAA-g-PEG) was additionally adsorbed on the PEMs as a terminal layer. A suitable PEM modification effectively decreased the adhesion strength of Saccharomyces cerevisiae DSM 2155 to the channel walls. However, a further decrease in initial cell attachment and adhesion strength was observed after adsorption of PAA-g-PEG copolymer onto PEMs from aqueous solution. The results demonstrate that a facile layer-by-layer surface functionalization from aqueous solutions can be successfully applied to reduce cell adhesion strength of S. cerevisiae by at least two orders of magnitude compared to bare PDMS. Therefore, this method is potentially suitable to promote planktonic growth inside capped PDMS-based microfluidic devices if the PEM deposition is completed by a dynamic flow-through process.     

Resistive pulse sensing of magnetic beads and supraparticle structures using tunable pores

Tunable pores (TPs) have been used for resistive pulse sensing of 1 μm superparamagnetic beads, both dispersed and within a magnetic field. Upon application of this field, magnetic supraparticle structures (SPSs) were observed. Onset of aggregation was most effectively indicated by an increase in the mean event magnitude, with data collected using an automated thresholding method. Simulations enabled discrimination between resistive pulses caused by dimers and individual particles. Distinct but time-correlated peaks were often observed, suggesting that SPSs became separated in pressure-driven flow focused at the pore constriction. The distinct properties of magnetophoretic and pressure-driven transport mechanisms can explain variations in the event rate when particles move through an asymmetric pore in either direction, with or without a magnetic field applied. Use of TPs for resistive pulse sensing holds potential for efficient, versatile analysis and measurement of nano- and microparticles, while magnetic beads and particle aggregation play important roles in many prospective biosensing applications.     

Antibiotic susceptibility test based on the dielectrophoretic behavior of elongated Escherichia coli with cephalexin treatment

This study reports the use of dielectrophoresis (DEP), which determined the crossover frequency (cof) of antibiotic-induced elongation of Escherichia coli (E. coli) with regard to the rapid antibiotic susceptibility test (AST). Different dielectric properties and elongation rates of E. coli are caused by various concentrations of cephalexin treatment. According to the authors' results, significant changes in the cof of bacteria treated with 32 μg∕ml antibiotic for 60 min can be found by using a quadruple electrode array, and the results of DEP-based AST correspond with that of agar dilution method. Utilizing this approach could greatly reduce the period of bacteria growth, and obtain the minimum inhibition concentration of E. coli to cephalexin.     

Structural and dynamic properties of linker histone H1 binding to DNA

Found in all eukaryotic cells, linker histones H1 are known to bind to and rearrange nucleosomal linker DNA. In vitro, the fundamental nature of H1∕DNA interactions has attracted wide interest among research communities-from biologists to physicists. Hence, H1∕DNA binding processes and structural and dynamical information about these self-assemblies are of broad importance. Targeting a quantitative understanding of H1 induced DNA compaction mechanisms, our strategy is based on using small-angle x-ray microdiffraction in combination with microfluidics. The usage of microfluidic hydrodynamic focusing devices facilitates a microscale control of these self-assembly processes, which cannot be achieved using conventional bulk setups. In addition, the method enables time-resolved access to structure formation in situ, in particular, to transient intermediate states. The observed time dependent structure evolution shows that the H1∕DNA interaction can be described as a two-step process: an initial unspecific binding of H1 to DNA is followed by a rearrangement of molecules within the formed assemblies. The second step is most likely induced by interactions between the DNA and the H1's charged side chains. This leads to an increase in lattice spacing within the DNA∕protein assembly and induces a decrease in the correlation length of the mesophases, probably due to a local bending of the DNA.     

Characterization of a microflow cytometer with an integrated three-dimensional optofluidic lens system

Flow cytometry is a standard analytical method in cell biology and clinical diagnostics and is widely distributed for the experimental investigation of microparticle characteristics. In this work, the design, realization, and measurement results of a novel planar optofluidic flow cytometric device with an integrated three-dimensional (3D) adjustable optofluidic lens system for forward-scattering∕extinction-based biochemical analysis fabricated by silicon micromachining are presented. To our knowledge, this is the first planar cytometric system with the ability to focus light three-dimensionally on cells∕particles by the application of fluidic lenses. The single layer microfluidic platform enables versatile 3D hydrodynamic sample focusing to an arbitrary position in the channel and incorporates integrated fiber grooves for the insertion of glass fibers. To confirm the fluid dynamics and raytracing simulations and to characterize the sensor, different cell lines and sets of microparticles were investigated by detecting the extinction (axial light loss) signal, demonstrating the high sensitivity and sample discrimination capability of this analysis system. The unique features of this planar microdevice enable new biotechnological analysis techniques due to the highly increased sensitivity.     

Two-dimensional spatial manipulation of microparticles in continuous flows in acoustofluidic systems

We report a modeling and experimental study of techniques to acoustically focus particles flowing through a microfluidic channel. Our theoretical model differs from prior works in that we solve an approximate 2-D wave transmission model that accounts for wave propagation in both the solid and fluid phases. Our simulations indicate that particles can be effectively focused at driving frequencies as high as 10% off of the resonant condition. This conclusion is supported by experiments on the acoustic focusing of particles in nearly square microchannels, which are studied for different flow rates, driving frequencies and placements of the lead zirconate titanate transducer, either underneath the microchannel or underneath a parallel trough. The relative acoustic potential energy and the resultant velocity fields for particles with positive acoustic contrast coefficients are estimated in the 2-D limit. Confocal microscopy was used to observe the spatial distribution of the flowing microparticles in three dimensions. Through these studies, we show that a single driving frequency from a single piezoelectric actuator can induce the 2-D concentration of particles in a microchannel with a nearly square cross section, and we correlate these behaviors with theoretical predictions. We also show that it is possible to control the extent of focusing of the microparticles, and that it is possible to decouple the focusing of microparticles in the vertical direction from the lateral direction in rectangular channels with anisotropic cross sections. This study provides guidelines to design and operate microchip-based acoustofluidic devices for precise control over the spatial arrangement of microparticles for applications such as flow cytometry and cellular sorting.     

基于PPMgOLN晶体的高重频高效率中红外参量振荡器

设计了基于PPMgOLN晶体的高重频、高效率单谐振中红外参量振荡器。为了提高转化效率,自发研制了高功率,高重频,声光调Q的双端泵浦Nd:YVO4激光器作为泵浦光源,并对参量振荡腔进行了优化设计。在重复频率80 kHz,泵浦功率30 W时,获得了7 W的2.73μm中红外激光输出。光光转换效率为23.3%。采用温度调谐的方式,信号光输出光谱范围是1.67~1.75μm。对应的闲频光光谱范围是2.72~2.92μm。     

A microfluidic membrane device to mimic critical components of the vascular microenvironment

Vascular function, homeostasis, and pathological development are regulated by the endothelial cells that line blood vessels. Endothelial function is influenced by the integrated effects of multiple factors, including hemodynamic conditions, soluble and insoluble biochemical signals, and interactions with other cell types. Here, we present a membrane microfluidic device that recapitulates key components of the vascular microenvironment, including hemodynamic shear stress, circulating cytokines, extracellular matrix proteins, and multiple interacting cells. The utility of the device was demonstrated by measuring monocyte adhesion to and transmigration through a porcine aortic endothelial cell monolayer. Endothelial cells grown in the membrane microchannels and subjected to 20 dynes∕cm(2) shear stress remained viable, attached, and confluent for several days. Consistent with the data from macroscale systems, 25 ng∕ml tumor necrosis factor (TNF)-α significantly increased RAW264.7 monocyte adhesion. Preconditioning endothelial cells for 24 h under static or 20 dynes∕cm(2) shear stress conditions did not influence TNF-α-induced monocyte attachment. In contrast, simultaneous application of TNF-α and 20 dynes∕cm(2) shear stress caused increased monocyte adhesion compared with endothelial cells treated with TNF-α under static conditions. THP-1 monocytic cells migrated across an activated endothelium, with increased diapedesis in response to monocyte chemoattractant protein (MCP)-1 in the lower channel of the device. This microfluidic platform can be used to study complex cell-matrix and cell-cell interactions in environments that mimic those in native and tissue engineered blood vessels, and offers the potential for parallelization and increased throughput over conventional macroscale systems.     

污染环境中微生物培养模型的分析

恒化器是一个简单易于采用的用来培养微生物的实验装置。它被用做研究微生物的增长并有着对参数易于测量的优点。但是,关于恒化器模型的研究大都忽略了环境污染的情况。因此,本文考察在污染的环境中脉冲输入营养基Monod-Haldane恒化器模型．应用Floquet乘子理论和脉冲微分比较定理,得到微生物灭绝周期解是全局渐近稳定的充分条件,这意味着微生物培养失败。同时,得到在污染环境中微生物培养成功的条件,也就是系统持续生存的条件。最后讨论污染的环境对微生物培养的影响。     

Rapid production of protein-loaded biodegradable microparticles using surface acoustic waves

We present a straightforward and rapid surface acoustic wave (SAW) atomization-based technique for encapsulating proteins into 10 μm order particles composed of a biodegradable polymeric excipient, using bovine serum albumin (BSA) as an exemplar. Scans obtained from confocal microscopy provide qualitative proof of encapsulation and show the fluorescent conjugated protein to be distributed in a relatively uniform manner within the polymer shell. An ELISA assay of the collected particles demonstrates that the BSA survives the atomization, particle formation, and collection process with a yield of approximately 55%. The SAW atomization universally gave particles with a textured morphology, and increasing the frequency and polymer concentration generally gave smaller particles (to 3 μm average) with reduced porosity.     

Dielectrophoretic microfluidic device for the continuous sorting of Escherichia coli from blood cells

Microfluidic diagnostic devices promise faster disease identification by purifying and concentrating low-abundance analytes from a flowing sample. The diagnosis of sepsis, a whole body inflammatory response often caused by microbial infections of the blood, is a model system for pursuing the advantages of microfluidic devices over traditional diagnostic protocols. Traditional sepsis diagnoses require large blood samples and several days to culture and identify the low concentration microbial agent. During these long delays while culturing, the physician has little or no actionable information to treat this acute illness. We designed a microfluidic chip using dielectrophoresis to sort and concentrate the target microbe from a flowing blood sample. This design was optimized using the applicable electrokinetic and hydrodynamic theories. We quantify the sorting efficiency of this device using growth-based assays which show 30% of injected microbes are recovered viable, consistent with the electroporation of target cells by the dielectrophoretic cell sorters. Finally, the results illustrate the device is capable of a five-fold larger microbe concentration in the target analyte stream compared to the waste stream at a continuous sample flow rate of 35 μl∕h.