A microfluidic-based dynamic microarray system with single-layer pneumatic valves for immobilization and selective retrieval of single microbeads

A simple yet effective dynamic bead-based microarray is necessary for multiplexed high-throughput screening applications in the fields of biology and chemistry. This paper introduces a microfluidic-based dynamic microbead array system using pneumatically driven elastomeric valves integrated with a microchannel in a single polydimethylsiloxane (PDMS) layer that performs the following functions: single-microbead arraying with loading and trapping efficiencies of 100 %, sequential microbead release for selective retrieval of microbeads of interest, and rapid microarray resettability (<1 s). The key feature is the utilization of an elastomeric membrane as a valve for trapping and releasing single microbeads; this membrane is deformable depending on the applied pneumatic pressure, thereby simply providing a dual trap-and-release function. We propose an effective single-microbead-trapping mechanism based on a dynamic flow-change network and a mathematical model as the design criterion of a trapping site. A sequential microbead release technique via a multistep “release-retrap-and-repeat” method was developed for the selective retrieval of trapped microbeads with a simple configuration consisting of a single PDMS layer and a simple macro-to-micro connection. The proposed dynamic microbead array could be a powerful tool for high-throughput multiplex bead-based drug screening or disease diagnosis.     

Detection of microdroplet size and speed using capacitive sensors

Detection of the presence, size and speed of microdroplets in microfluidic devices is presented using commercially available capacitive sensors which make the droplet based microfluidic systems scalable and inexpensive. Cross-contamination between the droplets is eliminated by introducing a passivation layer between the sensing electrodes and droplets. A simple T-junction generator is used to generate droplets in microchannels. Coplanar electrodes are used to form a capacitance through the microfluidic channel. The change in capacitance due to the presence of a droplet in the sensing area is detected and used to determine the size and speed of the droplet. The design of a single pair of electrodes is used to detect the presence of a droplet and the interdigital finger design is used to detect the size and speed of the droplet. An analytical model is developed to predict the detection signal and guide the experimental optimization of the sensor geometry. The measured droplet information is displayed through a Labview interface in real-time. The use of capacitance sensors to monitor droplet sorting at a T-junction is also presented. The discussions in this paper can be generalized to any droplet detection application and can serve as a guideline in sensor selection.     

Development of microfluidic alginate microbead generator tunable by pulsed airflow injection for the microencapsulation of cells

Recently, microbead generation and microencapsulation of cells using microfluidic technology have been actively pursued for various applications. However, most of the proposed systems are not only technically demanding, but might also be harmful to the encapsulated cells. To tackle these issues, this study reports a microfluidic alginate microbead generator consisting of a polydimethylsiloxane (PDMS) microfluidic chip and an integrated quartz microcapillary tube. The working principle is based on the use of a pulsed airflow to segment a continuous alginate suspension flow to form suspension fragments in a microchannel and then alginate microbeads when they were delivered out the microfluidic system to a sterile calcium chloride solution through a microcapillary tube. In this study, the alginate suspension fragments with varied sizes in the microchannel can be generated either by modulating the alginate suspension flow rate or the pulsation frequency of airflow injection. By fine tuning the size of them, the alginate microbeads can be generated in a size-controllable manner. Results showed that alginate microbeads with the size ranging from 150 to 370 μm in diameter can be generated at the suspension flow rate and airflow injection frequency ranges of 2–4 μl/min and 0.6–35 Hz, respectively. Besides, the alginate microbeads generated by the system were tested with excellent size uniformity (CV: 3.1–5.1%). Moreover, its application for the microencapsulation of chondrocytes in alginate microbeads was also demonstrated with high cell viability (96 ± 2%). As a whole, the proposed device has paved an alternative route to perform alginate microbead generation or microencapsulation of cells in a simple, continuous, controllable, uniform, cell friendly, and less contaminated manner.     

Au纳米粒子修饰纳米片状结构衬底的SERS研究

通过电化学共沉积和化学脱合金处理在金属W片上制备了纳米片状基底,将Au纳米粒子通过等离子溅射到纳米片状基底上得到表面增强拉曼散射(SERS)衬底。采用扫描电子显微镜(FE-SEM)、能谱仪(EDX)对复合纳米衬底进行表征,罗丹明6G作为探测分子对SERS衬底的拉曼表面增强效果进行检测。通过实验发现:三维空间结构的纳米片状结构具有拉曼表面增强效应,溅射Au纳米颗粒得到的Au纳米片衬底信号增强效果显著。     

Driving and sorting of the fluorescent droplets on digital microfluidic platform

In this paper, we present a digital microfluidic droplet sorting platform to achieve automated droplet sorting based on fluorescent detection. We design and fabricate a kind of digital microfluidic chip for manipulating nano-liter-sized liquid droplets, and the chip is integrated with a fluorescence-initiated feedback system for real-time sorting control. The driving and sorting characteristics of fluorescent droplets encapsulating fluorescent-labeled particles are studied on this platform. The droplets dispensed from on-chip reservoir electrode are transported to a fluorescence detection site and sorted according to their fluorescence signals. The fluorescent droplets and non-fluorescent droplets are successfully separated and the number of fluorescent particles inside each droplet is quantified by its fluorescent intensity. We realize droplet sorting at 20 Hz and obtain a linear relationship between the fluorescent particle concentrations and the fluorescence signals. This work is easily adapted for sorting out fluorescent-labeled microparticles, cells and bacteria and thus has the potential of quantifying catalytic or regulatory bio-activities.     

喷射式3D打印过程微滴沉积位置建模与预测

微滴沉积位置的准确性是影响喷射成型器件形貌精度的主要因素之一,为了对微滴沉积位置进行控制,需要建立微滴沉积位置的动态预测模型。建立了微滴在下落、沉积过程中的运动与传热模型,并通过数值仿真和实验研究了单个微滴以及两个相邻微滴在沉积动态过程中的形心位置变化,结果表明,相比于数值仿真模型,所建立的理论预测模型对单个微沉积位置预测误差为0.45%,相邻两个微滴间的沉积距离预测误差为0.5%,并通过实验进行了对比验证,表明所建理论模型具有足够高的位置预测精度,可用于微滴喷射3D打印过程中微滴沉积位置及打印轨迹在线控制的参考。     

Hydrodynamic trap-and-release of single particles using dual-function elastomeric valves: design, fabrication, and characterization

This paper introduces a simple method for trapping and releasing single particles, such as microbeads and living cells, using dual-function elastomeric valves. Our key technique is the utilization of the elastomeric valve as a dual-function removable trap instead of a fixed trap and a separate component for releasing trapped particles, thereby enabling a simple yet effective trap-and-release of particles. We designed, fabricated, and characterized a microfluidic-based device for trapping and releasing single beads by controlling elastomeric valves driven by pneumatic pressure and a fluid flow action. The fluid flow is controlled to ensure that beads flowing in a main stream enter into a branch channel. A bead is trapped by deflected elastomeric valves positioned at the entrance of a branch channel. The trapped bead is easily released by removing the applied pressure. The trapping and releasing of single beads of 21?μm in diameter were successfully performed under an optimized pressure and flow rate ratio. Moreover, we confirmed that continuous trapping and releasing of single beads by repeatedly switching elastomeric valves enables the collection of a controllable number of beads. Our simple method can be integrated into microfluidic systems that require single or multiple particle arrays for quantitative and high-throughput assays in applications within the fields of biology and chemistry.     

Creation of single-particle environment by positive dielectrophoresis and liquid dielectrophoresis

Two electrical mechanisms for manipulating particles and fluids, dielectrophoresis (DEP) and liquid dielectrophoresis (LDEP), are integrated in a microfluidic chip for creating the single-particle environment. The fluid is activated by LDEP with a 100-kHz/240-V_pp signal. When the single polystyrene bead approaches the trapping area, positive DEP force is utilized to capture and immobilize the bead. After trapping the bead, the process of liquid cutting and droplet creation is employed to create a droplet containing a single bead by LDEP with a 100-kHz/320-V_pp signal.     

Frequency-dependent conductance change of dielectrophoretic-trapped DNA-labeled microbeads and its application in DNA size determinations

DNA amplification is essential in several types of molecular biology approaches. A more rapid and easy analysis of amplicons is still required although many analysis methods have been developed. We have recently devised a new DNA detection method, where DNA amplicons are attached to dielectric microbead surfaces, so that their dielectrophoresis (DEP) force on the microbead reverses polarity, from negative to positive. The DNA-labeled microbeads are trapped on a microelectrode by positive DEP, enabling their rapid detection via DEP impedance measurement. In this paper, we report frequency-dependent conductance of DNA-labeled microbeads. To measure the impedance, sweep-frequency voltage was superimposed on fixed-frequency voltage, with the aim of inducing frequency-dependent conformational change of microbead-attached DNA, ultimately resulting in a change in the conductance of DNA-labeled microbeads. Microbeads labeled with DNA of various sizes (142-, 204-, 391-, and 796-bp) were examined. The normalized conductance sharply decreased at a specific frequency; the frequency was higher with larger DNA size, suggesting a potential application of this method in distinguishing DNA targets according to their size. By combining this method with previously devised DNA detection techniques, both the size and amount of target DNA can be determined within 20 min. This approach is easier and more rapid than conventional methods, such as a gel electrophoresis.     

Optical measurement of flow field and concentration field inside a moving nanoliter droplet

Droplets-based method has been employed to enhance mixing in microfluidic systems. This paper presents experimental studies of the recirculating flow field inside a moving droplet and the characterization of the mixing of two aqueous droplets. In the first part, the velocity field inside the moving water droplet was measured using the micro-particle image velocimetry (micro-PIV) technique. The PIV measurements showed that recirculation flow exists inside the droplet. However, the findings suggested that the outer layer of droplets move at a faster velocity than the central part. The result is different from what is reported by other researchers. In the second part, two water droplets, a de-ionized (DI) water droplet and another DI water droplet with fluorescent dye, were brought together by the carrier fluid to form a bigger droplet. The mixing between the two aqueous droplets was characterized by the fluorescent dye concentration distribution.     

A particle swarm optimization method for fault localization and residue removal in digital microfluidic biochips

Second generation microfluidic biochip is known as digital microfluidic biochip (DMFB). DMFB performs different clinical pathological experimentation, DNA sequencing, air contamination detection and many other bio-chemical experiments based on appropriate bio-assay protocols. DMFB comprises two dimensional array of electrodes fabricated over two parallel glass plates, which is capable of performing miniaturized (nano/ pico liter volume) droplet dispensing, transportation and mixing through electro-wetting of dielectrics (EWOD). Droplet operations through EWOD are mostly managed by droplet scheduling algorithms which are NP hard in nature. At present reliability is a major concern for commercialization of operational DMFB. Reliable output of DMFB is mostly affected by different faults within electrodes. This further suffers from cross-contamination issues among different droplets due to repeated use of DMFB boards for different assay operations. Present work proposes a novel test droplet routing method based on adaptive weighted particle swarm optimization (PSO) model. This test droplet circulation method aims to identify defective electrodes and simultaneously performs residue removal. Experimental findings of the proposed model on some standard test benches and real life bio-assay samples reveal operational supremacy in terms of overall computational time and operational accuracy over some existing best known models.     

Reliable addition of reagents into microfluidic droplets

This article reports a design that reliably adds reagents into droplets by exploiting the physics of fluid flow at a T-junction in the microchannel. An expanded section right after the T-junction enhances merging of a stream with a droplet, eliminates the drawbacks such as extra droplet formation and long mixing time. The expanded section reduces the pressure buildup at the T-junction and minimizes the tendency to form extra droplets; plays the role in creating low Laplace pressure jump across the interface of the droplet forming from the T-junction which reduces the probability of forming extra droplet in the merging process; provides space for droplet coalescence if there is an extra droplet due to droplet break-up before merging. In this design, after merging, the reactants are in axial arrangement inside the droplets which lead to faster mixing. Reliable addition of reagent to the droplets happens for the combination of flow rates in a broad range from 25 to 250 μl/h, for both DI water (Q _DI) and fluorescent (Q _fluo) streams.     

Kinematics and deformation of ferrofluid droplets under magnetic actuation

This paper reports the experimental results on kinematics and deformation of ferrofluid droplets driven by planar coils. Ferrofluid droplets act as liquid magnets, which can be controlled and manipulated by an external magnetic field. In our experiments, the magnetic field was generated by two pairs of planar coils, which were fabricated on a double-sided printed circuit board. The first pair of coils constrains the ferrofluid droplet to a one-dimensional motion. The second pair generates the magnetic gradient needed for the droplet motion. The direction of the motion can be controlled by changing the sign of the gradient or of the driving current. Kinematic characteristics of the droplet such as the velocity–position diagram and the aspect ratio of the droplet are investigated. The analysis and discussion are based on the different parameters such as the droplet size, the viscosity of the surrounding medium, and the driving current. This simple actuation concept would allow the implementation of lab-on-a-chip platforms based on ferrofluid droplets.     

A MEMS Thermal Biosensor for Metabolic Monitoring Applications

This paper presents a microelectromechanical systems (MEMS) differential thermal biosensor integrated with microfluidics for metabolite measurements in either flow-injection or flow-through mode. The MEMS device consists of two identical freestanding polymer diaphragms, resistive heaters, and a thermopile between the diaphragms. Integrated with polymer-based microfluidic measurement chambers, the device allows sensitive measurement of small volumes of liquid samples. Enzymes specific to a metabolic analyte system are immobilized on microbeads packed in the chambers. When a sample solution containing the analyte is introduced to the device, the heat released from the enzymatic reactions of the analyte is detected by the thermopile. The device has been tested with glucose solutions at physiologically relevant concentrations. In flow-injection mode, the device demonstrates a sensitivity of approximately 2.1 muV/mM and a resolution of about 0.025 mM. In flow-through mode with a perfusion flow rate of 0.5 mL/h, the sensitivity and resolution of the device are determined to be approximately 0.24 muV/mM and 0.4 mM, respectively. These results illustrate that the device, when integrated with subcutaneous sampling methods, can potentially allow for continuous monitoring of glucose and other metabolites.     

Droplet bistability and its application to droplet control

Novel methods for controlling droplets precisely in the microchannels are presented, which employ microfluidic bifurcation channels with outlet restrictions based on droplet bistability, utilizing the Laplace pressure caused by interfacial tension that arises when a droplet encounters a narrow restriction. The bistable geometry possesses two symmetric branches and restrictions that operate as capillary valves allowing a droplet to be trapped in front of a restriction and released through it when the next droplet arrives at the other restriction. This trap-and-release occurs repeatedly and regularly by the succeeding droplets. Furthermore, a critical flow rate is found to exist, which is necessary for achieving droplet bistability. This occurs only when the apparent Laplace pressure surpasses the pressure drop across the droplet. By adopting a simplified hydrodynamic resistance model, the droplet bistable mechanism is clearly explained, and droplet bistability is shown to enable the simple and precise control of droplets at a bifurcation channel. Thus, precise droplet traffic control is achieved at a bifurcation channel connected with a single inlet channel and two outlet channels using an appropriate channel design that induces droplet bistability. In particular, droplets are distributed at a junction in a manner of perfect alternation or perfect switching between the two outlet channels. This article proposes that bistable components can be used as elaborately embedded droplet traffic signals for red light (trap), green light (release), and turn light (switching) in complex microfluidic devices, where droplets provide both the chemical or biological materials and the processing signal.     

聚苯胺-多壁碳纳米管薄膜SAW NO2传感器

将聚苯胺-多壁碳纳米管聚合物沉积于声表面波(SAW)气体传感器的敏感区,形成敏感薄膜,实现了室温下对不同体积分数NO2的检测.研究表明:这种传感器具有很高的敏感性,良好的重复性和低体积分数检测极限.同时,单一聚苯胺薄膜也做了对比测量,实验结果表明:聚苯胺-多壁碳纳米管聚合物敏感膜比单一聚苯胺薄膜具有更高的敏感性和更短的...     

Generation of droplets with different concentrations using gradient-microfluidic droplet generator

This study successfully uses the micro-mixers and flow-focusing devices, which are integrated into a gradient-microfluidic droplet generator, to generate the different sizes of the droplets with different concentrations simultaneously and applies these microcapsules for drug release. The sizes of these four types of droplet with different concentrations are uniformity with a coefficient of variation less than 5% and can be precisely controlled by adjusting the water phase flow rate and oil phase flow rate. Moreover, Ca-alginate microcapsules with different concentrations of the bovine serum albumin are used for uniform size drug release, and the Ca-alginate microcapsule size is from 60 to 105 μm in diameter. This developed microfluidic chip has the advantages of actively controlling the droplet diameter, simultaneously generating uniform-sized droplets with different concentrations, and having a simple process and a high throughput. This preparation approach for Ca-alginate microcapsules of four different concentrations will provide many potential applications for drug delivery and pharmaceutical area.     

Droplet formation behavior in a microfluidic device fabricated by hydrogel molding

We describe the behavior of droplet formation within 3D cross-junctions and 2D T-junctions with various cross-sectional geometries that were manually fabricated using the hydrogel-molding method. The method utilizes wire-shaped hydrogels as molds to construct 3D and 2D microchannel structures. We investigated the flow patterns and droplet formation within the microchannels of these microfluidic devices. Despite being fabricated manually, the microchannels with 3D cross-junctions and 2D T-junctions were reproducible and formed highly monodispersed droplets. Additionally, the sizes of the droplets formed within the microchannels could be predicted using an experimental formula. This technique of droplet formation involves the use of a device fabricated by hydrogel molding. This method is expected to facilitate studies on droplet microfluidics and promote the use of droplet-based lab-on-a-chip technologies for various applications.     

Ferrofluid-molding method for polymeric microlens arrays fabrication

This study proposes a method named as ferrofluid-molding method for polymer microlens array fabrication. In this method, the master of the mother mold for microlens molding is an array of ferrofluid droplets. We generated droplet arrays by inducing the droplet’s magnetic hydrodynamic instability under different magnetic fields, and used the field-dependent droplet dimensions to fabricate numerous mold cavities. By this we could fabricate arrays of microlens with different bottom area, height, radius of curvature, and focal length. From our analysis, all the fabricated microlens arrays possessed good uniformity, and the largest numerical aperture of our microlens array was found as 0.54. In addition, we also designed a light uniformity experiment to demonstrate a potential application of our microlens arrays.     

Microfabricated micropallets for enhancement of biomolecular techniques

We introduce microcarriers referred to as “micropallets” ranging in size from 25 μm to several 100 microns, fabricated using photoresist or other polymer materials. These small carrier structures may be used in static detection systems or for transporting attached biological or chemical samples through a microfluidic system. Micropallets may be encoded through the use of barcodes or other markings and engineered to optimally suit the cargo they carry. We demonstrate the use of micropallets in cell and antibody assays. Furthermore we demonstrate the ability to decode and manipulate micropallets in a flow-through system.