A microfluidic platform for formation of double-emulsion droplets

A new method for actively controlling the number of internal droplets of water-in-oil-in-water (W/O/W) double-emulsion droplets was demonstrated. A new microfluidic platform for double-emulsion applications has been developed, which integrates T-junction channels, moving-wall structures, and a flow-focusing structure. Inner water-in-oil (W/O) single-emulsion droplets were first formed at a major T-junction. Then the droplets were sub-divided into smaller uniform droplets by passing through a series of secondary T-junctions (branches). The moving-wall structures beside the secondary T-junctions were used to control the number of the sub-divided droplets by selectively blocking the branches. Finally, double-emulsion droplets were formed by using a flow-focusing structure downstream. Experimental data demonstrate that the inner and outer droplets have narrow size distributions with coefficient of variation (CV) of less than 3.5% and 5.7%, respectively. Double-emulsion droplets with 1, 2, 3, and up to 10 inner droplets have been successfully formed using this approach. The size of the inner droplets and outer droplets could be also fine-tuned with this device. The development of this new platform was promising for drug delivery applications involving double emulsions.     

A droplet-based microfluidic system capable of droplet formation and manipulation

Formation of emulsion droplets is crucial for a variety of industrial and scientific applications. This study presents a new droplet-based microfluidic system capable of generating tunable and uniform-sized droplets and subsequently deflecting these droplets at various inclination angles using a combination of flow-focusing and moving-wall structures. A pneumatic air chamber was used to activate the moving-wall structures, located nearby the outlet of the flow-focusing microchannels, such that the sheath flows can be locally accelerated. With this approach, the size of the droplets can be fine-tuned and sorted without adjusting the syringe pumps. Experimental data showed that droplets with diameters ranging from 31.4 to 146.2 μm with a variation of less than 5.39% can be generated. Besides, droplets can be sorted upwards or backwards with an inclination angle ranging from 0° to 53.5°. The development of this emulsion system may be promising for the formation and collection of emulsion products for applications in the pharmaceutical, cosmetics and food industries.     

A compact microfluidic gradient generator using passive pumping

Creating and maintaining a precise molecular gradient which is stable in space and time are essential to studies of chemotaxis. This paper describes a simple, compact, and user-friendly microfluidic device using a passive pumping method to drive liquid flow to generate a stable concentration gradient. A fluidic circuit is designed to offset the effects of the pressure imbalance between the two inlets. After loading approximately the same amount of culture media containing different concentrations of a certain chemotactic agent into the two inlet reservoirs, a linear concentration gradient will be automatically and quickly established at the downstream. Our device takes advantage of passive pumping and is compact enough to fit into a Petri dish, which is an attractive feature to biologists. Furthermore, this microfluidic gradient generator offers a platform for a facile way of long-term imaging and analysis using high-resolution microscopy.     

微沟道内两相流速比对液滴形成的影响

研究了流聚焦微沟道内不同两相流速比对单分散性液滴形成的影响.不相混容的两相系统能够快速、周期性地形成液滴,液滴的形态和生成率随着油相速度的变化而改变.研究采用模拟软件COMSOL Mutiphysics,模拟了不同流速下液滴的形成过程,并通过实验进行了验证,从流体动力学的角度解释了其物理机制.所得结论为在流聚焦微沟道内获得分散性良好且大小可控的液滴提供了理论和数据支持.     

A microfluidic chip for heterogeneous immunoassay using electrokinetical control

This article presents the development of a novel, automated, electrokinetically controlled heterogeneous immunoassay on a poly(dimethylsiloxane) (PDMS) microfluidic chip. A numerical method has been developed to simulate the electrokinetically driven, time-dependent delivery processes of reagents and washing solutions within the complex microchannel network. Based on the parameters determined from the numerical simulations, fully automated on-chip experiments to detect Helicobacter pylori were accomplished by sequentially changing the applied electric fields. Shortened assay time and much less reagent consumptions are achieved by using this microchannel chip while the detection limit is comparable to the conventional assay. There is a good agreement between the experimental result and numerical prediction, demonstrating the effectiveness of using CFD to assist the experimental studies of microfluidic immunoassay.     

Physical stimulation of mammalian cells using micro-bead impact within a microfluidic environment to enhance growth rate

Various cell stimulation experiments have been traditionally conducted regarding observation of cells to specific stimulative factors, and in light of this area of study, we report a new method by utilizing micro-beads. HeLa cells and MC3T3 cells are cultured in straight PDMS (polydimethylsiloxane) microfluidic devices and are stimulated by sterilized polystyrene micro-beads. Cell culture medium either with or without micro-beads are introduced in microfluidic cell culturing chambers at a specific time interval, and stimulated cells are observed using an inverted microscope. The results show that cells exposed under micro-bead stimulation perform at a higher growth rate than those under normal conditions. This paper demonstrates that micro-beads can be used as a physical stimulation factor and affect cell growth behaviors.     

A novel microfluidic switch for pH control using Chitosan based hydrogels

We report a novel pH-sensitive hydrogel based micro-valve for metered flow that has applications in a laboratory made “Intelligent valving system”. The hydrogel solution was prepared through Chitosan and poly vinyl alcohol in acetic acid and crystallized using gluteraldehyde as the crosslinking agent in the form of thin wafers and it was found to be very sensitive to pH changes. The pore structure of hydrogel was investigated through Field Emission Scanning Electron Microscopy and thin wafers of the gel were physically placed inside PDMS microchannels. Flow metering in these channels was observed by controlled expansion of the hydrogel plug till complete valving was realized. This valving device was further precisely characterized with micro Particle Image Velocimetry using a solution containing fluorescent polymeric micro beads. The principle advantage of this hydrogel device is the smaller range of pH (varying between pH 3 and 7) over which the valving response is observed.     

A novel three-dimensional microfluidic platform for on chip multicellular tumor spheroid formation and culture

The formation of three-dimensional (3D) multicellular cell spheroids such as microspheres and embryoid bodies has recently gained much attention as a useful cell culture technique, but few studies have investigated the suitability of glass for spheroids formation and culture. In this work, we present a novel three-dimensional microfluidic device made of poly(dimethylsiloxane) (PDMS) and glass for the easy and rapid synthesis and culture of tumor spheroid. The cell culture unit is composed of an array of microwells on the bottom of a glass plate, bigger microwells and elastomeric microchannels on the top of a PDMS plate. Cell suspension can be easily introduced into the cell culture unit and exchange with the external liquid environment by the microfluidic channels. A single tumor spheroid can be formed and cultured in each glass cell culture chamber, the surface of which was modified with poly(vinyl alcohol) to render it to be resistant to cell adhesion. As the cell culture medium could be replaced, spheroids of the human breast cancer (MCF-7) cells were cultured on the chip for 3 days, reaching the diameters of about 150 μm. Furthermore, the MCF-7 cells were successfully cultured on the chip in 2D and 3D culture modes. Results have shown that glass is well suitable for multicellular tumor spheroids culture. The established platform provides a convenient and rapid method for tumor spheroid culture, which is also adaptable for anticancer drug screening and fundamental biomedical research in cell biology.     

In situ kinetics study of the formation of organic nanoparticles by fluorescence lifetime imaging microscopy (FLIM) along a microfluidic device

In this study, a three-dimensional hydrodynamic focusing microfluidic method is presented that allows full control of the nano-precipitation process of adamantyl mesityl BODIPY (4,4-difluoro-3,5-di-(adamantyl)-8-mesityl-4-bora-3a,4a-diaza-s-indacene) (Adambodipy). The precipitation is achieved by combining a central Adambodipy organic flow with a mixture of water and a cationic surfactant, creating a non-solvent precipitation method. The flow and mixing were simulated using COMSOL Multiphysics^® 3.4. A good agreement between theory and experiment was obtained for the flow velocity, concentration fields and the subsequent precipitation kinetics. Fluorescence lifetime imaging was used to visualize the precipitation domains following the changes in fluorescence lifetime. The lifetime decreases from 6.1 ns for the molecules down to 0.9 ns for nanoparticles. A principal components analysis of the successive fluorescence decay curves showed that the process could be adequately modeled using three components, which can be attributed to monomers (single molecule), clusters (nuclei) and nanoparticles.     

Globally stabilizing control of fed-batch processes with Haldane kinetics using growth rate estimation feedback

The regulation of the biomass specific growth rate is an important goal in many biotechnological applications. To achieve this goal in fed-batch processes, several control strategies have been developed employing a closed loop version of the exponential feeding law, an estimation of the controlled variable and some error feedback term. In the case of non-monotonic kinetics, the specified growth rate can be achieved at two different substrate concentration values. Because of the inherent unstable properties of the system in the decreasing portion of the kinetics function, stabilization becomes a crucial problem in this high-substrate operating region. In this context, the dynamic behavior of fed-batch processes with Haldane kinetics is further investigated. In particular, some conditions for global stability and performance improvement are derived. Then, a stabilizing control law based on a partial state feedback with gain dependent on the output error feedback and gain saturation is proposed. Although particular emphasis is put on the critical case of high-substrate operation, low-substrate regulation is also treated.     

A method of water pretreatment to improve the thermal bonding rate of PMMA microfluidic chip

A new method of water pretreatment for thermal bonding polymethylmethacrylate microfluidic chip was proposed in this paper. The bonding rate (effective bonding area) of microfluidic chip under different pretreatment time was studied and the mechanism of this method was discussed. The main thermal bonding parameters were as follows: bonding pressure 1.4 ~ 1.9 Mpa, temperature 91 ~ 93°C, time 360 s. The experimental result shows that this method can increase the effective bonding area, improve the bonding quality of the microfluidic chip compared to the conventional thermal bonding method. The optimal water pretreatment time is 1 h with the bonding rate increased by 34% compared with the conventional thermal bonding method. The pollution to the micro-channels is avoided and the performance of the microfluidic system will be reserved with this water pretreatment method. This method is available for the biochemical analysis of the chip, and holds the benefits of easy-operation, high-efficiency and low-cost properties.     

A simple photolithography method for microfluidic device fabrication using sunlight as UV source

A straightforward method for microfluidic devices fabrication using sunlight as the ultraviolet (UV) source is established in this work. This method is based on photolithography, but obviates the need for specialized UV exposure facility. Substrates coated with photoresist were placed directly under sun in a perpendicular direction to the sunlight for exposure. Exposure conditions were optimized for patterning features with different kinds of photoresist, photoresist of different thicknesses and dimensions. Exposure time can be adjusted to obtain designed features on a mask with good lateral structure according to the energy measured by UV meter (with a constant intensity of UV in sunlight). Masters produced under optimum exposure conditions were used for the fabrication of several microfluidic devices with different materials, structures, or functions. Resultant devices were shown eminently suitable for microfluidic applications such as electrophoretic separation, multiple gradient generator, and pneumatic valve-based cell culture. This photolithographic method is simple, low cost, easy to operate, and environmental friendly. Especially, the masters can be obtained in parallel simultaneously, which is suitable for chip fabrication for mass production. It is also more attractive for the laboratories, in which the support for photolithographic facility is not available.     

A liquid rate gyroscope using electro-conjugate fluid

We propose a novel liquid rate gyroscope using an electro-conjugate fluid (ECF). The electro-conjugate fluid is a dielectric fluid that works as a smart fluid, generating a powerful jet flow (ECF jet) when subjected to a high DC voltage. In this study, we introduce this functional fluid into gyroscopes. Although the sensing principle for angular rate is based on that of a conventional gas rate sensor, the proposed gyroscope has a much higher sensitivity because the density of the liquid is generally higher than that of a gas. In addition, the gyroscope is small in size because the ECF jet is generated only with a pair of tiny electrodes. In other words, the pumping part of the proposed gyroscope does not need mechanical moving parts, resulting in an ECF gyroscope more suitable for micro-applications than a gas rate sensor, which requires a pumping mechanism inside. We fabricated a prototype of the liquid rate gyroscope (40 mm × 60 mm × t7 mm) and confirmed its characteristics by experiments. The experimental results confirm the effectiveness of the proposed liquid rate gyroscope. The prototype has a scale factor of ?29 mV/(°/s) with an applied voltage of 4.5 kV, which is 2.2 times more sensitive than the conventional gas rate gyroscope.     

A digital dilution chip using inter-well valves controlled by a ternary microfluidic multiplexer

We present a digital dilution chip of 2 × 6 well array that is capable of changing the dilution ratio. We mixed the diluted samples with other samples by using inter-well valves. The previous continuous and digital dilution chips diluted samples that had a fixed dilution ratio depended on the structure of the branched microchannel or the volume of the well. The present chip can perform the programmable mixing process that selectively fills, merges, and splits the identical wells by controlling the inter-well valves. Thus, the present digital dilution chip can easily change the dilution ratio without any structural change. In this study, the present chip changed the dilution ratio, such as the linear or exponential ratio, within a 4.2% dilution difference. They were then mixed with other samples within a 3.4% mixing difference. In order to reduce the number of interconnection ports that transferred the pressure to inter-well valves, we also applied a latched ternary multiplexer and reduced nine interconnection ports of inter-well valves to five. By diluting the sample with different dilution ratios based on the well array, the present chip can be easier for users and more suitable to high-throughput screen systems.     

A nonlinear least-squares method for determining cerebrospinal fluid formation and absorption kinetics in pseudotumor cerebri

Data from 14 patients with benign intracranial hypertension (pseudotumor cerebri) have been analyzed using a nonlinear least-squares regression model which was developed and programmed from in-hospital microcomputer use. The method of analysis permits rapid estimation of cerebrospinal fluid (CSF) formation and absorption rates as functions of pressure in individual patients using data from constant-rate infusion manometrics. The analysis predicts that prednisone therapy in pseudotumor cerebri reduces resting CSF pressure by increasing CSF absorption at all intracranial pressures studied, and decreasing CSF formation at high pressures. This result is in accordance with evidence suggesting that impaired CSF absorption plays a major role in the pathogenesis of increased intracranial pressure in pseudotumor cerebri.     

A program for the optimization of cyclosporine therapy using population kinetics modeling

Cyclosporine is one of the most widely used immunosuppressive agents in organ transplantation. Due to large inter- and intra-individual variations, its behavior in the specific patient is still difficult to predict. Dosage optimization is thus mainly performed on a trial-and-error basis. In this paper, we present a new program based on the population kinetics approach, which was designed to help physicians in the difficult task of adjusting patient specific cyclosporine dosing regimens. Dose optimization is carried out by model simulation, using a two-compartment mathematical model of cyclosporine kinetics to predict the drug behavior in the patient. Two of the model parameters are assumed from the literature, the other two are estimated from the patient data through a Bayesian estimation procedure. Previous information needed by the Bayesian algorithm is derived by a population analysis, performed beforehand and based on a nonlinear mixed effect model. A user-friendly graphical interface written in Delphi under Windows makes the program easily accessible to physicians. A preliminary retrospective validation of the program, performed on data from 18 renal transplanted patients, yielded very satisfactory results.     

A microfluidic gel valve device using reversible sol–gel transition of methyl cellulose for biomedical application

We have fabricated a microfluidic gel valve device that used reversible sol–gel transition of methyl cellulose (MC). A microheater and a microtemperature sensor were implemented in each microchannel in the gel valve device. Before evaluating the performance of the gel valve device, various properties of the MC solution were investigated using viscometer, spectrophotometer, and NMR. Gelation temperature was increased as the MC concentration was increased. Clear gel, an intermediate state between clear sol and turbid gel, was found at the temperature range from 30–40°C to 50–60°C. Temperature at each microchannel of the device was measured and the effect of the temperature difference on the valve operation was elucidated. In order to have normal operation of the gel valve, it was important to keep the temperature of the heated microchannel around 60°C while keeping the temperature of the flowing microchannel below 35°C. The temperature difference between two microchannels was about 23 K when fan forced cooling (FFC) method was used. For normal performance of the gel valve device, a temporary pause of fluid flow for at least 5 s was required to complete the local gelation in the microchannel. Stable gel valve performance was obtained at the flow rates larger than 5 μl/min. The gel valve device showed no leakage up to 2.07×10⁴ Pa.     

A highly efficient and reliable heart rate monitoring system using smartphone cameras

Multimedia Tools and Applications - In the field of healthcare, a fundamental research topic includes the development of a vital signs monitoring system that allows people to monitor their health...