PDMS-based Microfluidic Device Research Articles

Three-dimensional magnetic focusing of particles and cells in ferrofluid flow through a straight microchannel

Focusing particles and cells into a tight stream is often required in order for continuous flow detection, counting and sorting. So far a variety of particle focusing methods have been developed in microfluidic devices, among which magnetic focusing is still relatively new. We develop in this work an approach to embedding symmetrically two repulsive permanent magnets about a straight rectangular microchannel in a PDMS-based microfluidic device. The closest distance between the magnets is limited only by the sizes of the embedded and holder magnets involved in the fabrication process. The developed device is used to implement and investigate the three-dimensional magnetic focusing of polystyrene particles in ferrofluid microflow with both the top- and side-view visualizations. The effects of flow speed and particle size on the particle focusing effectiveness are studied. The developed device is also applied to magnetically focus yeast cells in ferrofluid, which proves to be biocompatible as verified by a cell viability test. In addition, an analytical model is developed and found to be able to predict the experimentally observed particle and cell focusing behaviors with reasonable agreement.

A “place n play” modular pump for portable microfluidic applications

This paper presents an easy-to-use, power-free, and modular pump for portable microfluidic applications. The pump module is a degassed particle desorption polydimethylsiloxane (PDMS) slab with an integrated mesh-shaped chamber, which can be attached on the outlet port of microfluidic device to absorb the air in the microfluidic system and then to create a negative pressure for driving fluid. Different from the existing monolithic degassed PDMS pumps that are generally restricted to limited pumping capacity and are only compatible with PDMS-based microfluidic devices, this pump can offer various possible configures of pumping power by varying the geometries of the pump or by combining different pump modules and can also be employed in any material microfluidic devices. The key advantage of this pump is that its operation only requires the user to place the degassed PDMS slab on the outlet ports of microfluidic devices. To help design pumps with a suitable pumping performance, the effect of pump module geometry on its pumping capacity is also investigated. The results indicate that the performance of the degassed PDMS pump is strongly dependent on the surface area of the pump chamber, the exposure area and the volume of the PDMS pump slab. In addition, the initial volume of air in the closed microfluidic system and the cross-linking degree of PDMS also affect the performance of the degassed PDMS pump. Finally, we demonstrated the utility of this modular pumping method by applying it to a glass-based microfluidic device and a PDMS-based protein crystallization microfluidic device.

TiO 2 nanofibers fixed in a microfluidic device for rapid determination of chemical oxygen demand via photoelectrocatalysis

Large arrays of one-dimensional uniform-sized TiO 2 nanofibers (TNFs) were prepared through a template-free method, and used as a working electrode in a transparent microfluidic device made from poly (dimethyl siloxane) (PDMS) to perform efficient photoelectrocatalysis for rapid and undefiled determination of chemical oxygen demand (COD). Photoelectrochemical measurements were used to evaluate the response of TNFs to the intensity of exciting light and the applied potential bias. The photoelectrocatalysis of TNFs in PDMS-based microfluidic device exhibited excellent performance for determination of COD. The practical limit of determination of 0.95 mg/L COD with a working range of 0–250 mg/L was achieved. The relative standard deviation (RSD) was 1.85% for 10 repeated measurements of 0.3 mmol/L glucose with COD value of 57.6 mg/L.

A microfluidic-based hydrodynamic trap: design and implementation

We report an integrated microfluidic device for fine-scale manipulation and confinement of micro- and nanoscale particles in free-solution. Using this device, single particles are trapped in a stagnation point flow at the junction of two intersecting microchannels. The hydrodynamic trap is based on active flow control at a fluid stagnation point using an integrated on-chip valve in a monolithic PDMS-based microfluidic device. In this work, we characterize device design parameters enabling precise control of stagnation point position for efficient trap performance. The microfluidic-based hydrodynamic trap facilitates particle trapping using the sole action of fluid flow and provides a viable alternative to existing confinement and manipulation techniques based on electric, optical, magnetic or acoustic force fields. Overall, the hydrodynamic trap enables non-contact confinement of fluorescent and non-fluorescent particles for extended times and provides a new platform for fundamental studies in biology, biotechnology and materials science.

High-speed droplet generation on demand driven by pulse laser-induced cavitation

We report on a pulse laser-driven droplet generation (PLDG) mechanism that enables on-demand droplet generation at rates up to 10,000 droplets per second in a single-layer PDMS-based microfluidic device. Injected droplet volumes can be continuously tuned between 1 pL and 150 pL with less than 1% volume variation.

Microfluidic Preparation of Dual Stimuli-Responsive Microparticles and Light-Directed Clustering

We present a simple fabrication of photo- and thermoresponsive microparticles with a narrow size distribution in the PDMS-based microfluidic device. The monodisperse water-in-oil (W/O) droplets of poly(N-isopropylacrylamide-co-spironaphthoxazine methacryloyl) (PNIPA-SPO) were formed at the T-junction channel of the device by adjusting the flow conditions of two immiscible solutions. Subsequently, the droplets were polymerized downstream of the channel under 365 nm UV irradiation in the presence of 2,2'-diethoxyacetophenone (DEAP, photoinitiator) and N,N'-methylenebisacrylamide (MBA, monomer and cross-linker). Being photosensitive, the polymerized microparticles progressively change their color when subjected to UV-vis irradiation. Above the LCST of the copolymer, the microparticles exhibited volume shrinkage accompanied by color deterioration. In addition, the UV light-driven clustering of the PNIPA-SPO copolymer was observed within the W/O droplet in the absence of photoinitiator, which contributed to variable microstructures from Janus to acorn-like and snowman-like morphologies. This work is the first attempt to unveil the photocontrolled asymmetric particle morphology by using the photoresponsive polymer.

Continuous sorting and separation of microparticles by size using AC dielectrophoresis in a PDMS microfluidic device with 3‐D conducting PDMS composite electrodes

Soft lithography technology allows for the development of numerous PDMS-based microfluidic devices for manipulation of particles and cells. However, integrating metallic electrodes with PDMS-based channel structures is challenging due to weak adhesion between metal and PDMS. To overcome this issue, we develop a new PDMS-based microfluidic device for continuous sorting and separation of microparticles by size using AC dielectrophoresis (DEP) with 3-D conducting PDMS composites as sidewall electrodes. The composites are synthesized by mixing silver powders with PDMS gel and such composite electrodes can easily be integrated with the PDMS microchannels. Furthermore, the sidewall electrodes also allow DEP forces to distribute three dimensionally, thus enhancing DEP effects in the entire region of channels. The capability of such PDMS-based microfluidic device is demonstrated for continuously sorting and separating 10 and 15 mum particles, and also for separating 5 from 10 mum particles. Together with experimental results, analysis of particle's trajectory based on Lagrangian approach provides insights into how microparticles transport under the effects of hydrodynamic and DEP forces in the present PDMS-based microfluidic device.

Preparation of Monodisperse Block Copolymer Vesicles via Flow Focusing in Microfluidics

We demonstrate that microfluidic flow devices enable a rapid, continuous, well-reproducible and size-controlled preparation of unilamellar block copolymer vesicles. The PDMS-based microfluidic device consists of perpendicularly crossed channels allowing hydrodynamic flow focusing of an ethanolic block copolymer solution in a stream of water. By altering the flow rate ratio in the water and ethanolic inlet channels, the vesicle size can be tuned over a large size range from 40 nm to 2 microm without subsequent processing steps manipulating size and shell characteristics. The ability of tuning the vesicle mean size over a range of several orders of magnitude with the possibility of in situ encapsulation of active ingredients creates new opportunities for the preparation of tailored drug delivery systems in science, medicine and industry.

Chemical control of Vorticella bioactuator using microfluidics

In this report, we demonstrate a microfluidic platform to control the stalk contraction and extension of Vorticella convallaria by changing concentration of Ca2+ with pneumatically-actuated elastomeric microvalves. Habitation, extraction and control of V. convallaria were carried out in a PDMS-based microfluidic device. By treating the cells with the permeant saponin, external actuation of cell-anchoring stalk between an extended and contracted state was achieved by cyclic exposure of the cells to a Ca2+ buffer (10(-6) M) and a rinse buffer containing EGTA as a chelation agent. When solutions were switched, the stalk contracted and extended responding to the ambient Ca2+ concentration change. The length of the stalk changed between 20 and 60 microm, resulting in a working distance of about 40 microm.

Recent developments in PDMS surface modification for microfluidic devices

PDMS is enjoying continued and ever increasing popularity as the material of choice for microfluidic devices due to its low cost, ease of fabrication, oxygen permeability and optical transparency. However, PDMS's hydrophobicity and fast hydrophobic recovery after surface hydrophilization, attributed to its low glass transition temperature of less than -120 degrees C, negatively impacts on the performance of PDMS-based microfluidic device components. This issue has spawned a flurry of research to devise longer lasting surface modifications of PDMS, with particular emphasis on microfluidic applications. This review will present recent research on surface modifications of PDMS using techniques ranging from metal layer coatings and layer-by-layer depositions to dynamic surfactant treatments and the adsorption of amphipathic proteins. We will also discuss significant advances that have been made with a broad palette of gas-phase processing methods including plasma processing, sol-gel coatings and chemical vapor deposition. Finally, we will present examples of applications and future prospects of modified PDMS surfaces in microfluidics, in areas such as molecular separations, cell culture in microchannels and biomolecular detection via immunoassays.

Detecting cytokine release from single T-cells.

The cytokine production by leukocytes correlates with body's ability to mount an immune response and therefore has high diagnostic value. In the present study we employed microfabricated surfaces to capture T-cells from minimally processed human blood, arrange these cells into a single cell array, and then detect interferon (IFN)-gamma released from individual cells. The fabrication of cell capture surfaces started with coating a silane-modified glass slide with a uniform layer of poly(ethylene glycol) (PEG) hydrogel. The hydrogel-coated slide was lyophilized and then incubated with a mixture of monoclonal anti-IFN-gamma and anti-CD4 antibodies (Abs). To define sites for single cell attachment, PEG hydrogel microwells (20 microm diameter) were photolithographically patterned on top of the Ab-containing hydrogel layer. This micropatterning process resulted in fabrication of PEG hydrogel microwells with Ab-decorated bottom and nonfouling walls. To minimize the blood volume requirement and to precisely define shear stress conditions, the engineered surface was enclosed inside a PDMS-based microfluidic device. Introduction of red blood cell (RBC) depleted whole human blood followed by controlled washing led to the isolation of individual CD4 T-cells within PEG microwells. Mitogenic activation and immunofluorescent staining performed inside the microfluidic chamber revealed IFN-gamma cytokine signal colocalized with specific T-cells. The device and process presented here will be expanded in the future to enable multiparametric functional analysis of immune cells organized into high density single cell arrays.

Vascular mimetics based on microfluidics for imaging the leukocyte–endothelial inflammatory response

We describe the development, validation, and application of a novel PDMS-based microfluidic device for imaging leukocyte interaction with a biological substrate at defined shear force employing a parallel plate geometry that optimizes experimental throughput while decreasing reagent consumption. The device is vacuum bonded above a standard 6-well tissue culture plate that accommodates a monolayer of endothelial cells, thereby providing a channel to directly observe the kinetics of leukocyte adhesion under defined shear flow. Computational fluid dynamics (CFD) was applied to model the shear stress and the trajectory of leukocytes within the flow channels at a micron length scale. In order to test this model, neutrophil capture, rolling, and deceleration to arrest as a function of time and position was imaged in the transparent channels. Neutrophil recruitment to the substrate proved to be highly sensitive to disturbances in flow streamlines, which enhanced the rate of neutrophil-surface collisions at the entrance to the channels. Downstream from these disturbances, the relationship between receptor mediated deceleration of rolling neutrophils and dose response of stimulation by the chemokine IL-8 was found to provide a functional readout of integrin activation. This microfluidic technique allows detailed kinetic studies of cell adhesion and reveals neutrophil activation within seconds to chemotactic molecules at concentrations in the picoMolar range.

Study of on-line monitoring of lactate based on optical fibre sensor and in-channel mixing mechanism

A PDMS-based microfluidic device with integrated optical fibres was developed for online monitoring of lactate. The detection is based on the optical adsorption of the colour solution formed by in-channel mixing of sample and reagent solutions. Computational Fluid Dynamics (CFD) simulation and kinetic study of the colour development were conducted to determine the minimum channel length and optimum residential time for homogenous mixing and maximum optical adsorption respectively. The system was proved to be able to detect lactate with good linearity (R(2)=0.98), response time of about 130 s and estimated limitation of detection (LOD) of 0.52 mM (about 47 mg/L), which is sufficient for a general online lactate detection. Due to the miniaturization of sensing system and micro-scale mixing, higher detection sensitivity (0.15 V/mM) can be realized than the mixing in lab-scale equipments (0.08 V/mM).

Circumventing air bubbles in microfluidic systems and quantitative continuous-flow PCR applications

Polymerase chain reaction (PCR) is an essential part of research based on genomics or cell analysis. The development of a microfluidic device that would be suitable for high-temperature-based reactions therefore becomes an important contribution towards the integration of micro-total analysis systems (muTAS). However, problems associated with the generation of air bubbles in the microchannels before the introduction of the assay liquid, which we call the "initial start-up" in this study, made the flow irregular and unstable. In this report, we have tried to address these problems by adapting a novel liquid-flow method for high-temperature-based reactions. A PDMS-based microfluidic device was fabricated by soft-lithography techniques and placed on a cartridge heater. The generation of the air bubbles was prevented by introducing the fluorinated oil, an inert and highly viscous liquid, as the cap just before the introduction of the sample solutions into the microchannels. The technique was applied for continuous-flow PCR, which could perform PCR on-chip in a microfluidic system. For the evaluation of practical accuracy, plasmid DNA that serves as a reference molecule for the quantification of genetically modified (GM) maize was used as the template DNA for continuous-flow PCR. After PCR, the products were collected in a vial and analyzed by gel electrophoresis to confirm the accuracy of the results. Additionally, quantitative continuous-flow PCR was performed using TaqMan technology on our PCR device. A laser detection system was also used for the quantitative PCR method. We observed a linear relationship between the threshold cycle (Ct) and the initial DNA concentration. These results showed that it would be possible to quantify the initial copies of the template DNA on our microfluidic device. Accurate quantitative DNA analysis in microfluidic systems is required for the integration of PCR with muTAS, thus we anticipate that our device would have promising potential for applications in a wide range of research.

Continuous-Flow Fractionation of Animal Cells in Microfluidic Device Using Aqueous Two-Phase Extraction

Monitoring of live cells is important in the field of medical science, diagnostics, biology, and the pharmaceutical industry. In this study, live and dead CHO-K1 (Chinese Hamster Ovary) cells were fractionated by continuous-flow extraction in a microfluidic device using immiscible aqueous two-phase extraction technique. The polymer solutions offered stable two-phase flows in microchannel without diffusive mixing. The fundamentals of aqueous two-phase extraction can support stable and reproducible recovery and separation of biomolecules in microfluidic devices. Polyethylene glycol 8000 (PEG 8000, 4%) and dextran T 500 (5%) were selected as model polymer solutions. The appropriate flow rates of polymer and cell solutions were suggested. The fractionation efficiency of live and dead CHO K-1 cells from the culture broth was compared in normal macroscale system and microfluidic device. The optimum pH for the fractionation was 6.6 in both the normal and micro-scale systems. The loss of target live cells by sedimentation was circumvented in microfluidic device because of the negligible effect of gravity on the sedimentation. Most live cells were distributed to PEG-rich phase, while dead cells were found at the interface of two polymer solutions in microchannel. In this case, the recovery and fractionation efficiency of live cells in the PDMS-based microfluidic device was 100% and 97.0%, respectively.

Chiral separation of dansyl amino acids by PDMS microchip gel monolithic column electrochromatography with γ-cyclodextrin bonded in polyacrylamide

A polydimethylsiloxane (PDMS) microchip polyacrylamide (PAA) gel monolithic column for electrochromatography was prepared. A chiral recognizable molecule, γ-cyclodextrin (γ-CD) was immobilized firmly in the PAA gel in the microchannel through chemical bond. The PDMS microchannel was pretreated with 3-(trimethoxysilyl)-propyl methacrylate (Bind-Silane) before bonding PAA monolithic column, which was an anchor molecule to connect the PAA gel with PDMS inner wall in microchannel. The experiments demonstrated γ-CD was a perfect chiral selector for fluorescein isoihiocyanate (FITC) labeled dansyl amino acids (Dns-AAs), which showed well capability for chiral recognization in monolithic column electrochromatography. The mixture of two pairs of FITC-labeled Dns-AAs was separated absolutely in this new-style PDMS-based microfluidic device monolithic column within 100 s in 36 mm effective separation length, and the resolutions of the two pairs of optical isomers could reach 2.53 and 1.12, respectively. The condition of preparation PAA gel monolithic column for chiral separation was optimized by adjusting the concentration of allyl-γ-CD. The stability, transferring heat and optical characteristic of PDMS-based microfluidic device monolithic column were examined, respectively.

PDMS-based microfluidic device with multi-height structures fabricated by single-step photolithography using printed circuit board as masters.

We have developed a method for fabricating microfluidic devices with multi-height structures using single step photolithography. The whole fabrication process is executed by conventional printed circuit board (PCB) technology without the need of having access to clean room facilities. Specifically designed "windows" and "rims" architectures were printed on films that were used as photomasks. Different levels of protruding features on the PCB master were produced by exposing a photomask followed by chemical wet etching. Poly(dimethylsiloxane) (PDMS) was then moulded against the positive relief master to generate microfluidic structures. In this report, we described the fabrication of a microfluidic device featured with a multi-height "sandbag" structure for particle entrapment and peripheral microchannels. Controlled immobilization of biological cells and immunocytochemcial staining assays were performed to demonstrate the applicability of the microfluidic device for cellular analysis. The integrity of the microdevice remained stable under applied pressure, indicating the robustness of the elastic PDMS structures for analytical operation. The simple microfabrication process requires only low-cost materials and minimal specialized equipment and can reproducibly produce mask lines of about 20 microm in width, which is sufficient for most microfluidic applications.