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.     

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.     

Implementation of tetra-poly(ethylene glycol) hydrogel with high mechanical strength into microfluidic device technology

Hydrogels have several excellent characteristics suitable for biomedical use such as softness, biological inertness and solute permeability. Hence, integrating hydrogels into microfluidic devices is a promising approach for providing additional functions such as biocompatibility and porosity, to microfluidic devices. However, the poor mechanical strength of hydrogels has severely limited device design and fabrication. A tetra-poly(ethylene glycol) (tetra-PEG) hydrogel synthesized recently has high mechanical strength and is expected to overcome such a limitation. In this research, we have comprehensively studied the implementation of tetra-PEG gel into microfluidic device technology. First, the fabrication of tetra-PEG gel/PDMS hybrid microchannels was established by developing a simple and robust bonding technique. Second, some fundamental features of tetra-PEG gel/PDMS hybrid microchannels, particularly fluid flow and mass transfer, were studied. Finally, to demonstrate the unique application of tetra-PEG-gel-integrated microfluidic devices, the generation of patterned chemical modulation with the maximum concentration gradient: 10% per 20 μm in a hydrogel was performed. The techniques developed in this study are expected to provide fundamental and beneficial methods of developing various microfluidic devices for life science and biomedical applications.     

Microfluidic electrospinning of biphasic nanofibers with Janus morphology

In this paper a method of electrospinning conducting and nonconducting biphasic Janus nanofibers using microfluidic polydimethylsiloxane (PDMS)-based manifolds is described. Key benefits of using microfluidic devices for nanofiber synthesis include rapid prototyping, ease of fabrication, and the ability to spin multiple Janus fibers in parallel through arrays of individual microchannels. Biphasic Janus nanofibers of polyvinylpyrrolidone (PVP)+polypyrrole (PPy)∕PVP nanofibers with an average diameter of 250 nm were successfully fabricated using elastomeric microfluidic devices. Fiber characterization and confirmation of the Janus morphology was subsequently carried out using a combination of scanning electron microscopy, energy dispersion spectroscopy, and transmission electron microscopy.     

Deformation properties between fluid and periodic circular obstacles in polydimethylsiloxane microchannels: Experimental and numerical investigations under various conditions

Understanding the mechanical properties of optically transparent polydimethylsiloxane (PDMS) microchannels was essential to the design of polymer-based microdevices. In this experiment, PDMS microchannels were filled with a 100 μM solution of rhodamine 6G dye at very low Reynolds numbers (∼10⁻³). The deformation of PDMS microchannels created by pressure-driven flow was investigated by fluorescence microscopy and quantified the deformation by the linear relationship between dye layer thickness and intensity. A line scan across the channel determined the microchannel deformation at several channel positions. Scaling analysis widely used to justify PDMS bulging approximation was allowed when the applied flow rate was as high as 2.0 μl/min. The three physical parameters (i.e., flow rate, PDMS wall thickness, and mixing ratio) and the design parameter (i.e., channel aspect ratio = channel height/channel width) were considered as critical parameters and provided the different features of pressure distributions within polymer-based microchannel devices. The investigations of the four parameters performed on flexible materials were carried out by comparison of experiment and finite element method (FEM) results. The measured Young''s modulus from PDMS tensile test specimens at various circumstances provided reliable results for the finite element method. A thin channel wall, less cross-linker, high flow rate, and low aspect ratio microchannel were inclined to have a significant PDMS bulging. Among them, various mixing ratios related to material property and aspect ratios were one of the significant factors to determine PDMS bulging properties. The measured deformations were larger than the numerical simulation but were within corresponding values predicted by the finite element method in most cases.     

One-heater flow-through polymerase chain reaction device by heat pipes cooling

This study describes a novel microfluidic reactor capable of flow-through polymerase chain reactions (PCR). For one-heater PCR devices in previous studies, comprehensive simulations and experiments for the chip geometry and the heater arrangement were usually needed before the fabrication of the device. In order to improve the flexibility of the one-heater PCR device, two heat pipes with one fan are used to create the requisite temperature regions in our device. With the integration of one heater onto the chip, the high temperature required for the denaturation stage can be generated at the chip center. By arranging the heat pipes on the opposite sides of the chip, the low temperature needed for the annealing stage is easy to regulate. Numerical calculations and thermal measurements have shown that the temperature distribution in the five-temperature-region PCR chip would be suitable for DNA amplification. In order to ensure temperature uniformity at specific reaction regions, the Re of the sample flow is less than 1. When the microchannel width increases and then decreases gradually between the denaturation and annealing regions, the extension region located in the enlarged part of the channel can be observed numerically and experimentally. From the simulations, the residence time at the extension region with the enlarged channel is 4.25 times longer than that without an enlarged channel at a flow rate of 2 μl/min. The treated surfaces of the flow-through microchannel are characterized using the water contact angle, while the effects of the hydrophilicity of the treated polydimethylsiloxane (PDMS) microchannels on PCR efficiency are determined using gel electrophoresis. By increasing the hydrophilicity of the channel surface after immersing the PDMS substrates into Tween 20 (20%) or BSA (1 mg/ml) solutions, efficient amplifications of DNA segments were proved to occur in our chip device. To our knowledge, our group is the first to introduce heat pipes into the cooling module that has been designed for a PCR device. The unique architecture utilized in this flow-through PCR device is well applied to a low-cost PCR system.     

Surface modification on polydimethylsiloxane-based microchannels with fragmented poly(l-lactic acid) nanosheets

Surface modification is a critical issue in various applications of polydimethylsiloxane (PDMS)-based microfluidic devices. Here, we describe a novel method through which PDMS-based microchannels were successfully modified with fragmented poly(l-lactic acid) (PLLA) nanosheets through a simple patchwork technique that exploited the high level of adhesiveness of PLLA nanosheets. Compared with other surface modification methods, our method required neither complicated chemical modifications nor the use of organic solvents that tend to cause PDMS swelling. The experimental results indicated that the modified PDMS exhibited excellent capacity for preventing the adhesion and activation of platelets. This simple yet efficient method can be used to fabricate the special PDMS microfluidic devices for biological, medical, and even hematological purposes.     

In situ pressure measurement within deformable rectangular polydimethylsiloxane microfluidic devices

In this paper, we present a simple procedure to incorporate commercially available external pressure transducers into existing microfluidic devices, to monitor pressure-drop in real-time, with minimal design modifications to pre-existing channel designs. We focus on the detailed fabrication steps and assembly to make the process straightforward and robust. The work presented here will benefit those interested in adding pressure drop measurements in polydimethylsiloxane (PDMS) based microchannels without having to modify existing channel designs or requiring additional fabrication steps. By using three different devices with varying aspect ratio channels (wh0, width/depth), we demonstrate that our approach can easily be adapted into existing channel designs inexpensively. Furthermore, our approach can achieve steady state measurements within a matter of minutes (depending on the fluid) and can easily be used to investigate dynamic pressure drops. In order to validate the accuracy of the measured pressure drops within the three different aspect ratio devices, we compared measured pressure drops of de-ionized water and a 50 wt. % glycerol aqueous solution to four different theoretical expressions. Due to the deformability of PDMS, measured pressure drops were smaller than those predicted by the rigid channel theories (plate and rectangular). Modification of the rigid channel theories with a deformability parameter α provided better fits to the measured data. The elastic rectangular expression developed in this paper does not have a geometric restriction and is better suited for microchannels with a wider range of aspect ratios.     

Detection of immunoglobulins in a laser induced fluorescence system utilizing polydimethysiloxane microchips with advanced surface and optical properties

We developed an automated laser induced fluorescence system utilizing microfluidic chips for detection and quantification of immunoglobulins. Microchips were fabricated from polydimethysiloxane (PDMS) using the so-called "prepolymerization technique." The microchip structure helped minimize the effects of PDMS autofluorescence and light scattering. Furthermore, a thin and uniform PDMS layer forming the top of the microchip enabled proper focusing and collection of the excitation beam and the emitted fluorescence, respectively. The developed system was tested for the detection of mouse immunoglobulins. The capturing antibodies were immobilized on internal microchannel walls in the form of a polyelectrolyte. We clearly show that this immobilization technique, if correctly realized, gives results with high reproducibility. After sample incubation and washing, secondary antibodies labeled by fluorescein isothiocyanate were introduced into microchannels to build a detectable complex. We show that mouse antibodies can be quantified in a wide concentration range, 0.01-100 μg ml(-1). The lower detection limit was below 0.001 μg ml(-1) (6.7 pM). The developed laser induced fluorescence (LIF) apparatus is relatively cheap and easy to construct. The total cost of the developed LIF detector is lower than a typical price of plate readers. If compared to classical ELISA (enzyme linked immunosorbent assay) plate systems, the detection of immunoglobulins or other proteins in the developed PDMS microfluidic device brings other important benefits such as reduced time demands (10 min incubation) and low reagent consumption (less than 1 μl). The cost of the developed PDMS chips is comparable with the price of commercial ELISA plates. The main troubleshooting related to the apparatus development is also discussed in order to help potential constructors.     

A simple method of fabricating mask-free microfluidic devices for biological analysis

We report a simple, low-cost, rapid, and mask-free method to fabricate two-dimensional (2D) and three-dimensional (3D) microfluidic chip for biological analysis researches. In this fabrication process, a laser system is used to cut through paper to form intricate patterns and differently configured channels for specific purposes. Bonded with cyanoacrylate-based resin, the prepared paper sheet is sandwiched between glass slides (hydrophilic) or polymer-based plates (hydrophobic) to obtain a multilayer structure. In order to examine the chip’s biocompatibility and applicability, protein concentration was measured while DNA capillary electrophoresis was carried out, and both of them show positive results. With the utilization of direct laser cutting and one-step gas-sacrificing techniques, the whole fabrication processes for complicated 2D and 3D microfluidic devices are shorten into several minutes which make it a good alternative of poly(dimethylsiloxane) microfluidic chips used in biological analysis researches.     

Direct detection of cancer biomarkers in blood using a “place n play” modular polydimethylsiloxane pump

Cancer biomarkers have significant potential as reliable tools for the early detection of the disease and for monitoring its recurrence. However, most current methods for biomarker detection have technical difficulties (such as sample preparation and specific detector requirements) which limit their application in point of care diagnostics. We developed an extremely simple, power-free microfluidic system for direct detection of cancer biomarkers in microliter volumes of whole blood. CEA and CYFRA21-1 were chosen as model cancer biomarkers. The system automatically extracted blood plasma from less than 3 μl of whole blood and performed a multiplex sample-to-answer assay (nano-ELISA (enzyme-linked immunosorbent assay) technique) without the use of external power or extra components. By taking advantage of the nano-ELISA technique, this microfluidic system detected CEA at a concentration of 50 pg/ml and CYFRA21-1 at a concentration of 60 pg/ml within 60 min. The combination of PnP polydimethylsiloxane (PDMS) pump and nano-ELISA technique in a single microchip system shows great promise for the detection of cancer biomarkers in a drop of blood.     

A pneumatic valve controlled microdevice for bioanalysis

This paper describes a pneumatic valve controlled microdevice for performing mixing and reaction. This microdevice combined the degassed polydimethylsiloxane (PDMS) pumping method with a syringe-actuated valve system to control the dispensing and mixing of nanoliter solutions. The syringe was used to manually generate vacuum and to open the valves. Upon the opening of the valve, the microchamber was filled with the solution, which was driven by the external atmosphere through the degassed PDMS microchannel. With this microdevice, the enzymatic kinetics of alkaline phosphatase converting the fluorescein diphosphate was studied, and the Michaelis-Menten kinetics was analyzed. The microdevice has the advantages of simplicity and low cost in fabrication and operation.     

Polydimethyl siloxane wet etching for three dimensional fabrication of microneedle array and high-aspect-ratio micropillars

Among various transdermal drug delivery (TDD) approaches, utilizing the microneedles (MNs) not only can penetrate the skin but also deliver the drug with reduced tissue damage, reduced pain, and no bleeding. However, the MNs with larger height are required to overcome the skin barrier for effective TDD. Unlike 2D patterning, etching polydimethyl siloxane (PDMS) micropillars for fabrication of 3D microstructures is presented. The PDMS micropillars were first constructed by casting PDMS on the computer numerical control-machined cylindrical microwells, which then went through etching process to obtain the MNs for subsequent fabrication of polymer MNs or high aspect ratio micropillars.     

Reduction of water evaporation in polymerase chain reaction microfluidic devices based on oscillating-flow

Producing polymeric or hybrid microfluidic devices operating at high temperatures with reduced or no water evaporation is a challenge for many on-chip applications including polymerase chain reaction (PCR). We study sample evaporation in polymeric and hybrid devices, realized by glass microchannels for avoiding water diffusion toward the elastomer used for chip fabrication. The method dramatically reduces water evaporation in PCR devices that are found to exhibit optimal stability and effective operation under oscillating-flow. This approach maintains the flexibility, ease of fabrication, and low cost of disposable chips, and can be extended to other high-temperature microfluidic biochemical reactors.     

High yield fabrication of multilayer polydimethylsiloxane devices with freestanding micropillar arrays

A versatile method to fabricate a multilayer polydimethylsiloxane (PDMS) device with micropillar arrays within the inner layer is reported. The method includes an inexpensive but repeatable approach for PDMS lamination at high compressive force to achieve high yield of pillar molding and transfer to a temporary carrier. The process also enables micropillar-containing thin films to be used as the inner layer of PDMS devices integrated with polymer membranes. A microfluidic cell culture device was demonstrated which included multiple vertically stacked flow channels and a pillar array serving as a cage for a collagen hydrogel. The functionality of the multilayer device was demonstrated by culturing collagen-embedded fibroblasts under interstitial flow through the three-dimensional scaffold. The fabrication methods described in this paper can find applications in a variety of devices, particularly for organ-on-chip applications.     

An off-the-shelf integrated microfluidic device comprising self-assembled monolayers for protein array experiments

Microfluidic-based protein arrays are promising tools for life sciences, with increased sensitivity and specificity. One of the drawbacks of this technology is the need to create fresh surface chemistry for protein immobilization at the beginning of each experiment. In this work, we attempted to include the process of surface functionalization as part of the fabrication of the device, which would substitute the time consuming step of surface functionalization at the beginning of each protein array experiment. To this end, we employed a novel surface modification using self-assembled monolayers (SAMs) to immobilize biomolecules within the channels of a polydimethylsiloxane (PDMS) integrated microfluidic device. As a model, we present a general method for depositing siloxane-anchored SAMs, with 1-undecyl-thioacetate-trichlorosilane (C₁₁TA) on the silica surfaces. The process involved developing PDMS-compatible conditions for both SAM deposition and functional group activation. We successfully demonstrated the ability to produce, within an integrated microfluidic channel, a C₁₁TA monolayer with a covalently conjugated antibody. The antibody could then bind its antigen with a high signal to background ratio. We further demonstrated that the antibody was still active after storage of the device for a week. Integration of the surface chemistry into the device as part of its fabrication process has potential to significantly simplify and shorten many experimental procedures involving microfluidic–based protein arrays. In turn, this will allow for broader dissemination of this important technology.     

A new fabrication technique to form complex polymethylmethacrylate microchannel for bioseparation

Recent studies show that reduction in cross-sectional area can be used to improve the concentration factor in microscale bioseparations. Due to simplicity in fabrication process, a step reduction in cross-sectional area is generally implemented in microchip to increase the concentration factor. But the sudden change in cross-sectional area can introduce significant band dispersion and distortion. This paper reports a new fabrication technique to form a gradual reduction in cross-sectional area in polymethylmethacrylate (PMMA) microchannel for both anionic and cationic isotachophoresis (ITP). The fabrication technique is based on hot embossing and surface modification assisted bonding method. Both one-dimensional and two-dimensional gradual reduction in cross-sectional area microchannels were formed on PMMA with high fidelity using proposed techniques. ITP experiments were conducted to separate and preconcentrate fluorescent proteins in these microchips. Thousand fold and ten thousand fold increase in concentrations were obtained when 10 × and 100 × gradual reduction in cross-sectional area microchannels were used for ITP.     

Microfluidic carbon-blackened polydimethylsiloxane device with reduced ultra violet background fluorescence for simultaneous two-color ultra violet/visible-laser induced fluorescence detection in single cell analysis

In single cell analysis (SCA), individual cell-specific properties and inhomogeneous cellular responses are being investigated that is not subjected to ensemble-averaging or heterogeneous cell population effects. For proteomic single cell analysis, ultra-sensitive and reproducible separation and detection techniques are essential. Microfluidic devices combined with UV laser induced fluorescence (UV-LIF) detection have been proposed to fulfill these requirements. Here, we report on a novel microfluidic chip fabrication procedure that combines straightforward production of polydimethylsiloxane (PDMS) chips with a reduced UV fluorescence background (83%-reduction) by using PDMS droplets with carbon black pigments (CBP) as additives. The CBP-droplet is placed at the point of detection, whereas the rest of the chip remains transparent, ensuring full optical control of the chip. We systematically studied the relation of the UV background fluorescence at CBP to PDMS ratios (varying from 1:10 to 1:1000) for different UV laser powers. Using a CBP/PDMS ratio of 1:20, detection of a 100 nM tryptophan solution (S/N = 3.5) was possible, providing a theoretical limit of detection of 86 nM (with S/N = 3). Via simultaneous two color UV/VIS-LIF detection, we were able to demonstrate the electrophoretic separation of an analyte mixture of 500 nM tryptophan (UV) and 5 nM fluorescein (VIS) within 30 s. As an application, two color LIF detection was also used for the electrophoretic separation of the protein content from a GFP-labeled single Spodoptera frugiperda (Sf9) insect cell. Thereby just one single peak could be measured in the visible spectral range that could be correlated with one single peak among others in the ultraviolet spectra. This indicates an identification of the labeled protein γ-PKC and envisions a further feasible identification of more than one single protein in the future.     

A large-area hemispherical perforated bead microarray for monitoring bead based aptamer and target protein interaction

Herein, we present a large-area 3D hemispherical perforated microwell structure for a bead based bioassay. Such a unique microstructure enables us to perform the rapid and stable localization of the beads at the single bead level and the facile manipulation of the bead capture and retrieval with high speed and efficiency. The fabrication process mainly consisted of three steps: the convex micropatterned nickel (Ni) mold production from the concave micropatterned silicon (Si) wafer, hot embossing on the polymer matrix to generate the concave micropattened acrylate sheet, and reactive ion etching to make the bottom holes. The large-area hemispherical perforated micropatterned acrylate sheet was sandwiched between two polydimethylsiloxane (PDMS) microchannel layers. The bead solution was injected and recovered in the top PDMS microchannel, while the bottom PDMS microchannel was connected with control lines to exert the hydrodynamic force in order to alter the flow direction of the bead solution for the bead capture and release operation. The streptavidin-coated microbead capture was achieved with almost 100% yield within 1 min, and all the beads were retrieved in 10 s. Lysozyme or thrombin binding aptamer labelled microbeads were trapped on the proposed bead microarray, and the in situ fluorescence signal of the bead array was monitored after aptamer-target protein interaction. The protein-aptamer conjugated microbeads were recovered, and the aptamer was isolated for matrix assisted laser desorption/ionization time-of-flight mass spectrometry analysis to confirm the identity of the aptamer.     

Evaluation of 3D-printed molds for fabrication of non-planar microchannels

Replica obtained from micromolds patterned by simple photolithography has features with uniform heights, and attainable microchannels are thus quasi-two-dimensional. Recent progress in three-dimensional (3D) printing has enabled facile desktop fabrication of molds to replicate microchannels with varying heights. We investigated the replica obtained from four common techniques of 3D printing—fused deposition modeling, selective laser sintering, photo-polymer inkjet printing (PJ), and stereolithography (SL)—for the suitability to form microchannels in terms of the surface roughness inherent to the mechanism of 3D printing. There have been limited quantitative studies that focused on the surface roughness of a 3D-printed mold with different methods of 3D printing. We discussed that the surface roughness of the molds affected (1) transparency of the replica and (2) delamination pressure of poly(dimethylsiloxane) replica bonded to flat glass substrates. Thereafter, we quantified the accuracy of replication from 3D-printed molds by comparing the dimensions of the replicated parts to the designed dimensions and tested the ability to fabricate closely spaced microchannels. This study suggested that molds printed by PJ and SL printers were suitable for replica molding to fabricate microchannels with varying heights. The insight from this study shall be useful to fabricate 3D microchannels with controlled 3D patterns of flows guided by the geometry of the microchannels.