EWOD microfluidic systems for biomedical applications

As the technology advances, a growing number of biomedical microelectromechanical systems (bio-MEMS) research involves development of lab-on-a-chip devices and micrototal analysis systems. For example, a portable instrument capable of biomedical analyses (e.g., blood sample analysis) and immediate recording, whether the patients are in the hospital or home, would be a considerable benefit to human health with an excellent commercial viability. Digital microfluidic (DMF) system based on the electrowetting-on-dielectric (EWOD) mechanism is an especially promising candidate for such point-of-care systems. The EWOD-based DMF system processes droplets in a thin space or on an open surface, unlike the usual microfluidic systems that process liquids by pumping them in microchannels. Droplets can be generated and manipulated on EWOD chip only with electric signals without the use of pumps or valves, simplifying the chip fabrication and the system construction. Microfluidic operations by EWOD actuation feature precise droplet actuation, less contamination risk, reduced reagents volume, better reagents mixing efficiency, shorter reaction time, and flexibility for integration with other elements. In addition, the simplicity and portability make the EWOD-based DMF system widely popular in biomedical or chemical fields as a powerful sample preparation platform. Many chemical and biomedical researches, such as DNA assays, proteomics, cell assays, and immunoassays, have been reported using the technology. In this paper, we have reviewed the recent developments and studies of EWOD-based DMF systems for biomedical applications published mostly during the last 5 years.     

Controllable fission of droplets and bubbles by pneumatic valve

We report a feasible method that can precisely control the fission of droplets by modulating the flow resistance using pneumatic valves. Multilayer soft lithography was used to fabricate the valves. They can be used as variable microfluidic resistor (VMR) to dramatically change the flow resistance. A simulation has been done to forecast the behavior of droplets. We used this technique to control break-up of generated droplets and direct their motion. Droplets with different volume ratios were obtained in one chip. To investigate the mechanism, an equivalent electrical circuit was introduced to compare with the fluid network. This method could potentially be applied to different geometries, especially for the microfluidic network consisting of a set of two parallel channels with a common inlet and different outlets in bifurcating channels. Besides, manipulation of bubbles was also demonstrated.     

Highly integrated polymeric microliquid flow controller for droplet microfluidics

Microfluidic applications demand accurate control and measurement of small fluid flows and volumes, and the majority of approaches found in the literature involve materials and fabrication methods not suitable for a monolithic integration of different microcomponents needed to make a complex Lab-on-a-Chip (LoC) system. The present work leads to a design and manufacturing approach for problem-free monolithic integration of components on thermoplastics, allowing the production of excellent quality devices either as stand-alone components or combined in a complex structures. In particular, a polymeric liquid flow controlling system (LFCS) at microscale is presented, which is composed of a pneumatic microvalve and an on-chip microflow sensor. It enables flow regulation between 30 and 230 μl/min with excellent reproducibility and accuracy (error lower than 5%). The device is made of a single Cyclic Olefin Polymer (COP) piece, where the channels and cavities are hot-embossed, sealed with a single COP membrane by solvent bonding and metalized, after sealing, to render a fully functional microfluidic control system that features on-chip flow sensing. In contrast with commercially available flow control systems, the device can be used for high-quality flow modulation in disposable LoC devices, since the microfluidic chip is low cost and replaceable from the external electronic and pneumatic actuators box. Functionality of the LFCS is tested by connecting it to a microfluidic droplet generator, rendering highly stable flow rates and allowing generation of monodisperse droplets over a wide range of flow rates. The results indicate the successful performance of the LFCS with significant improvements over existing LFCS devices, facing the possibility of using the system for biological applications such as generating distinct perfusion modes in cell culture, novel digital microfluidics. Moreover, the integration capabilities and the reproducible fabrication method enable straightforward transition from prototype to product in a way that is lean, cost-effective and with reduced risk.     

3D-printed membrane microvalves and microdecoder

A microfluidic system for multichannel switching and multiphase flow control has potential uses in pneumatic soft robotics and biological sampling systems. At present, the membrane microvalves used in microfluidic systems are mostly constructed using a multilayer bonding process so that the device cannot withstand high pressures. In this paper, we demonstrate a design method and the properties of a bondless membrane microvalve fabricated using a commercial 3D printer. We used a multijet (MJP) 3D printer to print a 100-μm-thick and 6-mm-diameter membrane from a relatively hard material (1700 MPa). The membrane’s high toughness ensures that it does not need negative pressure to reopen. The measured operation frequency was less than 2.5 Hz under a pneumatic pressure of 14.5 kPa. We also 3D-printed an integrated Quake-style microfluidic decoder network by combining 8 valves in series to demonstrate the integrability of the microvalve. The decoder chip was demonstrated to control the ON/OFF state of the four coded fluidic channels, with the droplets being generated from selected channels according to the valve action. Therefore, such 3D-printed microvalves are highly integrable, have a high manufacturing efficiency, and can be applied in pneumatic controllers, sample switchers and integrated print heads.

     

Abstraction layers for scalable microfluidic biocomputing

Microfluidic devices are emerging as an attractive technology for automatically orchestrating the reactions needed in a biological computer. Thousands of microfluidic primitives have already been integrated on a single chip, and recent trends indicate that the hardware complexity is increasing at rates comparable to Moore’s Law. As in the case of silicon, it will be critical to develop abstraction layers—such as programming languages and Instruction Set Architectures (ISAs)—that decouple software development from changes in the underlying device technology. Towards this end, this paper presents BioStream, a portable language for describing biology protocols, and the Fluidic ISA, a stable interface for microfluidic chip designers. A novel algorithm translates microfluidic mixing operations from the BioStream layer to the Fluidic ISA. To demonstrate the benefits of these abstraction layers, we build two microfluidic chips that can both execute BioStream code despite significant differences at the device level. We consider this to be an important step towards building scalable biological computers.

Printing 3D microfluidic chips with a 3D sugar printer

This study demonstrated how to quickly and effectively print two-dimensional (2D) and three-dimensional (3D) microfluidic chips with a low-cost 3D sugar printer. The sugar printer was modified from a desktop 3D printer by redesigning the extruder, so the melting sugar could be extruded with pneumatic driving. Sacrificial sugar lines were first printed on a base layer followed by casting polydimethylsiloxane (PDMS) onto the layer and repeating. Microchannels were then printed in the PDMS solvent, microfluidic chips dropped into hot water to dissolve the sugar lines after the PDMS was solidified, and the microfluidic chips did not need further sealing. Different types of sugar utilized for printing material were studied with results indicating that maltitol exhibited a stable flow property compared with other sugars such as caramel or sucrose. Low cost is a significant advantage of this type of sugar printer as the machine may be purchased for only approximately $800. Additionally, as demonstrated in this study, the printed 3D microfluidic chip is a useful tool utilized for cell culture, thus proving the 3D printer is a powerful tool for medical/biological research.     

Inhibition of on-chip PCR using PDMS–glass hybrid microfluidic chips

In the course of developing a microfluidic analytical platform incorporating the polymerase chain reaction (PCR) and subsequent capillary electrophoresis (CE) analysis for a variety of bio-assays, we examined PCR inhibition through surface interactions with the chip materials. Our devices perform PCR in a three-layer chip, a glass–poly(dimethylsiloxane)–glass sandwich in which the poly(dimethylsiloxane) (PDMS, a silicone rubber) layer is used for pneumatic membrane pumping and valving of the PCR reagents. Initial on-chip PCR–CE tests of BK virus replicated in multiple uncoated chips showed variable results, usually yielding no detectable product at the target sample concentrations used. Subsequent “chip-flush” experiments, where water or reagents were flushed through a chip and subsequently incorporated in off-chip PCR, highlighted bovine serum albumin (BSA) amongst other pre-treatments, chip materials and PCR recipes as being effective in mitigating inhibition. When the BSA channel pre-coating was applied to on-chip PCR–CE experiments, a substantial improvement (10× to 40×) in signal-to-noise (S/N) of the CE product peak was conferred, and was shown with high confidence despite high S/N variability. This is the first study to quantitatively examine BSA’s ability to reduce inhibition of PCR performed on PDMS chips, and one of very few microfluidic PCR inhibition studies of any kind to use a large number of microfluidic chips (~400). The simplicity and effectiveness of our BSA coating suggest that passivating materials applied to microfluidic device channel networks may provide a viable pathway for development of bio-compatible devices with reduced complexity and cost.     

Monolithic fabrication of electro-fluidic polymer microchips

Integration of electronic wiring with microfluidic chips is an important process as it allows electrical interactions with the fluidic media, for example required for resistive and capacitive sensing. It is also necessary in order to implement various actuation and control mechanisms such as pumping, electrophoresis and temperature control. Typically electrical wire traces are added to microfabricated fluidic chips using metal deposition processes that are carried out after the fluidic chip has been fabricated. The process for adding the wiring is complicated and is limited to select metals that can be deposited by evaporation or sputtering. We present a single step method for integrating electrical wires into polymer microfluidic chips that are fabricated by a hot embossing process. This process can flexibly embed any kind of commercially available metal wire with a microfluidic chip and the wiring may be integrated to come into surface contact with the fluid or may be embedded in close proximity to (but insulated from)the fluid paths for example for local heating purposes. This method significantly reduces total processing time and is thus a valuable method for wire integration into polymer chips. We demonstrate two applications—a microelectrolysis chip and a heater chip that were fabricated using this methodology. The design, fabrication process and the initial test results are presented.     

Manufacturing and packaging of sensors for their integration in avertical MCM microsystem for biomedical applications

Demonstrates the feasibility of integrating fragile micromachined chips into a complex three-dimensional (3-D) multichip module (MCM) microsystem for a biomedical application. The system is based on the vertical integration of the different parts: micropumps and valves, a multisensor chip for on-line control of the system and a signal-processing chip. In this paper, packaging of the microsystem is studied in order to minimize the induced stress that can affect the integrity of the different micromachined parts of the system. Standard commercially available components and materials were used so as to minimize costs for the case of high volume packaging. For testing the approach, a multisensor chip which includes thin silicon membrane-based devices has been used as the main test structure to compare different packaging materials. In addition, for the fabrication of such a sensor chip in an efficient mode, technological modules needed to fabricate sensors on complementary metal-oxide-semiconductor (CMOS) wafers are discussed. The definition of standardized "add-on" sensor modules to the CMOS process of a foundry is intended to limit the development cost of smart sensors     

Detection of Cellular Parameters Using a Microfluidic Chip-Based System

Lab-on-a-chip technology achieves a reduction of sample and reagent volume and automates complex laboratory processes. Here, we present the implementation of cell assays on a microfluidic platform using disposable microfluidic chips. The applications are based on the controlled movement of cells by pressure-driven flow inside networks of microfluidic channels. Cells are hydrodynamically focused and pass the fluorescence detector in single file. Initial applications are the determination of protein expression and apoptosis parameters. The microfluidic system allows unattended measurement of six samples per chip. Results obtained with the microfluidic chips showed good correlation with data obtained using a standard flow cytometer.     

Design of a flow-controlled asymmetric droplet splitter using computational fluid dynamics

In this work the design of a segmented flow microfluidic device is presented that allows droplet splitting ratios from 1:1 up to 20:1. This ratio can be dynamically changed on chip by altering an additional oil flow. The design was fabricated in PDMS chips using the standard SU-8 mold technique and does not require any valves, membranes, optics or electronics. To avoid a trial and error approach, fabricating and testing several designs, a computational fluid dynamics model was developed and validated for droplet formation and splitting. The model was used to choose between several variations of the splitting T-junction with the extra oil inlet, as well to predict the additional flow rate needed to split the droplets in various ratios. Experimental and simulated results were in line, suggesting the model’s suitability to optimize future designs and concepts. The resulting asymmetric droplet splitter design opens possibilities for controlled sampling and improved magnetic separation in bio-assay applications.     

Interfacing microfluidics to LDI-MS by automatic robotic spotting

We developed a method of interfacing microfluidics with mass spectrometry (MS) using a robotic spotting system to automate the contact spotting process. We demonstrate that direct and automated spotting of analyte from multichannel microfluidic chips to a custom microstructured MALDI target plate was a simple, robust, and high-throughput method for interfacing parallel microchannels using matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). Using thermoplastic cyclic olefin copolymer (COC) polymer microfluidic chips containing eight parallel 100 μm × 46 μm microchannels connected to a single input port, spotting volume repeatability and MALDI-MS signal uniformity are evaluated for a panel of sample peptides. The COC microfluidic chips were fabricated by hot embossing and solvent bonding techniques followed by chip dicing to create open ends for MS interfacing. Using the automatic robotic spotting approach, microfluidic chip-based reversed-phase liquid chromatography (RPLC) separations were interfaced with electrochemically etched nanofilament silicon (nSi) target substrate, demonstrating the potential of this approach toward chip-based microfluidic separation coupled with matrix-free laser desorption/ionization mass spectrometry.     

Integration of Microfluidics With a Love Wave Sensor for the Fabrication of a Multisample Analytical Microdevice

A novel approach for multisensing has been developed based on the integration of a parallel-channel microfluidic module with a surface acoustic wave (SAW) sensor chip. The microfluidic module was used to compartmentalize the surface of a single SAW sensor into $N$ equal subareas in order to deliver and detect multiple samples on the sensor. The design concerns and fabrication procedure using soft lithography of polydimethylsiloxane are described. Successful demonstration of a four-channel module is reported, along with a sensitivity evaluation and comparison with a standard flow cell used so far. Very promising results were revealed during the tests concerning the system's operation with liquid samples. The reliability and reproducibility of the results in all four subareas render the proposed setup very suitable for biological testing and screening of various biomolecules in an array format.$ hfill$[2008-0031]      

Determining refractive index of single living cell using an integrated microchip

We report a novel method for measuring the effective refractive index (RI) of single living cell using a small integrated chip, which might be an efficient approach for diseases diagnosis. This microchip is able to determine the refractive index of single living cell in real time without any extra cell treatments such as fluorescence labelling, chemical modification and so forth, meanwhile, providing low cost, small size, easy operation and high accuracy. The measurement system integrates an external cavity laser, a microlens, and some microfluidic channels onto a monolithic chip. In the experiments, two standard polystyrene beads with nominal RI are utilized to calibrate the measurement system and five different types of cancerous cells are subsequently measured in the chip. The experimental results show that the refractive indices of the cancerous cells tested ranges from 1.392 to 1.401, which is larger than typical value of normal cell of 1.35–1.37. This integrated chip potentially has a serial of applications on biodefense, disease diagnosis, biomedical and biochemical analysis.     

Rapid prototyping of PMMA microfluidic chips utilizing a CO<Subscript>2</Subscript> laser

A commercially available CO₂ laser scriber is used to perform the direct-writing ablation of polymethyl-methacrylate (PMMA) substrates for microfluidic applications. The microfluidic designs are created using commercial layout software and are converted into the command signals required to drive the laser scriber in such a way as to reproduce the desired microchannel configuration on the surface of a PMMA substrate. The aspect ratio and surface quality of the ablated microchannels are examined using scanning electron microscopy and atomic force microscopy surface measurement techniques. The results show that a smooth channel wall can be obtained without the need for a post-machining annealing operation by performing the scribing process with the CO₂ laser beam in an unfocused condition. The practicality of the proposed approach is demonstrated by fabricating two microfluidic chips, namely a cytometer, and an integrating microfluidic chip for methanol detection, respectively. The results confirm that the proposed unfocused ablation technique represents a viable solution for the rapid and economic fabrication of a wide variety of PMMA-based microfluidic chips.     

Engineering finger-operated peristaltic pumps

Peristaltic pumping has been widely adopted in microfluidic systems over the past decade. Most applications lie, however, in the regime where fluids or biospecimens are continuously pumped through the entire fluidic system, leaving the initial filling stage of fluid into an air-filled channel underexplored. We propose a compact, quasi-1D, lumped element model for describing the initial filling process of liquid into microfluidic channels driven by peristaltic pumps with discrete diaphragm valves. In addition, we experimentally demonstrated that the liquid penetration length into the fluid channel could be decently controlled (~ 0.3 mm/cycle) using human fingers as the source of actuation pressures. Moreover, we show from our experiments the possibility of controlling the profiles liquid penetration lengths to be other than Lucas–Washburn’s \( l\,\sim\,t^{{\frac{1}{2}}} \) dependence: Linear liquid penetration length profiles can be achieved via liquid channels with exponentially decaying cross-sectional areas. In the above ways, our model has successfully demonstrated that finger-operated peristaltic pumps can serve as an upgraded alternative for capillary-driven microfluidics, offering extra runtime adjustment capability in understanding and engineering of microfluidics in the initial filling stages.     

Sequential enzymatic quantification of two sugars in a single microchannel

This paper presents the implementation of a multiple analyte enzyme assay, based on the sequential injection of the different enzyme solutions, in an electrokinetic driven microfluidic chip. The assay methodology for the simultaneous quantification of d-glucose and d-fructose was reported in previous publications but here the real integration of both enzyme assays was achieved. When assays were executed separately, good reproducibility was observed with average CV values of 5.2% and 4.5% for the d-glucose and d-fructose assay, respectively. Next, the assays for the quantification of d-glucose and d-fructose were integrated simultaneously on chip, where each assay was executed consecutively in the same microreactor by applying a specific sequence of potentials at the reservoirs. This article proves the integration of a sequential based quantification approach in continuous microfluidic chips with electrokinetic actuation.     

A simple and cost-effective method for fabrication of integrated electronic-microfluidic devices using a laser-patterned PDMS layer

We report a simple and cost-effective method for fabricating integrated electronic-microfluidic devices with multilayer configurations. A CO₂ laser plotter was employed to directly write patterns on a transferred polydimethylsiloxane (PDMS) layer, which served as both a bonding and a working layer. The integration of electronics in microfluidic devices was achieved by an alignment bonding of top and bottom electrode-patterned substrates fabricated with conventional lithography, sputtering and lift-off techniques. Processes of the developed fabrication method were illustrated. Major issues associated with this method as PDMS surface treatment and characterization, thickness-control of the transferred PDMS layer, and laser parameters optimization were discussed, along with the examination and testing of bonding with two representative materials (glass and silicon). The capability of this method was further demonstrated by fabricating a microfluidic chip with sputter-coated electrodes on the top and bottom substrates. The device functioning as a microparticle focusing and trapping chip was experimentally verified. It is confirmed that the proposed method has many advantages, including simple and fast fabrication process, low cost, easy integration of electronics, strong bonding strength, chemical and biological compatibility, etc.     

A hybrid approach for fabrication of polymeric BIOMEMS devices

Polymeric microfluidic chips are an enabling component for cost-effective, point of care analytical devices for pharmaceutical, agriculture, health, biological and medical applications. The microfluidic structures can be completed with active elements like pumps and valves as well as sensor components for more complex so called total analysis systems. Often, systems are designed as reader and disposable cartridge where the fluidic structures are simple devices that will be inserted into the reader, which executes the analytical protocol and displays the information in digital form, and disposed after completion of the analysis. In this paper, a hybrid fabrication approach was employed to build a polymeric microfluidic device, so-called sweatstick, suitable for collecting small, precise amounts (600 μl) of human sweat, which were further analyzed for the amount of calcium ions indicating bone mass loss. The device was assembled from different parts fabricated by ultra deep X-ray lithography, precision micro-milling, and molding. Surface treatment of liquid exposed surfaces by oxygen plasma ensures hydrophilic behavior and proper capillary action. Preliminary testing of the device was performed by collecting defined amounts of sweat simulant and determining the calcium ion content using a fluorescent technique. The results for low calcium ion concentration typical for human sweat were excellent and repeatable with variation less than 5% demonstrating the ability to perform indirect bone loss measurements.