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A novel and simplified procedure for patterning hydrophobic and hydrophilic SAMs for microfluidic devices by using UV photolithography.

This work describes how selective patterning of hydrophobic and hydrophilic areas inside microchannels of microfluidic devices can be achieved by combining well-known chemical protocols and standard photolithography equipment (365 nm). Two techniques have been performed and compared. The first technique is based on the preparation of self-assembled monolayers of photocleavable organosilane and the second one on photoassisted grafting (365 nm) of self-assembled monolayers (SAMs) on a silicon or glass substrate. In the first case, we begin with monolayers carrying an o-nitrobenzyl function (hydrophobic area) that is photochemically cleaved, revealing a carboxylic acid group (hydrophilic area). The problem is that the energy necessary to cleave this monolayer is too high and the reaction time is more than 1 h with 50 mW/cm(2) irradiation flux. To overcome this practical disadvantage, we propose another approach that is based on the thiol-ene reaction with benzophenone as photoinitiator. In this approach, a monolayer of mercaptopropyltrimethoxysilane (MPTS) is prepared first. Subsequently, a hydrocarbon chain is photografted locally onto the thiol layer, forming a hydrophobic surface while the reminding unmodified thiol surface is oxidized into sulfonic acid (hydrophilic area). We demonstrated the feasibility of this approach and synthesized high-quality self-assembled monolayers by UV grafting with an irradiation time of 30 s at 365 nm (50 mW/cm(2)). The modified surfaces have been characterized by contact angle measurements, X-ray photoelectron spectroscopy (XPS), AFM, and multiple internal reflection infrared spectroscopy (MIR-FTIR). The difference in the contact angles on the hydrophilic and hydrophobic surfaces reached a remarkable 77 degrees. We have also demonstrated that this method is compatible with selective surface grafting inside microfluidic channels.

Journal Article↗

Viscosimeter on a microfluidic chip.

In this work, a viscosimeter implemented on a microfluidic chip is presented. The physical principle of this system is to use laminar parallel flows in a microfluidic channel. The fluid to be studied flows side by side with a reference fluid of known viscosity. By using optical microscopy, the shape of the interface between both fluids can be determined. Knowing the flow rates of the two liquids and the geometrical features of the channel, the mean shear rate sustained by the fluid and its viscosity can thus be computed. Accurate and precise measurements of the viscosity as a function of the shear rate can be made using less than 300 microL of fluid. Several complex fluids are tested with viscosities ranging from 10(-)(3) to 70 Pa.s.

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Conductivity detection for monitoring mixing reactions in microfluidic devices.

A conductivity detector was coupled to poly(dimethylsiloxane)-glass capillary electrophoresis microchips to monitor microfluidic flow. Electroosmotic flow was investigated with both conductivity detection (CD) and the current monitoring method. No significant variation was observed between these methods, but CD showed a lower relative standard deviation. Gradient mixing experiments were employed to investigate the relationship between the electrolyte conductivity and the electrolyte concentration. A good linear response of conductivity to concentration was obtained for solutions whose difference in concentrations were less than 27 mM. The new system holds great promise for precision mixing in microfluidic devices using electrically driven flows.

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Rapid fabrication of microfluidic devices in poly(dimethylsiloxane) by photocopying.

A very simple and fast method for the fabrication of poly(dimethylsiloxane) (PDMS) microfluidic devices is introduced. By using a photocopying machine to make a master on transparency instead of using lithographic equipment and photoresist, the fabrication process is greatly simplified and speeded up, requiring less than 1.5 h from design to device. Through SEM characterization, any micro-channel network with a width greater than 50 microm and a depth in the range of 8-14 microm can be made by this method. After sealing to a Pyrex glass plate with micromachined platinum electrodes, a microfluidic device was made and the device was tested in FIA mode with on-chip conductometric detection without using either high voltage or other pumping methods.

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Polyimide-based microfluidic devices.

This paper describes the development of polyimide-based microfluidic devices. A layer transfer and lamination technique is used to fabricate flexible microfluidic channels in various shapes and with a wide range of dimensions. High bond strengths can be achieved by cure cycle adaptation and surface treatment of the polyimide layers prior to bonding. The polyimide microchannels can be combined with metallization layers to fabricate electrodes inside and outside channels. The resulting devices can be used for flexible fluidic and electrical connectors, implantable fluid delivery devices, microelectrodes with embedded fluidic channels, chip-based flow cytometry and for a great variety of other applications in medical, chemical or biological research.

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An organic self-regulating microfluidic system.

In this paper we present an organic feedback scheme that merges microfluidics and responsive materials to address several limitations of current microfluidic systems. By using in situ fabrication and by taking advantage of microscale phenomena (e.g., laminar flow, short diffusion times), we have demonstrated feedback control of the output pH in a completely organic system. The system autonomously regulates an output stream at pH 7 under a range of input flow conditions. A single responsive hydrogel component performs the functionality of traditional feedback system components. Vertically stacked laminar flow is used to improve the time response of the hydrogel actuator. A star shaped orifice is utilized to improve the flow characteristics of the membrane/orifice valve. By changing the chemistry of the hydrogel component, the system can be altered to regulate flow based on hydrogels sensitive to temperature, light, biological/molecular, and others.

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Ultra rapid prototyping of microfluidic systems using liquid phase photopolymerization.

We present a method for the ultra rapid prototyping of microfluidic systems using liquid phase photopolymerization, requiring less than 5 min from design to prototype. Microfluidic device fabrication is demonstrated in a universal plastic or glass cartridge. The method consists of the following steps: introduction of liquid prepolymer into the cartridge, UV exposure through a mask to define the channel geometry, removal of unpolymerized prepolymer, and a final rinse. Rapidly fabricated masters for polydimethylsiloxane micromolding are also demonstrated. The master making process is compared to SU-8 50 photoresist processes. Press-on connectors are developed and demonstrated. All materials used are commercially available and low cost. An extension of these methods (mix and match) is presented that allows for maximal design flexibility and integration with a variety of existing fluidic geometries, components, and processes.

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An acoustic wave sensor incorporated with a microfluidic chip for analyzing muscle cell contraction.

We report the fabrication of a microfluidic chip or lab-on-a-chip integrated with a thickness-shear mode (TSM) acoustic wave sensor for muscle cell analysis. The sensor, essentially an AT-cut quartz crystal, serves as a detector for recording changes in acoustic wave properties occurring in an attached cardiomyocyte (single heart muscle cell) during its contraction and relaxation. Presumably, the changes resulted from alterations in viscoelastic properties (e.g. stiffness) of the cells. The effects of excitation electrode size, the presence of a microfluidic channel plate, and liquid loading on the sensor were first examined. Thereafter, muscle cell contraction analysis upon chemical stimuli were described. The potential of the chip for screening of cardiovascular drugs is discussed.

Animals↗

Integration of nanocapillary arrays into microfluidic devices for use as analyte concentrators.

A nanocapillary array was integrated into a microfluidic device and its ability to concentrate analytes was characterized. Through the application of an electric field across the channel, large molecules were concentrated in front of the nanocapillary array, and a concentrated analyte band was ejected from the channel by reversing the polarity of the electric field. The effects of nanocapillary diameter, analyte charge, analyte concentration, analyte plug length, and analyte relative mobility were investigated. Concentration factors up to 300-fold were measured for fluorescein. By concentrating anionic FITC-labeled peptides, it was demonstrated that the magnitude of the electrophoretic mobility did not have a measurable effect on the concentration factor. Therefore, multiple analytes can be concentrated in front of the same nanocapillary array without adjusting the conditions, provided the analytes have the same net charge. In the presence of an electric field, a charge trapping effect was observed; small anionic molecules can be concentrated in front of nanocapillary array with channel diameters which are orders of magnitude above the molecular weight cut-offs for hydrodynamically driven systems. The concentrating process was found to be very efficient for fluorescein, as no leakage through the nanocapillary array or sorption of fluorescein to the nanocapillary array was observed. Due to their flexibility and efficiency, it is anticipated that nanocapillary arrays will find increased utility in electrokinetically driven microfluidic systems.

Electrophoresis↗

CO2 laser microfabrication of an integrated polymer microfluidic manifold for the determination of phosphorus.

A simple colorimetric technique is implemented in a polymer microfluidic manifold. The simple chemistry aids an uncomplicated microchannel design, which is fabricated by CO(2) laser ablation. Issues such as bonding of multiple layers, alignment of micro-fabricated structures and integration of optical components are addressed. A demonstration of a stopped flow regime in the microfluidic manifold is also presented.

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Fabrication of microfluidic mixers and artificial vasculatures using a high-brightness diode-pumped Nd:YAG laser direct write method.

This paper describes a direct write laser technology, which is fast and flexible, for fabricating multiple-level microfluidic channels. A high brightness diode-pumped Nd-YAG laser with slab geometry was used for its excellent beam quality. Channels with flat walls and staggered herringbone ridges on the floor have been successfully fabricated and their ability to perform passive mixing of liquid is discussed. Also, a multi-width multi-depth microchannel has been fabricated to generate biomimetic vasculatures whose channel diameters change according to Murray's law, which states that the cube of the radius of a parent vessel equals the sum of the cubes of the radii of the daughters. The multi-depth architecture allows for flow patterns to resemble physiological vascular systems with lower overall resistance and more uniform flow velocities throughout the network compared to planar patterning techniques which generate uniformly thin channels. The ability to directly fabricate multiple level structures using relatively straightforward laser technology enhances our ability to rapidly prototype complex lab-on-a-chip systems and to develop physiological microfluidic structures for tissue engineering and investigations in biomedical fluidics problems.

Biomimetics↗

Electrically actuated, pressure-driven microfluidic pumps.

In order to make the lab-on-a-chip concept a reality, it is desirable to have an integrated component capable of pumping fluids through microchannels. We have developed novel, electrically actuated micropumps and have integrated them with microfluidic systems. These devices utilize the build-up of electrolysis gases to achieve pressure-driven pumping, only require small voltages (approximately 10 V), and have approximate dimensions of 5 cm x 3 cm x 2 cm. Furthermore, these micropumps are composed of relatively inexpensive materials, and the reversible sealability of their poly(dimethylsiloxane) body to different microfluidic arrays enables repeated uses of the same pump. Under an applied potential of 10 V, three different micropumps had average flow rates of 8-13 microL min(-1) for water being pumped through five different 2 cm-long, 5500 microm(2) cross-sectional-area channels in poly(methyl methacrylate), in approximate agreement with predicted pump rates. We have also evaluated pump operation at the lower applied potential of 8 V and observed an average flow rate of 6.1 microL min(-1) for a pump-channel system. The current micropump design is capable of sustaining pumping pressures in the range of 300 kPa. The various advantages of these micropumps make them well suited for use in lab-on-a-chip analysis techniques.

Microcomputers↗

Microfluidic flow transducer based on the measurement of electrical admittance.

A new flow transducer for measuring the flow rate of a conducting fluid in a microchannel is reported. In this paper, the measure of flow of such fluid under laminar flow conditions based on the change of electrical admittance is established with the aid of a pair of electrodes parallel to the line of flow in a glass-PDMS microfluidic device. This flow sensor is simple in design and can be integrated to most of the microfluidic platforms. The effect of flow rate of the electrolyte, the frequency of the applied ac voltage, the voltage applied across the detector electrodes, and the conductivity of the electrolyte are varied to optimize for high sensitivity. The optimized values are then used to demonstrate the measurements of very low flow rates (<1 nL s(-1)). This flow sensor can be extended towards the measurement of chemical and biochemical buffers and reagents.

Journal Article↗

A new on-chip ESI nozzle for coupling of MS with microfluidic devices.

This paper presents a new on-chip electrospray ionisation (ESI) nozzle, which can be used as an interface for coupling microfluidic devices with mass spectrometric (MS) detection. The nozzle was micromilled in a polymer foil (polymethylmethacrylate (PMMA) 750 microm thick), normally used as a cover for microfluidic chips. The performance of this device was examined in the ESI-MS analysis of the tetrapeptide MRFA (methionine-argenine-phenylalanine-alanine). The spray quality is basically dependent on the inner diameter of the nozzle, beside the part of the organic modifier in the solution to be sprayed. Three different inner nozzle diameters (30, 50, 100 microm) and two different apex angles were investigated. Stable electrospray conditions can be generated with a relative standard deviation less than 10% of the total ion current, and down to a concentration of 0.01 micromol L(-1). The production of this ESI interface is relatively simple for the purpose of a low-cost batch fabrication of miniaturized analytical instruments.

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Development of a novel microfluidic immunoassay for the detection of Helicobacter pylori infection.

The miniaturization of laboratory processes offers substantial advantages over traditional techniques in terms of cost, speed, and potential for multistage automation. To date, only a few studies have reported successful microfluidics-based immunoassays, most of which rely on fluorescence detection technologies. The goal of this study was to develop a poly(dimethylsiloxane) microfluidics-based immunoassay methodology and a versatile colorimetric quantification scheme for the detection of visual colour changes resulting from immune reactions in microchannels. The novel immunoassay technique was applied towards the detection of Helicobacter pylori infection using 20 human serum samples of known infection status, and results compared with conventional nitrocellulose membrane-based dot-ELISA. The microchannel immunoassay reliably detected H. pylori antigens in quantities on the order of 10 ng, which provides a sensitivity of detection comparable to conventional dot-blot assays. Sensitivity was 100%, specificity was 90%, positive predictive value 91%, and negative predictive value 100%, with an overall accuracy of 95%. The software developed generated results that were consistent with visual observations and by automatically taking into account background intensity changes, the software minimized subjectivity. Volumes of solutions used were 100-fold less compared with conventional immunoassays. Miniaturization of the ELISA using this technique provides a means for the accurate diagnosis of microbial infection while minimizing waste production.

Antigens, Bacterial↗

Development and multiplexed control of latching pneumatic valves using microfluidic logical structures.

Novel latching microfluidic valve structures are developed, characterized, and controlled independently using an on-chip pneumatic demultiplexer. These structures are based on pneumatic monolithic membrane valves and depend upon their normally-closed nature. Latching valves consisting of both three- and four-valve circuits are demonstrated. Vacuum or pressure pulses as short as 120 ms are adequate to hold these latching valves open or closed for several minutes. In addition, an on-chip demultiplexer is demonstrated that requires only n pneumatic inputs to control 2(n-1) independent latching valves. These structures can reduce the size, power consumption, and cost of microfluidic analysis devices by decreasing the number of off-chip controllers. Since these valve assemblies can form the standard logic gates familiar in electronic circuit design, they should be useful in developing complex pneumatic circuits.

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Membranes and microfluidics: a review.

The integration of mass transport control by means of membrane functionality into microfluidic devices has shown substantial growth over the last 10 years. Many different examples of mass transport control have been reported, demonstrating the versatile use of membranes. This review provides an overview of the developments in this area of research. Furthermore, it aims to bridge the fields of microfabrication and membrane science from a membrane point-of-view. First the basic terminology of membrane science will be discussed. Then the integration of membrane characteristics on-chip will be categorized based on the used fabrication method. Subsequently, applications in various fields will be reviewed. Considerations for the use of membranes will be discussed and a checklist with selection criteria will be provided that can serve as a starting point for those researchers interested in applying membrane-technology on-chip. Finally, opportunities for microfluidics based on proven membrane technology will be outlined. A special focus in this review is made on the membrane properties of polydimethylsiloxane (PDMS), since this material is frequently used nowadays in master replication.

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Raman spectroscopic monitoring of droplet polymerization in a microfluidic device.

Microfluidic methodologies are becoming increasingly important for rapid formulation and screening of materials, and development of analytical tools for multiple sample screening is a critical step in achieving a combinatorial 'lab on a chip' approach. This work demonstrates the application of Raman spectroscopy for analysis of monomer composition and degree of conversion of methacrylate-based droplets in a microfluidic device. Droplet formation was conducted by flow focusing on the devices, and a gradient of component composition was created by varying the flow rates of the droplet-phase fluids into the microchannels. Raman data were collected using a fiber optic probe from a stationary array of the droplets/particles on the device, followed by partial least squares (PLS) calibration of the first derivative (1600 cm(-1) to 1550 cm(-1)) allowing successful measurement of monomer composition with a standard error of calibration (SEC) of +/-1.95% by volume. Following photopolymerization, the percentage of double bond conversion of the individual particles was calculated from the depletion of the normalized intensity of the C[double bond, length as m-dash]C stretching vibration at 1605 cm(-1). Raman data allowed accurate measurement of the decrease in double bond conversion as a function of increasing crosslinker concentration. The results from the research demonstrate that Raman spectroscopy is an effective, on-chip analytical tool for screening polymeric materials on the micrometre scale.

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