Showing papers in "Lab on a Chip in 2003"

Rapid droplet mixers for digital microfluidic systems.

Abstract: The mixing of analytes and reagents for a biological or chemical lab-on-a-chip is an important, yet difficult, microfluidic operation. As volumes approach the sub-nanoliter regime, the mixing of liquids is hindered by laminar flow conditions. An electrowetting-based linear-array droplet mixer has previously been reported. However, fixed geometric parameters and the presence of flow reversibility have prevented even faster droplet mixing times. In this paper, we study the effects of varying droplet aspect ratios (height ∶ diameter) on linear-array droplet mixers, and propose mixing strategies applicable for both high and low aspect ratio systems. An optimal aspect ratio for four electrode linear-array mixing was found to be 0.4, with a mixing time of 4.6 seconds. Mixing times were further reduced at this ratio to less than three seconds using a two-dimensional array mixer, which eliminates the effects of flow reversibility. For lower aspect ratio (≤0.2) systems, we present a split-and-merge mixer that takes advantage of the ability to perform droplet splitting at these ratios, resulting in a mixing time of less than two seconds.

Integrating advanced functionality in a microfabricated high-throughput fluorescent-activated cell sorter

Abstract: The integration of complete analyses systems "on chip" is one of the great potentials of microfabricated devices. In this study we present a new pressure-driven microfabricated fluorescent-activated cell sorter chip with advanced functional integration. Using this sorter, fluorescent latex beads are sorted from chicken red blood cells, achieving substantial enrichments at a sample throughput of 12000 cells s(-1). As a part of the sorter chip, we have developed a monolithically integrated single step coaxial flow compound for hydrodynamic focusing of samples in flow cytometry and cell sorting. The structure is simple, and can easily be microfabricated and integrated with other microfluidic components. We have designed an integrated chamber on the chip for holding and culturing of the sorted cells. By integrating this chamber, the risk of losing cells during cell handling processes is eliminated. Furthermore, we have also developed integrated optics for cell detection. Our new design contributes to the ongoing efforts for building a fully integrated micro cell sorting and analysing system.

Electrowetting-based droplet mixers for microfluidic systems

Abstract: Mixing of analytes and reagents is a critical step in realizing a lab-on-a-chip. However, mixing of liquids is very difficult in continuous flow microfluidics due to laminar flow conditions. An alternative mixing strategy is presented based on the discretization of liquids into droplets and further manipulation of those droplets by electrowetting. The interfacial tensions of the droplets are controlled with the application of voltage. The droplets act as virtual mixing chambers, and mixing occurs by transporting the droplet across an electrode array. We also present an improved method for visualization of mixing where the top and side views of mixing are simultaneously observed. Microliters of liquid droplets are mixed in less than five seconds, which is an order of magnitude improvement in reported mixing times of droplets. Flow reversibility hinders the process of mixing during linear droplet motion. This mixing process is not physically confined and can be dynamically reconfigured to any location on the chip to improve the throughput of the lab-on-a-chip.

Enhancement of microfluidic mixing using time pulsing.

Abstract: Many microfluidic applications require the mixing of reagents, but efficient mixing in these laminar (i.e., low Reynolds number) systems is typically difficult. Instead of using complex geometries and/or relatively long channels, we demonstrate the merits of flow rate time dependency through periodic forcing. We illustrate the technique by studying mixing in a simple “T” channel intersection by means of computational fluid dynamics (CFD) as well as physically mixing two aqueous reagents. The “T” geometry selected consists of two inlet channel segments merging at 90 degrees to each other, the outlet segment being an extension of one of the inlet segments. All channel segments are 200 µm wide by 120 µm deep, a practical scale for mass-produced disposable devices. The flow rate and average velocity after the confluence of the two reagents are 48 nl s−1 and 2 mm s−1 respectively, which, for aqueous solutions at room temperature, corresponds to a Reynolds number of 0.3. We use a mass diffusion constant of 10−10 m2 s−1, typical of many BioMEMS applications, and vary the flow rates of the reagents such that the average flow rate remains unchanged but the instantaneous flow rate is sinusoidal (with a DC bias) with respect to time. We analyze the effect of pulsing the flow rate in one inlet only as well as in the two inlets, and demonstrate that the best results occur when both inlets are pulsed out of phase. In this case, the interface is shown to stretch, retain one fold, and sweep through the confluence zone, leading to good mixing within 2 mm downstream of the confluence, i.e. about 1 s of contact. From a practical viewpoint, the case where the inlets are 180 degrees out of phase is of particular interest as the outflow is constant.

Joule heating and heat transfer in poly(dimethylsiloxane) microfluidic systems

Abstract: Joule heating is a significant problem in electrokinetically driven microfluidic chips, particularly polymeric systems where low thermal conductivities amplify the difficulty in rejecting this internally generated heat. In this work, a combined experimental (using a microscale thermometry technique) and numerical (using a 3D "whole-chip" finite element model) approach is used to examine Joule heating and heat transfer at a microchannel intersection in poly(dimethylsiloxane)(PDMS), and hybrid PDMS/Glass microfluidic systems. In general the numerical predictions and the experimental results agree quite well (typically within +/- 3 degree C), both showing dramatic temperature gradients at the intersection. At high potential field strengths a nearly five fold increase in the maximum buffer temperature was observed in the PDMS/PDMS chips over the PDMS/Glass systems. The detailed numerical analysis revealed that the vast majority of steady state heat rejection is through lower substrate of the chip, which was significantly impeded in the former case by the lower thermal conductivity PDMS substrate. The observed higher buffer temperature also lead to a number of significant secondary effects including a near doubling of the volume flow rate. Simple guidelines are proposed for improving polymeric chip design and thereby extend the capabilities of these microfluidic systems.

Reagentless mechanical cell lysis by nanoscale barbs in microchannels for sample preparation

Abstract: A highly effective, reagentless, mechanical cell lysis device integrated in microfluidic channels is reported. Sample preparation, specifically cell lysis, is a critical element in ‘lab-on-chip’ applications. However, traditional methods of cell lysis require purification steps or complicated fabrication steps that a simple mechanical method of lysis may avoid. A simple and effective mechanical cell lysis system is designed, microfabricated, and characterized to quantify the efficiency of cell lysis and biomolecule accessibility. The device functionality is based on a microfluidic filter region with nanostructured barbs created using a modified deep reactive ion etching process. Mechanical lysis is characterized by using a membrane impermeable dye. Three main mechanisms of micro-mechanical lysis are described. Quantitative measurements of accessible protein as compared to a chemically lysed sample are acquired with optical absorption measurements at 280 and 414 nm. At a flow rate of 300 µL min−1 within the filter region total protein and hemoglobin accessibilities of 4.8% and 7.5% are observed respectively as compared to 1.9% and 3.2% for a filter without nanostructured barbs.

Electro-hydrodynamic micro-fluidic mixer

Abstract: Fluid mixing in microchannels is needed for many applications ranging from bio-arrays to micro-reactors, but is typically difficult to achieve. A simple geometry micro-mixer is proposed based on the electro-hydrodynamic (EHD) force present when the fluids to be mixed have different electrical properties and are subjected to an electric field. The electrodes are arranged so that the electric field is perpendicular to the interface between the two fluids, creating a transversal secondary flow. The technique is demonstrated experimentally using the flow of two liquids with identical viscosity and density, but different electrical properties. The volume flow rate and average velocity are 0.26 µl s−1 and 4.2 mm s−1, respectively, corresponding to a Reynolds number Re = 0.0174. The effect of a continuous (DC) electric field and two alternating (AC) – sinusoidal and square – electric fields is explored. At the appropriate parameter values, very good mixing takes place in less than 0.1 s, over a very short distance (within a fraction of the width 250 µm of the electrodes).

PDMS 2D optical lens integrated with microfluidic channels: principle and characterization.

Abstract: In this paper, the fabrication and characterization of PDMS 2D-optical lenses are reported. These lenses are designed in order to improve the performance of fluorescent spectroscopy detection performed on a portable chip using optical fibers. The fabrication process of the PDMS layer is first detailed, and the patterns are then checked with a SEM. By comparing various interfacial structures, it is shown that the beam properties of the light coming out from the fiber can be modified depending on the lens curvature radius. As a consequence, for a constant dye concentration, the use of such lenses can increase the intensity of fluorescent response close to the fiber or far from the fiber, compared to the same design with a flat interface. This excitation improvement corresponding to a stronger response from the dye then consequently leads to around three times higher sensitivity of the on-chip detection method for fluorescent spectroscopy.

Generation of larger numbers of separated microbial populations by cultivation in segmented-flow microdevices

Abstract: The high speed production of fluid segments for the highly parallelized cultivation of monoclonal cell populations was carried out by the use of microchip segmentor modules. Aqueous fluid segments, embedded in a non-miscible carrier liquid, were produced with frequencies up to 30 s−1 and showed a high homogeneity in size. This corresponds with the production of about 2.5 million samples per day. The segment volumes can be adapted between about 4 nl and 100 nl. The typical segment size for cultivation experiments is in the range between 40 nl and 80 nl. Nutrient medium can be applied instead of pure water. It is possible to aliquot a cell suspension in such a way that most of the aqueous fluid segments contain only one cell. In model experiments with four microbial species chip-produced aliquots of 60 nl, each containing one or a few cells, were incubated in Teflon capillary tubes. Rapid growth of the microcultures was observed. Cell densities were found to be as high as in conventional shake flask cultures.

In situ fabricated porous filters for microsystems.

Abstract: Porous filters were fabricated inside a microchannel using emulsion photo-polymerization. The porosity of the filter is dependent on the composition of the pre-polymer mixture. The morphology and flow resistance of four filters with varying water and cross-linker concentrations was studied. The porous filter was used to separate samples, such as whole blood to cell/serum components, based on size. The efficiency of cell/serum separation by the porous filter was tested by performing a glucose-6-phosphate dehydrogenase assay and was found to be comparable to separation by centrifuge. The porous filter thus mimics the functionality of the centrifuge, with the added benefits of no power requirements and ability to handle small sample volumes.

Microfabricated PCR-electrochemical device for simultaneous DNA amplification and detection

Abstract: Microfabricated silicon/glass-based devices with functionalities of simultaneous polymerase chain reaction (PCR) target amplification and sequence-specific electrochemical (EC) detection have been successfully developed. The microchip-based device has a reaction chamber (volume of 8 µl) formed in a silicon substrate sealed by bonding to a glass substrate. Electrode materials such as gold and indium tin oxide (ITO) were patterned on the glass substrate and served as EC detection platforms where DNA probes were immobilized. Platinum temperature sensors and heaters were patterned on top of the silicon substrate for real-time, precise and rapid thermal cycling of the reaction chamber as well as for efficient target amplification by PCR. DNA analyses in the integrated PCR-EC microchip start with the asymmetric PCR amplification to produce single-stranded target amplicons, followed by immediate sequence-specific recognition of the PCR product as they hybridize to the probe-modified electrode. Two electrochemistry-based detection techniques including metal complex intercalators and nanogold particles are employed in the microdevice to achieve a sensitive detection of target DNA analytes. With the integrated PCR-EC microdevice, the detection of trace amounts of target DNA (as few as several hundred copies) is demonstrated. The ability to perform DNA amplification and EC sequence-specific product detection simultaneously in a single reaction chamber is a great leap towards the realization of a truly portable and integrated DNA analysis system.

Design and simulation of the micromixer with chaotic advection in twisted microchannels.

Abstract: Chaotic mixers with twisted microchannels were designed and simulated numerically in the present study. The phenomenon whereby a simple Eulerian velocity field may generate a chaotic response in the distribution of a Lagrangian marker is termed chaotic advection. Dynamic system theory indicates that chaotic particle motion can occur when a velocity field is either two-dimensional and time-dependent, or three-dimensional. In the present study, micromixers with three-dimensional structures of the twisted microchannel were designed in order to induce chaotic mixing. In addition to the basic T-mixer, three types of micromixers with inclined, oblique and wavelike microchannels were investigated. In the design of each twisted microchannel, the angle of the channels' bottoms alternates in each subsection. When the fluids enter the twisted microchannels, the flow sways around the varying structures within the microchannels. The designs of the twisted microchannels provide a third degree of freedom to the flow field in the microchannel. Therefore, chaotic regimes that lead to chaotic mixing may arise. The numerical results indicate that mixing occurs in the main channel and progressively larger mixing lengths are required as the Peclet number increased. The swaying of the flow in the twisted microchannel causes chaotic advection. Among the four micromixer designs, the micromixer with the inclined channel most improved mixing. Furthermore, using the inclined mixer with six subsections yielded optimum performance, decreasing the mixing length by up to 31% from that of the basic T-mixer.

Functional cellular assays with multiparametric silicon sensor chips

Abstract: Multiparametric silicon sensor chips mounted into biocompatible cell culture units have been used for investigations on cellular microphysiological patterns. Potentiometric, amperometric and impedimetric microsensors are combined on a common cell culture surface on the chip with an area of ∼29 mm2. Extracellular acidification rates (with pH-sensitive field effect transistors, ISFETs), cellular oxygen consumption rates (with amperometric electrode structures) and cell morphological alterations (with impedimetric electrode structures, IDES) are monitored on single chips simultaneously for up to several days. The corresponding test device accommodates six of such sensor chips in parallel, provides electronic circuitry and maintains the required cell culture conditions (temperature, fluid perfusion system). Sensor data are transformed into quantitative information about microphysiologic conditions. The outcome of this transformation as well as reliability and sensitivity in detection of drug effects is discussed. This is the first report on multiparametric cell based assays with data obtained solely with integrated sensors on silicon chips. Those assays are required in different fields of application such as pharmaceutical drug screening, tumor chemosensitivity tests and environmental monitoring.

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

Abstract: 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.

Counting and sizing of particles and particle agglomerates in a microfluidic device using laser light scattering: application to a particle-enhanced immunoassay

Abstract: A microfluidic device for counting and sizing particles and particle agglomerates based on laser light scattering is demonstrated. The particles were confined hydrodynamically and passed through a focused laser beam. Scattering at two different angles, 15 degree and 45 degree, was detected. At an acquisition rate of 10 kHz, a throughput of 150 particles s(-1) was achieved. Scattering intensity was found to depend on particle volume for 2 to 9 microm diameter particles. Size discrimination of particles with a diameter ratio of 1: 2 was accomplished. In addition, the scattering signals of particle agglomerates formed in a particle-enhanced immunoassay for C-reactive protein (CRP) were measured. Scattering intensity was found to be dependent on the CRP concentration, 100 ng CRP per mL could be detected. The particle counting method presented is generic and can be employed in a wide variety of assays as well as for cell counting and particle counting.

A polymer-based microfluidic device for immunosensing biochips.

Abstract: This paper describes the design, fabrication, and test of a PDMS/PMMA-laminated microfluidic device for an immunosensing biochip. A poly(dimethyl siloxane)(PDMS) top substrate molded by polymer casting and a poly(methyl methacrylate)(PMMA) bottom substrate fabricated by hot embossing are bonded with pressure and hermetically sealed. Two inlet ports and an air vent are opened through the PDMS top substrate, while gold electrodes for electrochemical biosensing are patterned onto the PMMA bottom substrate. The analyte sample is loaded from the sample inlet port to the detection chamber by capillary force, without any external intervening forces. For this and to control the time duration of sample fluid in each compartment of the device, including the inlet port, diffusion barrier, reaction chamber, flow-delay neck, and detection chamber, the fluid conduit has been designed with various geometries of channel width, depth, and shape. Especially, the fluid path has been designed so that the sample flow naturally stops after filling the detection chamber to allow sufficient time for biochemical reaction and subsequent washing steps. As model immunosensing tests for the microfluidic device, functionalizations of ferritin and biotin to the sensing surfaces on gold electrodes and their biospecific interactions with antiferritin antiserum and streptavidin have been investigated. An electrochemical detection method for immunosensing by biocatalyzed precipitation has been developed and applied for signal registration. With the biochip, the whole immunosensing processes could be completed within 30 min.

Synthesis and analysis of combinatorial libraries performed in an automated micro reactor system

Abstract: This paper presents the synthesis of combinatorial libraries performed on a single-channel glass micro reactor under hydrodynamic flow control. The experiments were carried out in a non-well based micro chip and consisted of the preparation of libraries of pyrazoles by means of a Knorr reaction of 1,3-dicarbonyl compounds with hydrazines. The aim of this work is to investigate the capabilities of an automated micro reactor based system to synthesise sequentially multiple analogue reactions. Small slugs of reactants were introduced automatically by an autosampler in a serpentine-etched glass chip. The mobility of the reagents and products was achieved using hydrodynamic driven flow. Reaction slug dilution and UV slug detection took place at the outlet. A sample of the slug was analysed by using an on-line LC-UV-MS system. The degree of conversion was quantified using the UV signal and comparing with standards of starting materials and final products. After the LC-UV-MS analysis, the automated system proceeds to inject the slugs to carry out the next reaction programmed. The results suggest that the micro reactor system is capable of repeating the process of injection, mixing and reaction in an automated manner as many times as required.

Fabrication of thermoset polyester microfluidic devices and embossing masters using rapid prototyped polydimethylsiloxane molds.

Abstract: Plastics are increasingly being used for the fabrication of Lab-on-a-Chip devices due to the variety of beneficial material properties, affordable cost, and straightforward fabrication methods available from a range of different types of plastics. Rapid prototyping of polydimethylsiloxane (PDMS) devices has become a well-known process for the quick and easy fabrication of microfluidic devices in the research laboratory; however, PDMS is not always an appropriate material for every application. This paper describes the fabrication of thermoset polyester microfluidic devices and masters for hot embossing using replica molding techniques. Rapid prototyped PDMS molds are convienently used for the production of non-PDMS polymeric devices. The recessed features in the cast polyester can be bonded to a second polyester piece to form an enclosed microchannel. Thermoset polyester can withstand moderate amounts of pressure and elevated temperature; therefore, the cast polyester piece also can be used as a master for embossing polymethylmethacrylate (PMMA) microfluidic systems. Examples of enclosed polyester and PMMA microchannels are presented, and we discuss the electroosmotic properties of both types of channels, which are important for analytical applications such as capillary electrophoresis.

Sub-second isoelectric focusing in free flow using a microfluidic device.

Abstract: Using a microfabricated chip with a bed volume of 0.2 µL we demonstrate the validity of the scaling laws for molecular mass transport of isoelectric focusing (IEF) in free flow. Nano- or microlitre sample volumes can be concentrated within 430 ms by a factor of up to 400. These very fast performances make the chip applicable to proteomic analysis and for continuous monitoring of biochemical processes.

Nonlinear decrease of background fluorescence in polymer thin-films – a survey of materials and how they can complicate fluorescence detection in μTAS

Abstract: Polymers and plastics are receiving increased attention as materials for microfluidics and μTAS applications. Given the ubiquity of fluorescence detection techniques in micro-analytical systems, the fluorescence properties of polymers and plastics should not be overlooked. We survey some commonly available polymer thin-films for their fluorescence behaviour under standardized conditions to determine which materials are most suitable for high-sensitivity fluorescence detection lab chips. The initial fluorescence intensities of some of the materials surveyed were significantly higher than glass and fused silica controls, and decreased over the three hour period with complex kinetics. We then discuss how this has confounded fluorescence detection in our analytical context, and possible mechanisms for the decrease.

Microfluidic devices for fluidic circulation and mixing improve hybridization signal intensity on DNA arrays

Abstract: Reactions of biomolecules with surface mounted materials on microscope slides are often limited by slow diffusion kinetics, especially in low volumes where diffusion is the only means of mixing. This is a particular problem for reactions where only small amounts of analyte are available and the required reaction volume limits the analyte concentration. A low volume microfluidic device consisting of two interconnected 9 mm × 37.5 mm reaction chambers was developed to allow mixing and closed loop fluidic circulation over most of the surface of a microscope slide. Fluid samples are moved from one reaction chamber to the other by the rotation of a magnetic stirring bar that is driven by a standard magnetic stirrer. We demonstrate that circulation and mixing of different reagents can be efficiently accomplished by this closed loop device with solutions varying in viscosity from 1 to 16.2 centipoise. We also show by example of a microarray hybridization that the reaction efficiency can be enhanced 2–5 fold through fluid mixing under conditions where diffusion is rate limiting. For comparison, similar results were achieved with a disposable commercial device that covers only half of the reaction area of the closed loop device.

Influence of master fabrication techniques on the characteristics of embossed microfluidic channels

Abstract: We describe protocols for the fabrication of microfluidic devices in plastics using a number of different embossing masters. Masters were fabricated by deep reactive ion etching (DRIE) of silicon (100), wet etching of silicon (100) and (110), and SU-8 processing. Structures embossed into a cyclo-olefin polymer were characterized in terms of the quality of pattern transfer as well as of the surface roughness. High quality pattern transfer was achieved with masters containing structures with angled sidewalls. Pattern distortions occurring during de-embossing were minimized by using masters consisting of SU-8 (which has a thermal expansion coefficient close to that of the substrates). Structures embossed with SU-8 masters also exhibited the lowest surface roughness. However, due to structural deformation, the reusability of the masters prepared for this study extended to only five embossing experiments. Masters fabricated on silicon, on the other hand, were more robust, but were subject to breakage during the de-embossing phase of the experiment. The results of this study will guide researchers in choosing master fabrication methods that will provide profile and surface characteristics of embossed microfluidic channels that are advantageous to their specific application.

Enzymatic degradation of p-chlorophenol in a two-phase flow microchannel system

Abstract: Enzymatic degradation of p-chlorophenol was carried out in a two-phase flow in a microchannel (100 microm width, 25 microm depth) fabricated on a glass plate (70 mm x 38 mm). This is the first report on the enzymatic reaction in a two-phase flow on a microfluidic device. The surface of the microchannel was partially modified with octadecylsilane groups to be hydrophobic, thus allowing clear phase separation at the end-junction of the microchannel. The enzyme (laccase), which is surface active, was solubilized in a succinic aqueous buffer and the substrate (p-chlorophenol) was in isooctane. The degradation of p-chlorophenol occurred mainly at the aqueous-organic interface in the microchannel. We investigated the effects of flow velocity and microchannel shape on the enzymatic degradation of p-chlorophenol. Assuming that diffusion of the substrate (p-chlorophenol) is the rate-limiting step in the enzymatic degradation of p-chlorophenol in the microchannel, we proposed a simple theoretical model for the degradation in the microchannel. The calculated degradation values agreed well with the experimental data.

Chemical and physical processes for integrated temperature control in microfluidic devices

Abstract: Microfluidic devices are a promising new tool for studying and optimizing (bio)chemical reactions and analyses. Many (bio)chemical reactions require accurate temperature control, such as for example thermocycling for PCR. Here, a new integrated temperature control system for microfluidic devices is presented, using chemical and physical processes to locally regulate temperature. In demonstration experiments, the evaporation of acetone was used as an endothermic process to cool a microchannel. Additionally, heating of a microchannel was achieved by dissolution of concentrated sulfuric acid in water as an exothermic process. Localization of the contact area of two flows in a microfluidic channel allows control of the position and the magnitude of the thermal effect.

A microchip-based proteolytic digestion system driven by electroosmotic pumping.

Abstract: Microchip-based proteomic analysis requires proteolytic digestion of proteins in microdevices. Enzyme reactors in microdevices, fabricated in glass, silicon, and PDMS substrates, have recently been demonstrated for model protein digestions. The common approach used for these enzyme reactors is employment of a syringe pump(s) to generate hydrodynamic flow, driving the proteins through the reactors. Here we present a novel approach, using electroosmotic flow (EOF) to electrokinetically pump proteins through a proteolytic system. The existence of EOF in the proteolytic system packed with immobilized trypsin gel beads was proven by imaging the movement of a neutral fluorescent marker. Digestions of proteins were subsequently carried out for 12 min, and the tryptic peptides were analyzed independently using capillary electrophoresis (CE) and MALDI-TOF mass spectrometry (MS). The results from CE analysis of the tryptic peptides from the EOF-driven proteolytic system and a conventional water bath digestion were comparable. MALDI-TOF MS was used to identify the parent protein and the tryptic peptides using MS-Fit database searching. The potential utility of the EOF-driven proteolytic system was demonstrated by direct electro-elution of proteins from an acrylamide gel into the proteolytic system, with elution and tryptic digestion achieved in a single step. The EOF-driven proteolytic system, thus, provides a simple way to integrate protein digestion into an electrophoretic micro total analysis system for protein analysis and characterization.

A functional on-chip pressure generator using solid chemical propellant for disposable lab-on-a-chip

Abstract: This paper presents a functional on-chip pressure generator that utilizes chemical energy from a solid chemical propellant to perform fluidic delivery in applications of plastic-based disposable biochips or lab-on-a-chip systems. In this functional on-chip pressure generator, azobis-isobutyronitrile (AIBN) as the solid chemical propellant is deposited on a microheater using a screen-printing technique, which can heat the AIBN at 70 °C to produce nitrogen gas. The output pressure of nitrogen gas, generated from the solid chemical propellant, is adjustable to a desired pressure by controlling the input power of the heater. Using this chemical energy source, the generated pressure depends on the deposited amount of the solid chemical propellant and the temperature of the microheater. Experimental measurements show that this functional on-chip pressure generator can achieve around 3 000 Pa pressure when 189 mJ of energy is applied to heat the 100 µg of AIBN. This pressure can drive 50 nl of water through a microfluidic channel of 70 mm and cross-sectional area of 100 µm × 50 µm. Due to its compact size, ease of fabrication and integration, high reliability (no moving parts), biologically inert gas output along with functionality of gas generation, this pressure generator will be an excellent pressure source for handling the fluids of disposable lab-on-a-chip, biochemical analysis systems or drug delivery systems.

Poly(dimethylsiloxane) electrospray devices fabricated with diamond-like carbon–poly(dimethylsiloxane) coated SU-8 masters

Abstract: This study presents coupling of a poly(dimethylsiloxane) (PDMS) micro-chip with electrospray ionization-mass spectrometry (ESI-MS). Stable electrospray is generated directly from a PDMS micro-channel without pressure assistance. Hydrophobic PDMS aids the formation of a small Taylor cone in the ESI process and facilitates straightforward and low-cost batch production of the ESI-MS chips. PDMS chips were replicated with masters fabricated from SU-8 negative photoresist. A novel coating, an amorphous diamond-like carbon–poly(dimethylsiloxane) hybrid, deposited on the masters by the filtered pulsed plasma arc discharge technique, improved significantly the lifetime of the masters in PDMS replications. PDMS chip fabrication conditions were observed to affect the amount of background peaks in the MS spectra. With an optimized fabrication process (PDMS curing agent/silicone elastomer base ratio of 1/8 (w/w), curing at 70 °C for 48 h) low background spectra were recorded for the analytes. The performance of PDMS devices was examined in the ESI-MS analysis of some pharmaceutical compounds and amino acids.