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At least 199 records · Page 11Linked to original sources

PubMed2007

Quantitative characterization of magnetic separators: comparison of systems with and without integrated microfluidic mixers.

We present two new types of microfluidic passive magnetic bead separator systems as well as methods for performing quantitative characterizations of them. Both systems consist of a microfluidic channel with long rectangular magnetic elements of permalloy that are placed by the sides of the channel and magnetized by an external magnetic field. In one of the systems, a staggered herringbone microfluidic mixer is integrated in the channel. The characterization of the systems includes magnetic measurements of the capture-and-release efficiencies, estimates of distributions of captured beads in a channel from micrographs, and simulations and analytical models of bead trajectories, capture efficiencies, and capture distributions. We show that the efficiencies of both systems compare favorably to those in the literature. For the studied geometries, the mixer is demonstrated to increase the bead capture-and-release efficiency for a fixed flow rate by up to a factor of two. Moreover, high capture efficiencies can be achieved in the system with mixer at up to ten times higher flow rates than in the system without mixer.

Computer Simulation↗

PubMed2004

Affinity depletion of albumin from human cerebrospinal fluid using Cibacron-blue-3G-A-derivatized photopatterned copolymer in a microfluidic device.

In the context of proteomic research, affinity separations for the prefractionation of complex mixtures, such as cell lysates or human tissues, have become increasingly important. Microfluidic devices have shown significant potential to achieve fast analysis and low sample consumption. Here, we demonstrate the use of a microfluidic device to achieve affinity capture of albumin from human cerebrospinal fluid. Traditional photolithography and wet etching techniques were used to fabricate devices from borosilicate glass wafers. Monolithic porous polymer was prepared in a microfluidic channel by photopolymerization of glycidyl methacrylate and trimethylolpropane trimethacrylate. After derivatization with Cibacron-blue-3G-A, the modified polymer was used to achieve affinity capture of lysozyme and human albumin. Both fluorescence detection and matrix-assisted laser desorption ionization time of flight mass spectrometry were used to validate the results.

Albumins↗

PubMed2004

Calorimetric biosensors with integrated microfluidic channels.

A microfluidic device capable of measuring real-time enthalpy changes of biochemical reactions and thermal properties of biological fluids is presented in this paper. The device consists of a freestanding microthermopile integrated with a glass microfluidic reaction chamber. The p-type polysilicon/gold microthermopiles fabricated on a 2 microm thick thermally isolated membrane showed a sensitivity of 0.94 V/W and a thermal time constant of less than 100 ms. Although the device is not restricted to enzymatic reactions, in this paper measurements of the heat of reaction from the catalytic action of glucose oxidase, catalase, and urease on glucose, hydrogen peroxide, and urea, respectively, are reported. Reactions were performed in open air using liquid batch testing and in enclosed fluidic reaction chamber by continuous flow experiments. A sensitivity of 53.5 microV/M for glucose, 26.5 microV/M for hydrogen peroxide and 17 microV/M for urea was obtained. Detection limit for glucose in the continuous flow mode is approximately 2mM (30 pmol). The aim of this work is to demonstrate the potential of the integrated calorimetric microfluidic device for fundamental thermodynamic studies in biochemical reactions. Using arrays of such devices with immobilized enzymes multi-analyte detection can be accomplished and the effects of interferents from competing substrates can be compensated. This paper presents the design, fabrication and initial testing results from such a microthermopile-based thermal biosensor.

Biosensing Techniques↗

PubMed2004

Microfluidic immunosensor systems.

Immunosensing microfluidic devices are reviewed. Devices are commonly fabricated in glass, silicon, and polymers, with polymers seeing greater attention in recent years. Methods have been developed to immobilize antibodies and other molecules and resist non-specific adsorption through surface modification. The most common detection method is fluorescence, followed by electrochemistry. Various microfluidic designs have been reported for immunoassay applications. The observed trends in microfluidic immunoassay applications closely resemble the trends of general immunoassays, where large molecules are detected principally through a sandwich procedure, while competitive assays are used to detect smaller molecules. The following future trends are suggested: more sensitive detection, increased integration and miniaturization, multianalyte analysis, more robust reagents and devices, and increased functionality of surface treatments.

Biosensing Techniques↗

PubMed2005

Constraining the connectivity of neuronal networks cultured on microelectrode arrays with microfluidic techniques: a step towards neuron-based functional chips.

In vitro culture of small neuronal networks with pre-defined topological features is particularly desirable when the electrical activity of such assemblies can be monitored for long periods of time. Indeed, it is hoped that such networks, with pre-determined connectivity, will provide unique insights into the structure/function relationship of biological neural networks and their properties of self-organization. However, the experimental techniques that have been developed so far for that purpose have either failed to provide very long-term pattern definition and retention, or they have not shown potential for integration into more complex microfluidic devices. To address this problem, three-dimensional microfluidic systems in poly(dimethylsiloxane) (PDMS) were fabricated and used in conjunction with both custom-made and commercially available planar microelectrode arrays (pMEAs). Various types of primary neuronal cell cultures were established inside these systems. Extracellular electrical signals were successfully recorded from all types of cells placed inside the patterns, and this bioelectrical activity was present for several weeks. The advantage of this approach is that it can be further integrated with microfluidic devices and pMEAs to yield, for example, complex neuron-based biosensors or chips for pharmacological screening.

Action Potentials↗

PubMed2005

Sample flow switching techniques on microfluidic chips.

This paper presents an experimental investigation into electrokinetically focused flow injection for bio-analytical applications. A novel microfluidic device for microfluidic sample handling is presented. The microfluidic chip is fabricated on glass substrates using conventional photolithographic and chemical etching processes and is bonded using a high-temperature fusion method. The proposed valve-less device is capable not only of directing a single sample flow to a specified output port, but also of driving multiple samples to separate outlet channels or even to a single outlet to facilitate sample mixing. The experimental results confirm that the sample flow can be electrokinetically pre-focused into a narrow stream and guided to the desired outlet port by means of a simple control voltage model. The microchip presented within this paper has considerable potential for use in a variety of applications, including high-throughput chemical analysis, cell fusion, fraction collection, sample mixing, and many other applications within the micro-total-analysis systems field.

Biosensing Techniques↗

PubMed2006

A microfluidic flow-through device for high throughput electrical lysis of bacterial cells based on continuous dc voltage.

Interest in electrical lysis of biological cells on a microfludic platform has increased because it allows for the rapid recovery of intracellular contents without introducing lytic agents. In this study we demonstrated a simple microfluidic flow-through device which lysed Escherichia coli cells under a continuous dc voltage. The E. coli cells had previously been modified to express green fluorescent protein (GFP). In our design, the cell lysis only happened in a defined section of a microfluidic channel due to the local field amplification by geometric modification. The geometric modification also effectively decreased the required voltage for lysis by several folds. We found that local field strength of 1000-1500 V/cm was required for nearly 100% cell death. This threshold field strength was considerably lower than the value reported in the literature, possibly due to the longer duration of the field [Lee, S.W., Tai, Y.C., 1999. Sens. Actuators A: Phys. 73, 74-79]. Cell lysis was detected by both plate count and fluorescence spectroscopy. The cell membrane was completely disintegrated in the lysis section of the microfluidic device, when the field strength was higher than 2000 V/cm. The devices were fabricated using low-cost soft lithography with channel widths considerably larger than the cell size to avoid clogging and ensure stable performance. Our tool will be ideal for high throughput processing of bacterial cells for chemical analysis of intracellular contents such as DNA and proteins. The application of continuous dc voltage greatly simplified the instrumentation compared to devices using electrical pulses for similar purposes. In principle, the same approach can also be applied for lysis of mammalian cells and electroporative transfection.

Cell Culture Techniques↗

PubMed2006

Microfluidic biofuel cells: the influence of electrode diffusion layer on performance.

Microfluidic biofuel cells exploit the lack of convective mixing at low Reynolds number to eliminate the need for a physical membrane to separate fuel from oxidant. This paper demonstrates how the length and spacing of electrodes within a microchannel, and thus thickness of the diffusion layer, affects the performance of a microfluidic biofuel cell. It was found that splitting a single electrode into two (or more) smaller electrodes and separating them by a distance equal to three times their length prevents the continuous increase in thickness of a diffusion layer. This change results in a 25% increase in maximum power density compared to a single electrode device with identical electroactive area. Furthermore, we found that the maximum current density of a microfluidic biofuel cell operated with different electrode configurations (i.e., length of cathode) closely matches that predicted by theory.

Bioelectric Energy Sources↗

PubMed2006

Flexible manipulation of microfluids using optically regulated adsorption/desorption of hydrophobic materials.

To realize highly integrated micro total analysis systems (microTAS), a simply controlled miniaturized valve should be utilized on microfluidic device. In this paper, we describe the application of photo-induced super-hydrophilicity of titanium dioxide (TiO2) to microfluidic manipulation. In addition, we found a new phenomenon for reversibly converting the surface wettability using a polydimethylsiloxane (PDMS) matrix and the photocatalytic properties of TiO2. While PDMS polymer was irradiated with UV, it was confirmed that hydrophobic material was released from the polymer to air. Several prepolymers were identified as the hydrophobic material with a gas chromatograph and mass spectrometer (GC/MS). Here, we successfully demonstrated the flexible manipulation of microfluid in a branched microchannel using the reversible wettability as micro opto-switching valve (MOS/V). The simultaneous control of MOS/Vs was also demonstrated on a 256-MOS/V integrated disk. The MOS/V promises to be one of the most effective flow switching valves for advanced applications in highly integrated micro/nano fluidics.

Adsorption↗

PubMed2005

Single-cell analysis by electrochemical detection with a microfluidic device.

A novel electrochemical method with a microfluidic device was developed for analysis of single cells. In this method, cell injection, loading and cell lysis, and electrokinetic transportation and detection of intracellular species were integrated in a microfluidic chip with a double-T injector coupled with an end-channel amperometric detector. A single cell was loaded at the double-T injector on the microfluidic chip by using electric field. Then, the docked cell was lysed by a direct current electric field strength of 220 V/cm. The analyte of interest inside the cell was electrokinetically transported to the detection end of separation channel and was electrochemically detected. External standardization was used to quantify the analyte of interest in individual cells. Ascorbic acid (AA) in single wheat callus cells was chosen as the model compound. AA could be directly detected at a carbon fiber disk bundle electrode. The selectivity of electrochemical detection made the electropherogram simple. The technique described here could, in principle, be applied to a variety of electroactive species within single cells.

Ascorbic Acid↗

PubMed2006

Proteins modification of poly(dimethylsiloxane) microfluidic channels for the enhanced microchip electrophoresis.

This report described proteins modification of poly(dimethylsiloxane) (PDMS) microfluidic chip based on layer-by-layer (LBL) assembly technique for enhancing separation efficiency. Two kinds of protein-coated films were prepared. One was obtained by successively immobilizing the cationic polyelectrolyte (chitosan, Chit), gold nanoparticles (GNPs), and protein (albumin, Albu) to the PDMS microfluidic channels surface. The other was achieved by sequentially coating lysozyme (Lys) and Albu. Neurotransmitters (dopamine, DA; epinephrine, EP) and environmental pollutants (p-phenylenediamine, p-PDA; 4-aminophenol, 4-AP; hydroquinone, HQ) as two groups of separation models were studied to evaluate the effect of the functional PDMS microfluidic chips. The results clearly showed these analytes were efficiently separated within 140 s in a 3.7 cm long separation channel and successfully detected with in-channel amperometric detection mode. Experimental parameters in two protocols were optimized in detail. The detection limits of DA, EP, p-PDA, 4-AP, and HQ were 2.0, 4.7, 8.1, 12.3, and 14.8 microM (S/N=3) on the Chit-GNPs-Albu coated PDMS/PDMS microchip, and 1.2, 2.7, 7.2, 9.8, and 12.2 microM (S/N=3) on the Lys-Albu coated one, respectively. In addition, through modification, the more homogenous channel surface displayed higher reproducibility and better stability.

Buffers↗

PubMed2006

Continuous separation of particles using a microfluidic device equipped with flow rate control valves.

We propose herein an improved microfluidic system for continuous and precise particle separation. We have previously proposed a method for particle separation called "pinched flow fractionation." Using the previously reported method, particles can be continuously separated according to differences in their diameters, simply by introducing liquid flows with and without particles into a specific microchannel structure. In this study, we incorporated PDMS membrane microvalves for flow rate control into the microfluidic device to improve the separation accuracy. By adjusting the flow rates distributed to each outlet, target particles could be precisely collected from the desired outlet. We succeeded in separating micron and submicron-size polymer particles. This method can be used widely for continuous and precise separation of various kinds of particles, and can function as an important part of microfluidic systems.

Microfluidic Analytical Techniques↗

PubMed2006

Interfacing capillary gel microfluidic chips with infrared laser desorption mass spectrometry.

We report on the fabrication and performance of a gel microfluidic chip interfaced to laser desorption/ionization (LDI) mass spectrometry with a time-of-flight mass analyzer. The chip was fabricated from poly(methylmethacrylate) with a poly(dimethyl siloxane) cover. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis was performed in the channel of the microfluidic chip. After electrophoresis, the cover was removed and either the PDMS chip or the PMMA cover was mounted in a modified MALDI ion source for analysis. Ions were formed by irradiating the channel with 2.95 microm radiation from a pulsed optical parametric oscillator (OPO), which is coincident with IR absorption by N-H and O-H stretch of the gel components. No matrix was added. The microfluidic chip design allowed a decrease in the volume of material required for analysis over conventional gel slabs, thus enabling improvement in the detection limit to a pmol level, a three orders of magnitude improvement over previous studies in which desorption was achieved from an excised section of a conventional gel.

Comet Assay↗

PubMed2003

Microfluidics in structural biology: smaller, faster em leader better.

Microfluidic technologies promise unprecedented savings in cost and time through the integration of complex chemical and biological assays on a microfabricated chip. Recent advances are making elements of this vision a reality, facilitating the first large-scale integration of microfluidic plumbing with biological assays. The power of miniaturization lies not only in achieving an economy of scale, but also in exploiting the unusual physics of fluid flow and mass transport on small length scales to realize precise and efficient assays that are not accessible with macroscopic tools. Diverse applications ranging from time-resolved studies of protein folding to highly efficient protein crystal growth suggest that microfluidics may become an indispensable tool in biology.

Biological Assay↗

PubMed2005

Microfluidic sonicator for real-time disruption of eukaryotic cells and bacterial spores for DNA analysis.

Biologic agent screening is a three-step process: lysis of host cell membranes or walls to release their DNA, polymerase chain reaction to amplify the genetic material and screening for distinguishing genetic signatures. Macrofluidic devices commonly use sonication as a lysis method. Here, we present a piezoelectric microfluidic minisonicator and test its performance. Eukaryotic human leukemia HL-60 cells and Bacillus subtilis bacterial spores were lysed as they passed through a microfluidic channel at 50 microL/min and 5 microL/min, respectively, in the absence of any chemical denaturants, enzymes or microparticles. We used fluorescence-activated cell sorting and hematocytometry to measure 80% lysis of HL-60 cells after 3 s of sonication. Real-time polymerase chain reaction indicated 50% lysis of B. subtilis spores with 30 s of sonication. Advantages of the minisonicator over macrofluidic implementations include a small sample volume (2.5 microL), reduced energy consumption and compatibility with other microfluidic blocks. These features make this device an attractive option for "lab-on-a-chip" and portable applications.

Bacillus subtilis↗

PubMed2007

Proliferation and viability of adherent cells manipulated by standing-wave ultrasound in a microfluidic chip.

Ultrasonic-standing-wave (USW) technology has potential to become a standard method for gentle and contactless cell handling in microfluidic chips. We investigate the viability of adherent cells exposed to USWs by studying the proliferation rate of recultured cells following ultrasonic trapping and aggregation of low cell numbers in a microfluidic chip. The cells form 2-D aggregates inside the chip and the aggregates are held against a continuous flow of cell culture medium perpendicular to the propagation direction of the standing wave. No deviations in the doubling time from expected values (24 to 48 h) were observed for COS-7 cells held in the trap at acoustic pressure amplitudes up to 0.85 MPa and for times ranging between 30 and 75 min. Thus, the results demonstrate the potential of ultrasonic standing waves as a tool for gentle manipulation of low cell numbers in microfluidic systems.

Animals↗

PubMed2003

Electric field-decoupled electroosmotic pump for microfluidic devices.

An electric field-free electroosmotic pump has been constructed and its pumping rate has been measured under various experimental conditions. The key component of the pump is an ion-exchange membrane grounding joint that serves two major functions: (i) to maintain fluid continuity between pump channels and microfluidic conduit and (ii) to ground the solution in the microfluidic channel at the joint through an external electrode, and hence to decouple the electric field applied to the pump channels from the rest of the microfluidic system. A theoretical model has been developed to calculate the pumping rates and its validity has been demonstrated.

Electricity↗

PubMed2003

Single nucleotide polymorphism analysis by allele-specific primer extension with real-time bioluminescence detection in a microfluidic device.

A microfluidic approach for rapid bioluminescent real-time detection of single nucleotide polymorphism (SNP) is presented. The method is based on single-step primer extension using pyrosequencing chemistry to monitor nucleotide incorporations in real-time. The method takes advantage of the fact that the reaction kinetics differ between matched and mismatched primer-template configurations. We show here that monitoring the initial reaction in real time accurately scores SNPs by comparing the initial reaction kinetics between matched and mismatched configurations. Thus, no additional treatment is required to improve the sequence specificity of the extension, which has been the case for many allele-specific extension assays. The microfluidic approach was evaluated using four SNPs. Three of the SNPs included primer-template configurations that have been previously reported to be difficult to resolve by allele-specific primer extension. All SNPs investigated were successfully scored. Using the microfluidic device, the volume for the bioluminescent assay was reduced dramatically, thus offering a cost-effective and fast SNP analysis method.

Alleles↗