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At least 19 recordsLinked to original sources
PubMed2006

Integration of microcolumns and microfluidic fractionators on multitasking centrifugal microfluidic platforms for the analysis of biomolecules.

This work demonstrates the development of microfluidic compact discs (CDs) for protein purification and fractionation integrating a series of microfluidic features, such as microreservoirs, microchannels, and microfluidic fractionators. The CDs were fabricated with polydimethylsiloxane (PDMS), and each device contained multiple identical microfluidic patterns. Each pattern employed a microfluidic fractionation feature with operation that was based on the redirection of fluid into an isolation chamber as a result of an overflow. This feature offers the advantage of automated operation without the need for any external manipulation, which is independent of the size and the charge of the fractionated molecules. The performance of the microfluidic fractionator was evaluated by its integration into a protein purification microfluidic architecture. The microfluidic architecture employed a microchamber that accommodated a monolithic microcolumn, the fractionator, and an isolation chamber, which was also utilized for the optical detection of the purified protein. The monolithic microcolumn was polymerized "in situ" on the CD from a monolith precursor solution by microwave-initiated polymerization. This technique enabled the fast, efficient, and simultaneous polymerization of monoliths on disposable CD microfluidic platforms. The design of the CD employed allows the integration of various processes on a single microfluidic device, including protein purification, fractionation, isolation, and detection.

Microfluidic Analytical Techniques↗
PubMed2000

Microfluidic tectonics: a comprehensive construction platform for microfluidic systems.

A microfluidic platform for the construction of microscale components and autonomous systems is presented. The platform combines liquid-phase photopolymerization, lithography, and laminar flow to allow the creation of complex and autonomous microfluidic systems. The fabrication of channels, actuators, valves, sensors, and systems is demonstrated. Construction times can be as short as 10 min, providing ultrarapid prototyping of microfluidic systems.

Journal Article↗
PubMed2006

Patterning microbeads inside poly(dimethylsiloxane) microfluidic channels and its application for immobilized microfluidic enzyme reactors.

We propose a convenient and reliable approach for immobilizing microbeads on poly(dimethylsiloxane) (PDMS) microchips. It is built upon a simple fabrication procedure of PDMS chip through directly printing the master with an office laser printer which was described in our previous work (J. Chromatogr. A 2005, 1089, 270-275). On the printed toners used as the positive relief of the master, microbeads were immobilized by a thermal treatment and then transferred to the surface of the microchip by direct molding of the prepolymer on the master. With this approach, the region-selective immobilization of microbeads and the fabrication of PDMS microchips can be accomplished at the same time. Then, using these microbeads as supports, further modification with enzyme was achieved. Surface characteristics of the microbeads-modified PDMS microchannels were investigated with scanning electron microscope, atomic force microscope, and inverse fluorescence microscope. The electrokinetic properties of the native PDMS and the modified PDMS chips were also compared. Based on this approach, an immobilized glucose oxidase (GOD) reactor was constructed and the reaction using glucose as substrate was studied. All these experiments aim to show that the proposed approach may have a good potential in the study of biochemistry and other related areas.

Catalysis↗
PubMed2004

Effect of pressure and fat content on particle sizes in microfluidized milk.

Average diameters and particle size distributions in fluid milks with different fat contents and subjected to various homogenization pressures with a "microfluidizer" were evaluated. Skim, 2%, and whole milks were microfluidized at 50, 100, 150, and 200 MPa. Cream containing 41% milk fat was microfluidized at 50, 100, and 150 MPa. Particle sizes were determined by laser light scattering. As microfluidization pressure was increased from 50 to 100 MPa, particle sizes in skim, 2%, and whole milks decreased. Microfluidization at pressures greater than 100 MPa had little additional effect on reducing the particle sizes in skim and 2% milks compared with microfluidization at 100 MPa, but the particle sizes in whole milk increased as the microfluidization pressure was increased from 100 to 200 MPa due to formation of homogenization clusters. The particle sizes in cream increased as the microfluidization pressure was increased from 50 to 150 MPa. When the microfluidization pressure was held constant, the particle sizes increased as the milk fat concentration was increased. The coefficients of variations of the volume-weighted particle size distributions for cream were higher than for skim, 2%, and whole milks. Larger "big" particles and smaller "small" particles were formed in whole milk after microfluidization at 200 MPa than at 100 MPa. Although microfluidization can be used to produce small particles in skim, 2%, and whole milks, a higher than optimum pressure (above 100 MPa) applied to whole milk will not lead to the minimum d(43) (volume-weighted average diameter) due to formation of clusters.

Animals↗
PubMedDate not supplied

Microfluidic PDMS (polydimethylsiloxane) bioreactor for large-scale culture of hepatocytes.

Microfluidics could provide suitable environments for cell culture because of the larger surface-to-volume ratio and fluidic behavior similar to the environments in vivo. Such microfluidic environments are now used to investigate cell-to-cell interactions and behaviors in vitro, emulating situations observed in vivo, for example, microscale blood vessels modeled by microfluidic channels. These emulated situations cannot be realized by conventional technologies. In our previous works, microfluidic channels composed of two PDMS (poly(dimethylsiloxane)) layers were successfully used for Hep G2 cell culture. To achieve physiologically meaningful functions in vitro, a culture with a larger number of cells and higher density must be performed. This will require bioreactors with larger surface areas for cell attachment and sufficient amounts of oxygen and nutrition supply. For those purposes, we fabricated a bioreactor by stacking 10 PDMS layers together, i.e., four cell culture chambers, and a chamber dedicated to the oxygen supply inserted in the middle of the 10-stacked layers. The oxygen supply chamber is separated from the microfluidic channels for the culture medium perfusion by thin 300-microm PDMS walls. The high gas permeability of PDMS allows oxygen supply to the microfluidic channels through the thin walls. On the basis of the measurement of glucose consumption and albumin production, it is shown that cellular activity exhibits a gradual increase and saturation throughout the culture. We clearly observed that in the case of the microfluidic bioreactor for large-scale cultures, the oxygen chamber is indispensable to achieve longer and healthy cultures. In the present bioreactor, the cell density was found to be about 3-4 x 10(7) cells/cm(3), which is in the same order of magnitude as the conventional macroscale bioreactors. Consequently, by stacking single culture chambers and oxygen chambers in between, we could have a scalable method to realize the microfluidic bioreactor for large-scale cultures.

Albumins↗
PubMed2001

Field-effect flow control in a polydimethylsiloxane-based microfluidic system.

The application of the field-effect for direct control of electroosmosis in a polydimethylsiloxane (PDMS)-based microfluidic system, constructed on a silicon wafer with a 2.0 microm electrically insulating layer of silicon dioxide, is demonstrated. This microfluidic system consists of a 2.0 cm open microchannel fabricated on a PDMS slab, which can reversibly adhere to the silicon wafer to form a hybrid microfluidic device. Aside from mechanically serving as a robust bottom substrate to seal the channel and support the microfluidic system, the silicon wafer is exploited to achieve field-effect flow control by grounding the semiconductive silicon medium. When an electric field is applied through the channel, a radial electric potential gradient is created across the silicon dioxide layer that allows for direct control of the zeta potential and the resulting electroosmotic flow (EOF). By configuring this microfluidic system with two power supplies at both ends of the microchannel, the applied electric potentials can be varied for manipulating the polarity and the magnitude of the radial electric potential gradient across the silicon dioxide layer. At the same time, the longitudinal potential gradient through the microchannel, which is used to induce EOF, is held constant. The results of EOF control in this hybrid microfluidic system are presented for phosphate buffer at pH 3 and pH 5. It is also demonstrated that EOF control can be performed at higher solution pH of 6 and 7.4 by modifying the silicon wafer surface with cetyltrimethylammonium bromide (CTAB) prior to assembly of the hybrid microfluidic system. Results of EOF control from this study are compared with those reported in the literature involving the use of other microfluidic devices under comparable solution conditions.

Cetrimonium↗
PubMedDate not supplied

Experimental modulation of the plasmalemmal microfluidity. Studies on endothelial and aortic smooth muscle cells.

The microfluidity of cell membranes has been modified experimentally in endothelial cells and smooth muscle cells of bovine or monkey aorta cultured in vitro. Microfluidity was estimated by fluorescence depolarization measurements of diphenyl-hexatriene (DPH)-labelled cells. In both types of cells investigated, the arachidonic acid at concentration of 90 microM induced an increase in the microfluidity by 26-53% whereas the cholesterol at the same concentration produced a decrease in the microfluidity by 16-25%. The oleic acid in the range of 30 to 90 microM increased the monkey smooth muscle cell membranes microfluidity by 21-33% but did not change the microfluidity of endothelial and bovine aortic smooth muscle cells. The stearic acid did not influence the microfluidity of either type of cells under investigation. Cortisol at 90 microM changed the microfluidity of the bovine aortic endothelial cells plasmalemma depending on the incubation time. Possible factors of error in the physical measurements due to the extracellular localization of DPH have been identified.

Animals↗
PubMed2005

Microfluidic device for rapid (<15 min) automated microarray hybridization.

BACKGROUND: Current hybridization protocols on microarrays are slow and need skilled personnel. Microfluidics is an emerging science that enables the processing of minute volumes of liquids to perform chemical, biochemical, or enzymatic analyzes. The merging of microfluidics and microarray technologies constitutes an elegant solution that will automate and speed up microarray hybridization. METHODS: We developed a microfluidic flow cell consisting of a network of chambers and channels molded into a polydimethylsiloxane substrate. The substrate was aligned and reversibly bound to the microarray printed on a standard glass slide to form a functional microfluidic unit. The microfluidic units were placed on an engraved, disc-shaped support fixed on a rotational device. Centrifugal forces drove the sample and buffers directly onto the microarray surface. RESULTS: This microfluidic system increased the hybridization signal by approximately 10fold compared with a passive system that made use of 10 times more sample. By means of a 15-min automated hybridization process, performed at room temperature, we demonstrated the discrimination of 4 clinically relevant Staphylococcus species that differ by as little as a single-nucleotide polymorphism. This process included hybridization, washing, rinsing, and drying steps and did not require any purification of target nucleic acids. This platform was sensitive enough to detect 10 PCR-amplified bacterial genomes. CONCLUSION: This removable microfluidic system for performing microarray hybridization on glass slides is promising for molecular diagnostics and gene profiling.

DNA↗
PubMed2004

Single-cell analysis of yeast, mammalian cells, and fungal spores with a microfluidic pressure-driven chip-based system.

BACKGROUND: Cytomics aims at understanding the function of cellular systems by analysis of single cells. Recently, there has been a growing interest in single cell measurements being performed in microfluidic systems. These systems promise to integrate staining, measurement, and analysis in a single system. One important aspect is the limitation of allowable cell sizes due to microfluidic channel dimensions. Here we want to demonstrate the broad applicability of microfluidic chip technology for the analysis of many different cell types. METHODS: We have developed a microfluidic chip and measurement system that allows flow cytometric analysis of fluorescently stained cells from different organisms. In this setup, the cells are moved by pressure-driven flow inside a network of microfluidic channels and are analyzed individually by fluorescence detection. RESULTS: We have successfully applied the system to develop a methodology to detect viable and dead cells in yeast cell populations. Also, we have measured short interfering RNA (siRNA) mediated silencing of protein expression in mammalian cells. In addition, we have characterized the infection state of Magnaportae grisea fungal spores. CONCLUSIONS: Results obtained with the microfluidic system demonstrate a broad applicability of microfluidic flow cytometry to measurement of various cell types.

Cell Line↗
PubMed2006

A label-free protein microfluidic array for parallel immunoassays.

A label-free protein microfluidic array for immunoassays based on the combination of imaging ellipsometry and an integrated microfluidic system is presented. Proteins can be patterned homogeneously on substrate in array format by the microfluidic system simultaneously. After preparation, the protein array can be packed in the microfluidic system which is full of buffer so that proteins are not exposed to denaturing conditions. With simple microfluidic channel junction, the protein microfluidic array can be used in serial or parallel format to analyze single or multiple samples simultaneously. Imaging ellipsometry is used for the protein array reading with a label-free format. The biological and medical applications of the label-free protein microfluidic array are demonstrated by screening for antibody-antigen interactions, measuring the concentration of the protein solution and detecting five markers of hepatitis B.

Animals↗
PubMed2005

Evaluation of microfluidic biosensor development using microscopic analysis of molecular beacon hybridization kinetics.

Molecular beacons, oligonucleotide probes that fluoresce upon hybridization to a target nucleic acid, can be used in microfluidic devices to detect and quantify nucleic acids in solution as well as inside bacterial cells. Three essential steps towards the development of such devices as integrated microfluidic biosensors using molecular beacons were investigated in the present study. First, experiments using real-time confocal microscopy indicated that diffusion of DNA molecular beacons across a 100-mum diameter microfluidic channel took less than one minute after the flow of reagents was stopped. Second, experiments to evaluate hybridization kinetics of DNA molecular beacons with target nucleic acids in solution showed that DNA molecular beacons can be used to characterize hybridization kinetics in real time in microfluidic channels and that hybridization signals approached their maximum in approximately three minutes. Finally, it was demonstrated that peptide nucleic acid molecular beacons can be used to detect bacterial cells in microfluidic devices. These results suggest that the use of microfluidic devices to detect nucleic acids in solution and in bacterial cells is promising and that further development of an integrated microfluidic biosensor for bacterial detection based on this concept is warranted.

Biosensing Techniques↗
PubMed2003

Using bioinspired thermally triggered liposomes for high-efficiency mixing and reagent delivery in microfluidic devices.

High-efficiency mixing is of fundamental importance for the successful development and application of lab-on-a-chip devices. In this report, we present the use of bioinspired thermally triggered liposomes for the controlled delivery and subsequent rapid mixing of reagents in a microfluidic device. In this technique, reagents are encapsulated inside the aqueous interior of liposomes that are dispersed evenly throughout a microfluidic system. Mixing of the encapsulated reagent and reaction do not occur until the reagent is released by a thermal trigger. This approach takes advantage of the dramatically increased lipid membrane permeability of liposomes near the gel-to-liquid phase transition temperature (T(m)) to deliver reagents at a precise location in the microfluidic device through the modulation of temperature. Implementation of this technique requires the encapsulation of the desired reagent in a liposome whose formulation has an appropriate T(m), as well as accurate spatial control of the temperature in the microfluidic device. As the liposomes are uniformly dispersed through the microfluidic channel, mixing occurs quite rapidly upon the release of the reagent. We demonstrate this technique by using several formulations of thermally triggered liposomes to release the hydrophilic fluorescent dyes at controlled locations in a polycarbonate microfluidic device. Additionally, we demonstrate a DNA labeling reaction using liposomes in a capillary-based microfluidic device. Under the conditions studied here, mixing and reaction are complete in approximately 200 microm of channel length. We believe this approach holds great promise for the performance of rapid high-throughput assays and in particular for biological analytes whose native environment is mimicked by the liposome.

DNA↗
PubMed2005

Sensitive detection using microfluidics technology of single cell PCR products from high and low abundance IgH VDJ templates in multiple myeloma.

Human cancer is inherently heterogeneous, so the ability to monitor individual cancer cells at every clinic visit would be a valuable tool. This work describes the first step towards developing handheld and automated devices for molecular and phenotypic analysis of cancer cells. Here, we show that use of capillary electrophoresis to detect PCR product amplified from either transcripts (high abundance template) or genomic DNA (low abundance template) encoding clonotypic immunoglobulin heavy chain VDJ of plasma cells from patients with multiple myeloma. High abundance IgH VDJ transcripts amplified in conventional systems or by capillary electrophoresis through channels on microfluidic chips or, alternatively, PCR product amplified from individual myeloma plasma cells in a single stage RT-PCR reaction was readily detectable on microfluidic chips. For low abundance templates, a nested PCR strategy was needed to detect PCR product by any method. Using microfluidic chips, PCR products amplified from genomic IgH VDJ DNA were detected in six out of eight plasma cells. Comparison of the ABI3100 and the microfluidic chip indicates that approximately 20 times more sample is injected into the ABI 3100 capillary than for the microfluidics chip. Overall, for high and low abundance template in individual cells, the microfluidic separation/detection system is at least as sensitive as the ABI 3100. In the future, integrated microfluidic platforms that incorporate both PCR cycling and product detection on the same chip are likely to exceed conventional systems in sensitivity and speed of genetic analysis by RT-PCR or PCR.

DNA, Neoplasm↗
PubMed2006

Optical and electrochemical detection techniques for cell-based microfluidic systems.

The ability to fabricate microfluidic systems with complex structures and with compatible dimensions between the microfluidics and biological cells have attracted significant attention in the development of microchips for analyzing the biophysical and biochemical functions of cells. Just as cell-based microfluidics have become a versatile tool for biosensing, diagnostics, drug screening and biological research, detector modules for cell-based microfluidics have also undergone major development over the past decade. This review focuses on detection methods commonly used in cell-based microfluidic systems, and provides a general survey and an in-depth look at recent developments in optical and electrochemical detection methods for microfluidic applications for biological systems, particularly cell analysis. Selected examples are used to illustrate applications of these detection systems and their advantages and weaknesses.

Biosensing Techniques↗
PubMed2004

The potential of autofluorescence for the detection of single living cells for label-free cell sorting in microfluidic systems.

A novel method for studying unlabeled living mammalian cells based on their autofluorescence (AF) signal in a prototype microfluidic device is presented. When combined, cellular AF detection and microfluidic devices have the potential to facilitate high-throughput analysis of different cell populations. To demonstrate this, unlabeled cultured cells in microfluidic devices were excited with a 488 nm excitation light and the AF emission (> 505 nm) was detected using a confocal fluorescence microscope (CFM). For example, a simple microfluidic three-port glass microstructure was used together with conventional electroosmotic flow (EOF) to switch the direction of the fluid flow. As a means to test the potential of AF-based cell sorting in this microfluidic device, granulocytes were successfully differentiated from human red blood cells (RBCs) based on differences in AF. This study demonstrated the use of a simple microfabricated device to perform high-throughput live cell detection and differentiation without the need for cell-specific fluorescent labeling dyes and thereby reducing the sample preparation time. Hence, the combined use of microfluidic devices and cell AF may have many applications in single-cell analysis.

Equipment Design↗
PubMed2005

Integration of microfluidics with a four-channel integrated optical Young interferometer immunosensor.

This report describes an optical sensing hybrid system obtained by bonding a microfluidic system to an integrated optical (IO) four-channel Young interferometer (YI) chip. The microfluidic system implemented into a glass plate consists of four microchannels with cross-sectional dimensions of 200 microm x 15 microm. The microfluidic system is structured in such a way that after bonding to the IO chip, each microchannel addresses one sensing window in the four-channel YI sensor. Experimental tests show that the implementation of the microfluidics reduces the response time of the sensor from 100s, as achieved with a bulky cuvette, to 4s. Monitoring the anti-human serum albumine/human serum albumine (alpha-HSA/HSA) immunoreaction demonstrates the feasibility to use the microfluidic sensing system for immunosensing applications. In this case, a better discrimination between the bulk refractive index change and the layer formation can be made, resulting into higher accuracy and offering the prospect of being able to use the kinetics of the immunoreaction. The microfluidic sensing system shows an average phase resolution of 7 x 10(-5) x 2pi for different pairs of channels, which at the given interaction length of 4 mm corresponds to a refractive index resolution of 6 x 10(-8), being equivalent to a protein mass coverage resolution of 20 fg/mm2.

Antibodies↗