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At least 325 records · Page 18Linked to original sources

PubMed2000

Electrokinetically controlled microfluidic analysis systems.

Electrokinetic forces are emerging as a powerful means to drive microfluidic systems with flow channel cross-sectional dimensions in the tens of micrometers and flow rates in the nanoliter per second range. These systems provide many advantages such as improved analysis speed, improved reproducibility, greatly reduced reagent consumption, and the ability to perform multiple operations in an integrated fashion. Planar microfabrication methods are used to make these analysis chips in materials such as glass or polymers. Many applications of this technology have been demonstrated, such as DNA separations, enzyme assays, immunoassays, and PCR amplification integrated with microfluidic assays. Further development of this technology is expected to yield higher levels of functionality of sample throughput on a single microfluidic analysis chip.

Chemistry Techniques, Analytical↗

PubMed2004

Integrated system for rapid proteomics analyses using microfluidic devices coupled to nanoelectrospray mass spectrometry.

This chapter presents an integrated and modular microsystem providing rapid analyses of low femtomole of in-gel digests for proteomics applications. Enhancement of sample throughput is facilitated using an autosampler, a microfabricated device comprising a large (2.4-microL total volume) separation channel together with a low-dead-volume interface to nES mass spectrometry. Sample preconcentration is achieved by packing C18 reverse phase or immobilized metal affinity chromatography (IMAC) beads into the large channel of this microfluidic device to adsorb peptides or enrich the sample in phosphopeptides prior to capillary electrophoresis separation and MS detection. This integrated microfluidic systems enables a sample throughput of up to 12 samples/h with a detection limit of approx 5 nM (25 fmol inj.). Replicate injections of peptide standards indicated that reproducibility of migration time was typically 1.2-1.8%, whereas relative standard deviation (RSD) values of 9.2-11.8% were obtained on peak heights. The application of this device is demonstrated for 2D gel spots obtained from protein extracts of human astrocyte cells and for excised bands of membrane proteins from Neisseria meningitidis. A stepped acetonitrile gradient can be incorporated with the present microfluidic system to enhance selectivity during sample analysis.

Astrocytes↗

PubMed2003

Comparison of oil-in-water emulsions manufactured by microfluidization and homogenization.

The purpose of this study was to compare drug-free model submicron oil-in-water (o/w) emulsions manufactured by high-speed homogenization and microfluidization. The study was aimed at evaluating the influence of these two manufacturing processes on the stability of the emulsions with respect to emulsifier concentration. Stability was defined in terms of dispersed droplet diameter growth over time. The study was also directed towards identifying the minimum emulsifier concentrations required by either processing method within the same model o/w systems to produce emulsions viable throughout the study period of three months. The Microfluidizer 110L was found to be more effective than the homogenizer in producing stable o/w submicron emulsions using triglycerides of caprylic/capric acid as the oil phase and combinations of emulsifiers (polyoxyethylene sorbitan oleate with high HLB and sorbitan monooleate with low HLB) at low emulsifier concentrations. Submicron emulsions prepared by the microfluidization process had smaller droplet diameters and exhibited less droplet diameter growth over time compared to high-speed homogenization. At emulsifier concentrations below 20% w/w, the droplet diameter or stability of the dispersed phase of the sub-micron emulsions prepared by either process was found to be dependent on the emulsifier content.

Drug Compounding↗

PubMed2006

Monolithic media in microfluidic devices for proteomics.

Considerable effort has been invested in the development of integrated microfluidic devices for fast and highly efficient proteomic studies. Among various fabrication techniques for the preparation of analytical components (separation columns, reactors, extractors, valves, etc.) in integrated microchips, in situ fabrication of monolithic media is receiving increasing attention. This is mainly due to the ease and simplicity of preparation of monolithic media and the availability of various precursors and chemistries. In addition, UV-initiated photopolymerization technique enables the incorporation of multiple analytical components into specified parts of a single microchip using photomasks. This review summarizes preparation methods for monolithic media and their application as microfluidic analytical components in microchips.

Microfluidic Analytical Techniques↗

PubMed2005

Bioactive heparin immobilized onto microfluidic channels in poly(dimethylsiloxane) results in hydrophilic surface properties.

A new composition of heparin coating for microfluidic systems made out of poly(dimethylsiloxane) (PDMS) was developed and evaluated. The coating that consists of a conditioning polyamine layer followed by two heparin/glutaraldehyde layers, resulted in channel surfaces with sufficient wettability to obtain flow of human normal plasma by capillary force alone. Hydrophilic channel walls are a desirable characteristic in microfluidic devices, since alternative pumping mechanisms must otherwise be included into the system. The immobilized heparin showed high antithrombin-binding capacity and a low degree of blood-material interaction. Plasma in contact with heparin-coated PDMS formed no detectable fibrin in a spectrophotometric assay by which plasma in contact with non-treated PDMS showed complete coagulation. The quartz crystal microbalance technique with energy dissipation monitoring (QCM-D) was utilized to obtain detailed information regarding adsorption kinetics and structural properties of the different layers composing the heparin coating.

Blood Flow Velocity↗

PubMed2004

A reusable microfluidic plate with alternate-choice architecture for assessing growth preference in tissue culture.

We present the design of a chamber to evaluate in vitro how species and concentrations of soluble molecules control features of cell growth-potentially including cell proliferation, cell motility, process extension, and process termination. We have created a reusable cell culture plate that integrates a microfluidic media delivery network with standard cell culture environment. The microfluidic network delivers a stream of cell culture media with a step-like concentration gradient down a 50-100 microm wide microchannel called the presentation region. Migrating cells or growing cell processes freely choose between the two distinct chemical environments in the presentation region, but they are forced to exclusively choose either one environment or the other when they grow past a physical barrier acting as a decision point. Our fabrication technique requires little specialized equipment, and can be carried out in approximately 4 days per plate. We demonstrate the effectiveness of our plates as neurites from spiral ganglion explants preferentially grow in media containing neurotrophin-3 (NT-3) as opposed to media without NT-3. Our design could be used without modification to study dissociated cell responses to soluble growth cues, and for behavioral screening of small motile organisms.

Animals↗

PubMed2006

Rapid quantification of bacterial cells in potable water using a simplified microfluidic device.

A simplified microfluidic device for quantification of bacteria in potable water was fabricated and examined. Comparisons of counts of Escherichia coli by the microfluidic system and by epifluorescence microscopy closely correlated (r2=0.99). Bacteria in natural mineral water and in purified household tap water were accurately enumerated by using this system within 15 min after fluorescent staining.

Colony Count, Microbial↗

PubMed2004

Quantitative microfluidic separation of DNA in self-assembled magnetic matrixes.

We present an experimental study of the microfluidic electrophoresis of long DNA in self-assembling matrixes of magnetic bead columns. Results are presented for the rapid separation of lambda-phage, 2lambda-DNA, and bacteriophage T4 DNA, where separation resolutions greater than 2 between lambda and T4 are achieved in times as short as 150 s. The use of a computer-piloted flow control system and injection results in high reproducibility between separations. We compare the experimentally measured mobility and dispersion with an exactly solvable lattice Monte Carlo model. The theory predicts that the mean velocity scales linearly with the field, the band broadening scales with the inverse of the field, and the resolution is independent of the field for intermediate fields-all of which are in accord with the experimental results. Moreover, reasonable quantitative agreement is achieved for band broadening for longer DNA (2lambda and T4) when the average postengagement time is measured experimentally. This work demonstrates the possibility of achieving fast microfluidic separation of large DNA on a routine basis.

DNA↗

PubMed2004

Formation of droplets of alternating composition in microfluidic channels and applications to indexing of concentrations in droplet-based assays.

For screening the conditions for a reaction by using droplets (or plugs) as microreactors, the composition of the droplets must be indexed. Indexing here refers to measuring the concentration of a solute by addition of a marker, either internal or external. Indexing may be performed by forming droplet pairs, where in each pair the first droplet is used to conduct the reaction, and the second droplet is used to index the composition of the first droplet. This paper characterizes a method for creating droplet pairs by generating alternating droplets, of two sets of aqueous solutions in a flow of immiscible carrier fluid within PDMS and glass microfluidic channels. The paper also demonstrates that the technique can be used to index the composition of the droplets, and this application is illustrated by screening conditions of protein crystallization. The fluid properties required to form the steady flow of the alternating droplets in a microchannel were characterized as a function of the capillary number Ca and water fraction. Four regimes were observed. At the lowest values of Ca, the droplets of the two streams coalesced; at intermediate values of Ca the alternating droplets formed reliably. At even higher values of Ca, shear forces dominated and caused formation of droplets that were smaller than the cross-sectional dimension of the channel; at the highest values of Ca, coflowing laminar streams of the two immiscible fluids formed. In addition to screening of protein crystallization conditions, understanding of the fluid flow in this system may extend this indexing approach to other chemical and biological assays performed on a microfluidic chip.

Calcium↗

PubMed2006

Parallel picoliter rt-PCR assays using microfluidics.

The development of microfluidic tools for high-throughput nucleic acid analysis has become a burgeoning area of research in the post-genome era. Here, we have developed a microfluidic chip to perform 72 parallel 450-pL RT-PCRs. We took advantage of Taqman hydrolysis probe chemistry to detect RNA templates as low as 34 copies. The device and method presented here may enable highly parallel single cell gene expression analysis.

Humans↗

PubMed2006

Continuous flow microfluidic demixing of electrolytes by induced charge electrokinetics in structured electrode arrays.

A continuous flow microfluidic demixing process is realized. It utilizes high external electrical fields that are applied over electrically floating noble metal electrodes in an otherwise straight microchannel. The process converts axial electrical potential gradients into lateral molecular selective transport via a structure oriented ensemble of numerous electrodes. While the individual electrodes locally modify the electrolyte distribution by nonlinear electrokinetic effects and concentration polarization, the directed orientation of the electrode array combines the individual polarization zones to a dedicated molecular enrichment against the generated concentration gradient. A homogeneously concentrated electrolyte can be separated into arbitrarily shaped laminae of increased and depleted concentration by the presented microfluidic demixer.

Electrodes↗

PubMedDate not supplied

Cell detachment model for an antibody-based microfluidic cancer screening system.

We consider cells bound to the floor of a microfluidic channel and present a model of their flow-induced detachment. We approximate hydrodynamic force and cell elastic response using static finite-element simulation of a single cell. Detachment is assumed to occur when hydrodynamic and adhesive forces are roughly equal. The result is extended to multiple cells at the device level using a sigmoidal curve fit. The model is applied to a microfluidic cancer-screening device that discriminates between normal epithelial cells and cells infected with human papillomavirus (HPV), on the basis of increased expression of the transmembrane protein alpha6 integrin in the latter. Here, the cells to be tested are bound to a microchannel floor coated with anti alpha6 integrin antibodies. In an appropriate flow rate range, normal cells are washed away while HPV-infected cells remain bound. The model allows interpolation between data points to choose the optimal flow rate and provides insight into interaction of cell mechanical properties and the flow-induced detachment mechanism. Notably, the results suggest a significant influence of cell elastic response on detachment.

Antibodies↗

PubMed2006

Highly reproducible method of planar lipid bilayer reconstitution in polymethyl methacrylate microfluidic chip.

We developed a highly reproducible method for planar lipid bilayer reconstitution using a microfluidic system made of a polymethyl methacrylate (PMMA) plastic substrate. Planar lipid bilayers are formed at apertures, 100 microm in diameter, by flowing lipid solution and buffer alternately into an integrated microfluidic channel. Since the amount and distribution of the lipid solution at the aperture determines the state of the lipid bilayer, controlling them precisely is crucial. We designed the geometry of the fluidic system so that a constant amount of lipid solution is distributed at the aperture. Then, the layer of lipid solution was thinned by applying an external pressure and finally became a bilayer when a pressure of 200-400 Pa was applied. The formation process can be simultaneously monitored with optical and electrical recordings. The maximum yield for bilayer formation was 90%. Using this technique, four lipid bilayers are formed simultaneously in a single chip. Finally, a channel current through gramicidin peptide ion channels was recorded to prove the compatibility of the chip with single molecule electrophysiology.

Lipid Bilayers↗

PubMed2006

Microfluidic approach for rapid multicomponent interfacial tensiometry.

We report a microfluidic instrument to rapidly measure the interfacial tension of multi-component immiscible liquids. The measurement principle rests upon the deformation and retraction dynamics of drops under extensional flow and was implemented for the first time in microfluidics (S. D. Hudson et al., Appl. Phys. Lett., 2005, 87, 081905 (ref. )). Here we describe in detail the instrument design and physics and extend this principle to investigate multicomponent mixtures, specifically two-component drops of adjustable composition. This approach provides fast real-time sigma measurements (on the order of 1 s), the possibility of rapidly adjusting drop composition and utilizes small sample volumes (approximately 10 microL). The tensiometer operation is illustrated with water drops and binary drops (water/ethylene glycol mixtures) in silicone oils. The technique should be particularly valuable for high-throughput characterization of complex fluids.

Equipment Design↗

PubMed2006

Pumpless, selective docking of yeast cells inside a microfluidic channel induced by receding meniscus.

We present a simple cell docking method induced by receding meniscus to capture non-adherent yeast cells onto microwells inside a microfluidic channel. Microwells were fabricated either by capillary moulding of UV curable polyurethane acrylate (PUA) onto glass substrate or direct replica moulding of poly(dimethyl siloxane) (PDMS). A cell suspension of the budding yeast, Saccharomyces cerevisiae, was introduced into the microfluidic channel by surface tension driven capillary flow and a receding meniscus was subsequently generated by evaporation. As the meniscus progressed, one to multiple yeast cells were spontaneously captured onto microwells by lateral capillary force created at the bottom of the meniscus. Using this cell-based platform, we observed the response of yeast cells upon stimulation by a mating pheromone (alpha-factor) by monitoring the expression of green fluorescent protein (GFP) with time. It was observed that alpha-factor triggered the expression of GFP at 60 min after stimulation and the fluorescence intensity was sustained for an additional 60 min without changes.

Cells, Immobilized↗

PubMed2022

Multiplexed microfluidic chip for cell co-culture.

Paracrine signaling is challenging to study in vitro, as conventional culture tools dilute soluble factors and offer little to no spatiotemporal control over signaling. Microfluidic chips offer potential to address both of these issues. However, few solutions offer both control over onset and duration of cell-cell communication, and high throughput. We have developed a microfluidic chip designed to culture cells in adjacent chambers, separated by valves to selectively allow or prevent exchange of paracrine signals. The chip features 16 fluidic inputs and 128 individually-addressable chambers arranged in 32 sets of 4 chambers. Media can be continuously perfused or delivered by diffusion, which we model under different culture conditions to ensure normal cell viability. Immunocytochemistry assays can be performed in the chip, which we modeled and fine-tuned to reduce total assay time to 1 h. Finally, we validate the use of the chip for co-culture studies by showing that HEK293Ta cells respond to signals secreted by RAW 264.7 immune cells in adjacent chambers, only when the valve between the chambers is opened.

Microfluidics↗

PubMed2004

Computerized microfluidic cell culture using elastomeric channels and Braille displays.

Computer-controlled microfluidics would advance many types of cellular assays and microscale tissue engineering studies wherever spatiotemporal changes in fluidics need to be defined. However, this goal has been elusive because of the limited availability of integrated, programmable pumps and valves. This paper demonstrates how a refreshable Braille display, with its grid of 320 vertically moving pins, can power integrated pumps and valves through localized deformations of channel networks within elastic silicone rubber. The resulting computerized fluidic control is able to switch among: (i) rapid and efficient mixing between streams, (ii) multiple laminar flows with minimal mixing between streams, and (iii) segmented plug-flow of immiscible fluids within the same channel architecture. The same control method is used to precisely seed cells, compartmentalize them into distinct subpopulations through channel reconfiguration, and culture each cell subpopulation for up to 3 weeks under perfusion. These reliable microscale cell cultures showed gradients of cellular behavior from C2C12 myoblasts along channel lengths, as well as differences in cell density of undifferentiated myoblasts and differentiation patterns, both programmable through different flow rates of serum-containing media. This technology will allow future microscale tissue or cell studies to be more accessible, especially for high-throughput, complex, and long-term experiments. The microfluidic actuation method described is versatile and computer programmable, yet simple, well packaged, and portable enough for personal use.

Audiovisual Aids↗

PubMed2007

Coding/decoding and reversibility of droplet trains in microfluidic networks.

Droplets of one liquid suspended in a second, immiscible liquid move through a microfluidic device in which a channel splits into two branches that reconnect downstream. The droplets choose a path based on the number of droplets that occupy each branch. The interaction among droplets in the channels results in complex sequences of path selection. The linearity of the flow through the microchannels, however, ensures that the behavior of the system can be reversed. This reversibility makes it possible to encrypt and decrypt signals coded in the intervals between droplets. The encoding/decoding device is a functional microfluidic system that requires droplets to navigate a network in a precise manner without the use of valves, switches, or other means of external control.

Alkanes↗