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Microfluidic polymer chip integrated with an ISFET detector for cationic surfactant assay in dental rinses.

A cationic surfactant ion-selective field-effect transistor (cationic surfactant-ISFET) has been developed based on the tetraphenylborate derivative known as sodium tetrakis[3,5-bis(trifluoromethyl)phenyl]borate. The cationic surfactant-ISFET shows an almost Nernstian response to tetradecyldimethylbenzylammonium chloride (Zephiramine) over a concentration range between 1.0 x 10(-6) M and 1.0 x 10(-3) M, with a slope of 58.5 +/- 1.7 mV/decade. The cationic surfactant-ISFET can be used over a range of pH values, between pH 3 and 9. The cationic surfactant-ISFET shows excellent selectivity for Zephiramine over small inorganic cations, but shows similar selectivity for other cationic surfactants, such as hexadecyltrimethylammonium and stearyltrimethylammonium ions. A microfluidic polymer chip was integrated with the cationic surfactant-ISFET, and this was fabricated using polystyrene plates and stainless wires as a template for the channel. Cationic surfactant-ISFETs used in a batch system and microchips integrated with cationic surfactant-ISFETs showed very similar performance in terms of low detection limits, slope sensitivity and the stability of the potential response. The microfluidic polymer chip was then applied to the determination of cationic surfactants in dental rinses.

Calibration↗

[Developments in microfluidic spectrometry].

A review is presented on the development of spectrometric detection in microfluidic chemical analysis systems. A brief introduction of microfluidic analysis and future perspectives of the technique are also discussed.

Animals↗

Microfluidics: an opportunity for trend-setting drug delivery.

The past has demonstrated that when working with microfluidics only the following scenario is likely to lead to successful system development: a high level of component integration at an early development stage with an application-specific, simple (reliable) design, and a high interdisciplinary level of know-how over a broad range of technologies, pharmacology and medicine. The mere application of existing nonspecifically designed or outsourced microfluidic components is likely to fail because the system behaviour in the microworld is far different from that in the macroworld.

Drug Delivery Systems↗

[Design of a microfluidic immunoassay system based on superparamagnetic microbeads].

We use direct-write laser micromachining technology to fabricate the microfluidic chip, and to establish a microfluidic chemiluminescence immunoassay system based on superparamagnetic microbeads, for detecting alpha- fetoprotein (AFP). The AFP analysis can be completed in 20 minutes with 5 microl sample and 5 microl reagent, and there is a good linear correlation in the range of 1-800 ng/ml.

Equipment Design↗

Polymer microfabrication methods for microfluidic analytical applications.

A growing number of microsystem technology (MST) applications, particularly in the field of microfluidics with its applications in the life sciences, have a need for novel fabrication methods which account for substrates other than silicon or glass. We present in this paper an overview of existing polymer microfabrication technologies for microfluidic applications, namely replication methods such as hot embossing, injection molding and casting, and the technologies necessary to fabricate the molding masters. In addition, techniques such as laser ablation and layering techniques are examined. Methods for bonding and dicing of polymer materials, which are necessary for complete systems, are evaluated.

Chemistry Techniques, Analytical↗

New approaches for fabrication of microfluidic capillary electrophoresis devices with on-chip conductivity detection.

In practice, microfluidic systems are based on the principles of capillary electrophoresis (CE), for a large part due to the simplicity of electroosmotic pumping. In this contribution, a universal conductivity detector is presented that allows detection of charged species down to the microM level. Additionally, powderblasting is presented as a novel technique for direct etching of microfluidic networks. This method allows creation of features down to 50 microm with a total processing time (design to device) of less than one day. The performance of powderblasted devices with integrated conductivity detection is illustrated by the separation of lithium, sodium, and potassium ions and that of fumaric, malic, and citric acid.

Acids↗

A disposable poly(methylmethacrylate)-based microfluidic module for protein identification by nanoelectrospray ionization-tandem mass spectrometry.

The design, fabrication, and analytical use of a poly(methylmethacrylate) (PMMA)-based microfluidic module for nanoelectrospray ionization-tandem mass spectrometry (nano-ESI-MS/MS) were described. The microfluidic module can be mass-produced at low costs and used as a disposable device to generate nano-ESI-MS/MS signals for protein identification from low amounts of protein samples. Compared with commercially available nanospray capillary tips, the module gave comparable signal quality and also offered advantages in convenience and easiness of operation, permitting repeated usage, and disposability.

Animals↗

Multiplexed enzyme assays in capillary electrophoretic single-use microfluidic devices.

We describe a method of performing multiple enzyme assays in a single reaction vessel. The resolving power of capillary electrophoresis enables several enzyme assays to be analyzed at high speed in microfluidic arrays. Multiplexed measurement can increase throughput significantly without requiring highly dense microfluidic arrays. Enzyme assays in a multiplexed format for selected kinases in this work show essentially identical performance to assays performed individually. This establishes an approach for screening one compound against multiple enzyme targets simultaneously. Another potential application for performing multiplexed enzyme assay is to study protein-protein (especially enzyme-enzyme) interaction by monitoring the enzymatic activity changes.

Amino Acid Sequence↗

A world-to-chip socket for microfluidic prototype development.

We report a prototype for a standard connector between a microfluidic chip and the macroworld. This prototype is the first to demonstrate a fully functioning socket for a microchip to access the outside world by means of fluids, data, and energy supply, as well as providing process visibility. It has 20 channels for the input and output of liquids or gases, as well as compressed air or vacuum lines for pneumatic power lines. It also contains 42 pins for electrical signals and power. All these connections were designed in a planar configuration with linear orthogonal arrays. The vertical space was opened for optical measurement and evaluation. The die (29.1 mm x 27.5 mm x 0.9 mm) can be easily mounted and dismounted from the socket. No adhesives or solders are used at any contact points. The pressure limit for the connection of working fluids was 0.2 MPa and the current limit for the electrical connections was 1 A. This socket supports both serial and parallel processing applications. It exhibits great potential for developing microfluidic systems efficiently.

Electrochemistry↗

Miniaturized capillary isoelectric focusing in plastic microfluidic devices.

We report the demonstration of miniaturized capillary isoelectric focusing (CIEF) in plastic microfluidic devices. Conventional CIEF technique was adapted to the microfluidic devices to separate proteins and to detect protein-protein interactions. Both acidic and basic proteins with isoelectric points (pI) ranging from 5.4 to 11.0 were rapidly focused, mobilized, and detected in a 1.2 cm long channel (50 microm deep x 120 microm wide) with a total analysis time of 150 s. In a device with a focusing distance of 4.7 cm, the separation efficiency for a basic protein, lysozyme, was achieved as high as 1.5 x 10(5) plates, corresponding to 3.2 million plates per meter. We also experimentally confirmed that IEF resolution is essentially independent of focusing length when the applied voltage is kept the same and within a range that it does not cause Joule heating. Further, we demonstrated the use of miniaturized CIEF to study the interactions between two pairs of proteins, immunoglobulin G (IgG) with protein G and anti-six histidine (anti-6xHis) with 6xHis-tagged green fluorescent protein (GFP). Using this approach, protein-protein interactions can be detected for as little as 50 fmol of protein. We believe miniaturized CIEF is useful for studying protein-protein interactions when there is a difference in pI between a protein-protein complex and its constitutent proteins.

Electrophoresis, Capillary↗

Dynamic analyte introduction and focusing in plastic microfluidic devices for proteomic analysis.

Isoelectric focusing (IEF) separations, in general, involve the use of the entire channel filled with a solution mixture containing protein/peptide analytes and carrier ampholytes for the creation of a pH gradient. Thus, the preparative capabilities of IEF are inherently greater than most microfluidics-based electrokinetic separation techniques. To further increase sample loading and therefore the concentrations of focused analytes, a dynamic approach, which is based on electrokinetic injection of proteins/peptides from solution reservoirs, is demonstrated in this study. The proteins/peptides continuously migrate into the plastic microchannel and encounter a pH gradient established by carrier ampholytes originally present in the channel for focusing and separation. Dynamic sample introduction and analyte focusing in plastic microfluidic devices can be directly controlled by various electrokinetic conditions, including the injection time and the applied electric field strength. Differences in the sample loading are contributed by electrokinetic injection bias and are affected by the individual analyte's electrophoretic mobility. Under the influence of 30 min electrokinetic injection at constant electric field strength of 500 V/cm, the sample loading is enhanced by approximately 10-100 fold in comparison with conventional IEF.

Animals↗

A novel microfluidic mixer utilizing electrokinetic driving forces under low switching frequency.

This paper presents a novel technique in which low-frequency periodic electrokinetic driving forces are utilized to mix electrolytic fluid samples rapidly and efficiently in a double-T-form microfluidic mixer. Without using any additional equipment to induce flow perturbations, only a single high-voltage power source is required for simultaneously driving and mixing the sample fluids which results in a simple and low-cost system for the mixing purpose. The effectiveness of the mixer as a function of the applied electric field and the periodic switching frequency is characterized by the intensity distribution calculated downstream from the mixing zone. The present numerical and experimental results confirm that the proposed double-T-form micromixer has excellent mixing capabilities. The mixing efficiency can be as high as 95% within a mixing length of 1000 microm downstream from the secondary T-junction when a 100 V/cm driving electric field strength and a 2 Hz periodic switching frequency are applied. The results reveal that the optimal switching frequency depends upon the magnitude of the main applied electrical field. The rapid double-T-form microfluidic mixer using the periodic driving voltage switching model proposed in this study has considerable potential for use in lab-on-a-chip systems.

Electrophoresis, Microchip↗

Chiral separations on multichannel microfluidic chips.

Chiral separations of FITC-labeled basic drugs on multichannel microfluidic chips with LIF detector were investigated. A preliminary screening procedure for seven neutral CDs was performed under optimized conditions for chiral separations of three FITC-labeled drugs (baclofen, norfenefrine, and tocainide) on a mono-channel microfluidic chip. According to the results of screening, FITC-baclofen and FITC-norfenefrine as well as two chiral selectors including gamma-CD and dimethyl-beta-CD (DM-beta-CD) were selected as models to perform chiral separations on a two-channel chip. FITC-baclofen enantiomers were separated completely by gamma-CD in one channel, while resolution of FITC-norfenefrine enantiomers was achieved by DM-beta-CD in the other channel in the same run. Furthermore, the feasibility of using one chiral selector to separate multiple chiral samples was studied on a four-channel chip. These results show that multichannel chip has a potential for chiral high-throughput screening.

Baclofen↗

A microfluidic device for measuring cellular membrane potential.

Recent developments in microfluidics have enabled the design of a lab-on-a-chip system capable of measuring cellular membrane potential. The chip accesses liquid samples sequentially by sipping from a microplate through a capillary, mixes the samples with cells flowing through a microchannel, contacts the cells with potential-sensitive dyes, and reads out cellular responses using fluorescence detection. The rate of cellular uptake of membrane-permeable, ionic fluorophores by THP-1 cells was found to depend strongly on membrane potential. The ratio of the fluorescence of the anionic dye DiBAC(4)(3) and the cationic dye Syto 62 taken up by cells was found to double for every 33 mV change in membrane potential. The utility of this approach was demonstrated by assaying ion channel activity in human T lymphocytes. Because of the high sensitivity, low cellular and reagent consumption, and high data quality obtained with the microfluidic device, the lab-on-a-chip system should be widely applicable in high-throughput screening and functional genomics studies.

Barbiturates↗

Theoretical studies of microfluidic dispensing processes.

The understanding of electrokinetic transport phenomena in microfluidic dispensers, an important component of biochips, is very important for designing and controlling biochips. A theoretical model to study the electrokinetic transport processes in microfluidic dispensers was developed in the work to study the controlling parameters for the dispensing process. The electrical field, the flow field, and the concentration field during dispensing processes were obtained by solving this theoretical model numerically. The effects of the electroosmotic mobility of the buffer solution, the diffusion coefficient and the electrophoretic mobility of the sample, the applied electrical field strength, and the channel size on the dispensing process are examined in this paper. The investigations show that optimal controlling parameter values can be found by using this model for dispensing any desired amount of the sample.

Journal Article↗

Multiport flow-control system for lab-on-a-chip microfluidic devices.

A multiport system suitable for pressure control on a lab-on-a-chip microfluidic device is described. An algorithm and a strategy for calculating pressures were developed to control the flow from multiple reservoirs for the microfluidic devices. Dye mixing and enzyme assay titration experiments were performed using pressure-driven flow only. Results show a good linear response over two orders of dynamic range.

Algorithms↗

Hard and soft micromachining for BioMEMS: review of techniques and examples of applications in microfluidics and drug delivery.

Recent development in microfabrication (micromachining, microelectromechanical systems, MEMS) permits the integration of hard and soft structures, and enables the design of controllable microfluidic systems, which may be applied to drug delivery. In this paper, we present a tutorial review of both classical "hard" and more recent "soft" micromachining techniques. We then provide examples where these techniques are combined to produce hydrogel-based microfluidic control systems. The most complex of these systems utilizes a very small hydrogel based on phenylboronic acid to control the flow of an insulin solution in response to changes in glucose concentration.

Drug Delivery Systems↗

Microfluidic light scattering as a tool to study the structure of aqueous polymer solutions.

A small-angle light scattering (SALS) apparatus, coupled with a specially designed microfluidic device is shown to monitor the formation and subsequent size distribution of giant multilamellar vesicles of a diblock copolymer in aqueous solution. The closed-face design, fabricated between glass slides using a UV-curable optical adhesive, incorporates multiple inlets, a mixing system, and a viewing window to perform on-line SALS. The mixing of each component is tested using polystyrene latex microspheres. Vesicles of the block copolymer, EO6BO11 in aqueous solution are formed on the SALS chip and the pair distance distribution function determined using an inverse Fourier transformation of the scattered intensity to quantify the population and distribution for a range of vesicle sizes. These experiments provide demonstrations of how SALS on a microfluidic device can be used as a rapid screening tool to optimize processing conditions for a range of polymer solutions.

Coloring Agents↗