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Results for “Microfluidics”

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At least 829 records · Page 46Linked to original sources

PubMed2004

Systematic investigation of protein phase behavior with a microfluidic formulator.

We demonstrated a microfluidic device for rapidly generating complex mixtures of 32 stock reagents in a 5-nl reactor. This "formulation chip" is fully automated and allows thousands of experiments to be performed in a single day with minimal reagent consumption. It was applied to systematically study the phase behavior of the protein xylanase over a large and complex chemical space. For each chemical formulation that demonstrated a pronounced effect on solubility, the protein phase behavior was completely mapped in the chip, generating a set of empirical phase diagrams. This ab initio phase information was used to devise a rational crystallization screen that resulted in 72-fold improvement in successful crystallization hits compared with conventional sparse matrix screens. This formulations tool allows a physics-based approach to protein crystallization that may prove useful in structural genomics efforts.

Biophysical Phenomena↗

PubMed2006

High-speed microfluidic differential manometer for cellular-scale hydrodynamics.

We propose a broadly applicable high-speed microfluidic approach for measuring dynamical pressure-drop variations along a micrometer-sized channel and illustrate the potential of the technique by presenting measurements of the additional pressure drop produced at the scale of individual flowing cells. The influence of drug-modified mechanical properties of the cell membrane is shown. Finally, single hemolysis events during flow are recorded simultaneously with the critical pressure drop for the rupture of the membrane. This scale-independent measurement approach can be applied to any dynamical process or event that changes the hydrodynamic resistance of micro- or nanochannels.

Calibration↗

PubMed2006

Designing a nano-interface in a microfluidic chip to probe living cells: challenges and perspectives.

Nanotechnology-based materials are beginning to emerge as promising platforms for biomedical analysis, but measurement and control at the cell-chip interface remain challenging. This idea served as the basis for discussion in a focus group at the recent National Academies Keck Futures Initiative. In this Perspective, we first outline recent advances and limitations in measuring nanoscale mechanical, biochemical, and electrical interactions at the interface between biomaterials and living cells. Second, we present emerging experimental and conceptual platforms for probing living cells with nanotechnology-based tools in a microfluidic chip. Finally, we explore future directions and critical needs for engineering the cell-chip interface to create an integrated system capable of high-resolution analysis and control of cellular physiology.

Biomedical Engineering↗

PubMed2006

A high-throughput microfluidic assay for SH2 domain-containing inositol 5-phosphatase 2.

SH2 domain-containing inositol 5-phosphatase 2 (SHIP2) is a potential drug target for the treatment of type 2 diabetes. This enzyme serves as a negative regulator of insulin-mediated signal transduction by catalyzing the dephosphorylation of the second messenger lipid molecule phosphatidylinositol 3,4,5-triphosphate. Traditionally, assays for phosphoinositide phosphatases such as SHIP2 have relied on radiolabeled phosphatidylinositol-containing lipid membranes and chromatographic separation of labeled phospholipid substrate from product by thin-layer chromatography. We have expressed and purified catalytically active phosphatase domain constructs of SHIP2 from Escherichia coli and developed a sensitive and antibody- or binding protein-independent assay for SHIP2 amenable to high-throughput screening of phosphoinositide phosphatases or phosphoinositide kinases. This microfluidic assay, with Z' values approximately 0.8, is based upon the difference in mobility within an electric field between a fluorophore-labeled phosphatidylinositol 3,4,5-triphosphate substrate and the corresponding 3,4-bisphosphate product. High-throughput screening of a 91,060-member compound library in 384-well format resulted in the identification of SHIP2 inhibitors.

Chromatography, Thin Layer↗

PubMed2006

Purification and preconcentration of genomic DNA from whole cell lysates using photoactivated polycarbonate (PPC) microfluidic chips.

We discuss the use of a photoactivated polycarbonate (PPC) microfluidic chip for the solid-phase, reversible immobilization (SPRI) and purification of genomic DNA (gDNA) from whole cell lysates. The surface of polycarbonate was activated by UV radiation resulting in a photo-oxidation reaction, which produced a channel surface containing carboxylate groups. The gDNA was selectively captured on this photoactivated surface in an immobilization buffer, which consisted of 3% polyethylene glycol, 0.4 M NaCl and 70% ethanol. The methodology reported herein is similar to conventional SPRI in that surface-confined carboxylate groups are used for the selective immobilization of DNA; however, no magnetic beads or a magnetic field are required. As observed by UV spectroscopy, a load of approximately 7.6 +/- 1.6 microg/ml of gDNA was immobilized onto the PPC bed. The recovery of DNA following purification was estimated to be 85 +/- 5%. The immobilization and purification assay using this PPC microchip could be performed within approximately 25 min as follows: (i) DNA immobilization approximately 6 min, (ii) chip washout with ethanol 10 min, and (iii) drying and gDNA desorption approximately 6 min. The PPC microchip could also be used for subsequent assays with no substantial loss in recovery, no observable carryover and no need for 'reactivation' of the PC surface with UV light.

Buffers↗

PubMed2004

Engineering the morphology and electrophysiological parameters of cultured neurons by microfluidic surface patterning.

The ability to control the orientation, morphology, and electrophysiological characteristics of neurons in culture allows the construction of neural circuits with defined physiological properties. Using microfluidic protein deposition onto chemically modified glass, we achieve the controlled growth of Aplysia neurons on geometrical patterns of poly-L-lysine and collagen IV, surrounded by nonadhesive regions of bovine albumin. We investigate the parameters essential for forming functional neuronal networks, the morphology, biochemistry, and electrophysiology under engineered cell culture conditions. We demonstrate that not only the orientation of neurite extension but also the number of primary neurites originating from the cell soma, their length, and branching pattern depend on the spatial constraints presented by the size and shape of the adhesion region on the patterned substrate. In addition, the physicochemical properties of the support layer influence the electrical activity of the cultured neurons. Substrate-dependent changes in the amplitude and in the dynamic parameters of the action potential cause decreased spike broadening in patterned neurons, which reflects changes in the number or functioning of active membrane ion channels. In contrast to morphology and electrophysiology, the neuropeptide content, as determined by mass spectrometry of individual patterned neurons, is not affected by the growth on patterned surfaces. Our results suggest that the morphological and electrophysiological parameters of neurons can be predictably altered/engineered by modulation of the chemical, physical, and topographical features of culture substrates. We also demonstrate that a full suite of techniques is required for functional characterization of neurons on engineered substrates.

Action Potentials↗

PubMed2005

Bubble dispenser in microfluidic devices.

This Brief Report presents experimental and computational results on bubble formation in microfluidic devices. Bubbles are generated at the right-angle intersection of four identical square microchannels. When the pressure gradient generated by the liquid flow dominates the pressure gradient generated by gas flow, the length of the produced confined bubbles follows a law based on the channel size and fluid volume fraction. This bubble production technique was used to produce monodisperse aqueous foam in two-dimensional and three-dimensional microchannels.

Computer Simulation↗

PubMed2006

DNA electrophoresis in microfluidic post arrays under moderate electric fields.

The dynamics of long DNA moving through microfluidic arrays of micron-sized posts under a moderate electric field are modeled by a Scher-Lax continuous-time random walk. The microscale model consists of a repetitive sequence of three steps: (i) collision with the post and extension into two arms, (ii) electric-field-driven unhooking from the post, and (iii) uniform translation until the next collision. The model features two random variables: the initial offset between the two arms of the polymer during a given collision and the distance traveled between collisions. For experimentally realistic values of the electric field strength and DNA molecular weight, scaling laws indicate that the chain will generally be in a stem-flower conformation when unhooking from the post. Compared to a taut-chain model at the same field strength, the stem-flower conformation reduces the time engaged with the post and increases the collision frequency. Analytical expressions for the mean velocity and dispersivity are derived as a function of the post density, post spacing, free-solution mobility, Kuhn length, and sequence length. The incomplete extension of the chain does not strongly affect the mean velocity, but tends to increase the dispersivity relative to a taut chain. As a result, the separation resolution decreases as the field decreases for a moderate field, in agreement with experiments. The quantitative agreement between the model and experimental data is satisfactory, especially considering that the model contains no adjustable parameters.

DNA↗

PubMed2005

Non-markovian transport of DNA in microfluidic post arrays.

We present an analytically solvable model for the transport of long DNA through microfluidic arrays of posts. The motion is a repetitive three-part cycle: (i) collision with the post and extension of the arms; (ii) rope-over-pulley post disengagement; and (iii) a random period of uniform translation before the next collision. This cycle, inspired by geometration, is a nonseparable (Scher-Lax) continuous-time random walk on a lattice defined by the posts. Upon adopting a simple model for the transition probability density on the lattice, we analytically compute the mean DNA velocity and dispersivity in the long-time limit without any adjustable parameters. The results compare favorably with the limited amount of experimental data on separations in self-assembled arrays of magnetic beads. The Scher-Lax formalism provides a template for incorporating more sophisticated microscale models.

Bacteriophage T4↗

PubMed2005

Microfluidics of soft matter investigated by small-angle X-ray scattering.

The combination of X-ray microdiffraction and microfluidics is used to investigate the dynamic behaviour of soft materials. A microfocused X-ray beam enables the observation of the influence of droplet formation on the nanostructure of a smectic liquid crystal in water. Using a hydrodynamic focusing device, the evolution of the intercalation of DNA into multilamellar membranes can be studied. Owing to the elongational flow at the centre of this device, alignment of the material is induced which allows for an improved structural characterization. Furthermore, the influence of strain applied to these materials can be tested.

Animals↗

PubMed2004

A magnetocaloric pump for microfluidic applications.

A magnetocaloric pump provides a simple means of pumping fluid using only external thermal and magnetic fields. The principle, which can be traced back to the early work of Rosensweig, is straightforward. Magnetic materials tend to lose their magnetization as the temperature approaches the material's Curie point. Exposing a column of magnetic fluid to a uniform magnetic field coincident with a temperature gradient produces a pressure gradient in the magnetic fluid. As the fluid heats up, it loses its attraction to the magnetic field and is displaced by cooler fluid. The impact of such a phenomenon is obvious: fluid propulsion with no moving mechanical parts. Until recently, limitations in the magnetic and thermal properties of conventional materials severely limited practical operating pressure gradients. However, recent advancements in the design of metal substituted magnetite enable fine control over both the magnetic and thermal properties of magnetic nanoparticles, a key element in colloidal-based magnetic fluids (ferrofluids). This paper begins with a basic description of the process and previous limitations due to material properties. This is followed by a review of existing methods of synthesizing magnetic nanoparticles as well as an introduction to a new approach based on thermophilic metal-reducing bacteria. We compare two compounds and show, experimentally, significant variation in specific magnetic and thermal properties. We develop the constitutive thermal, magnetic, and fluid dynamic equations associated with a magnetocaloric pump and validate our finite-element model with a series of experiments. Preliminary results show a good match between the model and experiment as well as approximately an order of magnitude increase in the fluid flow rate over conventional magnetite-based ferrofluids operating below 80 degrees C. Finally, as a practical demonstration, we describe a novel application of this technology: pumping fluids at the "lab-on-a-chip" microfluidic scale.

Computer-Aided Design↗

PubMed2006

Simple detection of small amounts of Pseudomonas cells in milk by using a microfluidic device.

AIMS: Flow cytometry offers rapid and reliable analyses of bacteria in milk. However, a flow cytometer is relatively expensive and operation is rather complicated for an unskilled operator. We applied flow cytometry using a microfluidic device (on-chip flow cytometry) in detection of small amounts of milk-spoiling bacteria. METHODS AND RESULTS: Pseudomonas cells in milk were in situ hybridized with Cy5-labelled probe specific for Pseudomonas spp. under optimized condition. Numbers of Pseudomonas cells in the stationary phase and in the starved state determined by on-chip flow cytometry were compared with those determined by conventional plate counting, and on-chip flow cytometry detected targeted cells in milk that were undetectable as colony forming units(CFU) on Standards Methods Agar. CONCLUSIONS: The contamination in milk with fewer than 10 CFU ml(-1) of targeted cells in starved state was detectable with simple procedure (0.5 h milk-clearing, 1 h fixation, 2 h hybridization and 0.5 h on-chip flow cytometry following 12 h enrichment of cells). SIGNIFICANCE AND IMPACT OF THE STUDY: On-chip flow cytometry following fluorescence in situ hybridization could be applicable to simple detection of milk-spoiling bacteria.

Animals↗

PubMedDate not supplied

Vertical-cavity semiconductor devices for fluorescence spectroscopy in biochips and microfluidic platforms.

This paper describes the properties of vertical-cavity semiconductor devices designed to emit light when driven in forward bias mode and detect optical radiation at wavelengths longer than that of emission when driven in reverse bias mode. The study of this type of devices is motivated by the miniaturization and integration into a single unit of the three functions that a microfluorimeter has to perform, optical pumping, optical detection, and optical filtering of weak light sources. The devices produced can generate fluorescence with a low output power since their emission wavelength can be tuned with that of maximum absorption of the fluorescent dye. We demonstrate also that they can detect low power fluorescence generated in a small volume of concentrated solution of a commercial dye. These devices can find useful application in microanalytical systems such as microfluidic devices or optical biochips.

Biosensing Techniques↗

PubMed2005

Multistep synthesis of a radiolabeled imaging probe using integrated microfluidics.

Microreactor technology has shown potential for optimizing synthetic efficiency, particularly in preparing sensitive compounds. We achieved the synthesis of an [(18)F]fluoride-radiolabeled molecular imaging probe, 2-deoxy-2-[18F]fluoro-D-glucose ([18F]FDG), in an integrated microfluidic device. Five sequential processes-[18F]fluoride concentration, water evaporation, radiofluorination, solvent exchange, and hydrolytic deprotection-proceeded with high radio-chemical yield and purity and with shorter synthesis time relative to conventional automated synthesis. Multiple doses of [18F]FDG for positron emission tomography imaging studies in mice were prepared. These results, which constitute a proof of principle for automated multistep syntheses at the nanogram to microgram scale, could be generalized to a range of radiolabeled substrates.

Animals↗

PubMed2005

Rapid and simple quantification of bacterial cells by using a microfluidic device.

This study investigated a microfluidic chip-based system (on-chip flow cytometry) for quantification of bacteria both in culture and in environmental samples. Bacterial numbers determined by this technique were similar to those obtained by direct microscopic count. The time required for this on-chip flow cytometry was only 30 min per 6 samples.

Bacteriological Techniques↗

PubMed2005

Evanescent field-based optical fiber sensing device for measuring the refractive index of liquids in microfluidic channels.

We report a simple optical sensing device capable of measuring the refractive index of liquids propagating in microfluidic channels. The sensor is based on a single-mode optical fiber that is tapered to submicrometer dimensions and immersed in a transparent curable soft polymer. A channel for liquid analyte is created in the immediate vicinity of the taper waist. Light propagating through the tapered section of the fiber extends into the channel, making the optical loss in the system sensitive to the refractive-index difference between the polymer and the liquid. The fabrication process and testing of the prototype sensing devices are described. The sensor can operate both as a highly responsive on-off device and in the continuous measurement mode, with an estimated accuracy of refractive-index measurement of approximately 5 x 10(-4).

Complex Mixtures↗

PubMed2005

Laser-tweezer-controlled solid immersion microscopy in microfluidic systems.

We describe the creation and implementation of a near-field scanning solid immersion microscope that is specifically tailored for use in microfluidic systems. The microscope comprises a newly fabricated Weierstrass solid immersion lens (SIL), which is detached from its substrate and is free floating in the fluid, and a laser optical tweezer, which serves both as a trapping beam for alignment and positioning of the SIL and as a near-field scanning beam that images the sample through the SIL. A discussion of the SIL's fabrication method is presented along with experimental results that demonstrate the effectiveness of our microscope design.

Equipment Design↗

PubMed2004

Application of fluorescence correlation spectroscopy for velocity imaging in microfluidic devices.

In this paper we present and demonstrate a technique for mapping fluid flow rates in microfluidic systems with sub-micrometer resolution using confocal microscopy in conjunction with fluorescence correlation spectroscopy (FCS). Flow velocities ranging from approximately 50 microm/s to approximately 10 cm/s can be recorded using fluorescent polymer nanospheres as fluid motion tracers. Velocity profiles and images of the flow in poly(dimethylsiloxane)-glass microchannels are presented and analyzed. Using the method, velocity images along the horizontal (top view) and vertical planes within a microdevice can be obtained. This is, to our knowledge, the first report of FCS for producing velocity maps. The high-resolution velocity maps can be used to characterize and optimize microdevice performance and to validate simulation efforts.

Chemistry, Pharmaceutical↗