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At least 811 records · Page 45Linked to original sources
PubMed2001

Development of a new flow-through bulk optode for the determination of manganese(III).

A flow-through bulk optode based on the use of 1-(2-pyridylazo)-2-naphthol (PAN) immobilized in a plasticized poly(vinyl chloride) membrane entrapped in a cellulose support, in conjuntion with the flow injection analysis technique, is proposed for the determination of manganese(II). The calibration graph obtained at 570 nm was linear in the range 0.27-27.5 mg L(-1) (5 x 10(-6)-5 x 10(-4) M) Mn(II) with a detection limit of 0.18 mg L(-1). The coefficients of variation of the sensor response for 5.5 mg L(-1) of Mn(II) were +/-0.22% for consecutive measurements (n = 10), +/-0.48% between days (n = 5) and +/-0.38% between different membranes (n = 6). The sensor was readily regenerated with the carrier acetic acid/acetate buffer of pH 4.5. The method was applied to the determination of manganese in steels, waters and lemon tree leaves.

Journal Article↗
PubMed2001

Analytical methodologies for aluminium speciation in environmental and biological samples--a review.

It is recognized that aluminium (Al) is a potential environmental hazard. Acidic deposition has been linked to increased Al concentrations in natural waters. Elevated levels of Al might have serious consequences for biological communities. Of particular interest is the speciation of Al in aquatic environments, because Al toxicity depends on its forms and concentrations. In this paper, advances in analytical methodologies for Al speciation in environmental and biological samples during the past five years are reviewed. Concerns about the specific problems of Al speciation and highlights of some important methods are elucidated in sections devoted to hybrid techniques (HPLC or FPLC coupled with ET-AAS, ICP-AES, or ICP-MS), flow-injection analysis (FIA), nuclear magnetic resonance (27Al NMR), electrochemical analysis, and computer simulation. More than 130 references are cited.

Aluminum↗
PubMed2001

Multi-layer microfluidic glass chips for microanalytical applications.

A new, versatile architecture is presented for microfluidic devices made entirely from glass, for use with reagents which would prove highly corrosive for silicon. Chips consist of three layers of glass wafers bonded together by fusion bonding. On the inside wafer faces a network of microfluidic channels is created by photolithography and wet chemical etching. Low dead-volume fluidic connections between the layers are fabricated by spark-assisted etching (SAE), a computer numerical controlled (CNC)-like machining technique new to microfluidic system fabrication. This method is also used to form a vertical, long path-length, optical cuvette through the middle wafer for optical absorbance detection of low-concentration compounds. Advantages of this technique compared with other, more standard, methods are discussed. When the new glass-based device for flow-injection analysis of ammonia was compared with our first-generation chips based on silicon micromachining, concentration sensitivity was higher, because of the longer path-length of the optical cuvette. The dependence of dispersion on velocity profile and on channel cross-sectional geometry is discussed. The rapid implementation of the devices for an organic synthesis reaction, the Wittig reaction, is also briefly described.

Journal Article↗
PubMed1996

Determination of salicylate in beverages and cosmetics by use of an amperometric biosensor.

A fast and selective enzymatic method for the determination of salicylate in beverages and cosmetics has been developed. The enzyme salicylate hydroxylase was immobilised covalently onto a glassy carbon working electrode of a wall-jet cell coupled with a flow-injection analysis system. The salicylate is enzymatically converted to catechol, which can be detected amperometrically on the glassy carbon electrode at +0.45 V. The response of the biosensor is linearly proportional to the concentration of salicylate between 725 nmol/l and 700 micromol/l. A high sample throughput (60 h(-1)) is possible, and the biosensor is stable for more than three months. Sample pretreatment for beverages and hair lotions is easy and fast. For creams, an extraction of salicylate is necessary. Relative standard deviations are less than 5.5% and the recoveries are between 95 and 105%.

Journal Article↗
PubMed1996

High volumetric yields of functional dimeric miniantibodies in Escherichia coli, using an optimized expression vector and high-cell-density fermentation under non-limited growth conditions.

Functional bivalent miniantibodies, directed against the epidermal growth factor receptor, accumulated to more than 3 gl-1 in high-cell-density cultures of Escherichia coli RV308(pHKK) on a pilot scale. The miniantibodies consist of scFv fragments with a C-terminal hinge followed by a helix-turn-helix motif, which homodimerizes in vivo. The improved expression vector pHKK is characterized by the hoklsok suicide system, improving plasmid maintenance, and the inducible lac pl o promoter system with the very strong T7g10 Shine-Dalgarno sequence. The expression unit is flanked by terminators. The prototrophic RV308 cells were cultivated in glucose mineral salt medium and reached a cell density of 145 g dry biomass l-1 after 33 h. After induction, growth continued almost unchanged for a further 4 h with concomitant miniantibody formation. In the fedbatch phase, the concentration of glucose was kept almost constant at the physiological level of approximately 1.5 gl-1, using on-line flow injection analysis for control. Surprisingly, E. coli RV308(pHKK) did not accumulate significant amounts of the metabolic by-product acetate under these unlimited aerobic growth conditions.

Acetates↗
PubMed2006

Calibration of a 220Rn flow-through source.

Two reference methods have been established to calibrate the yield of a self-made 220Rn flow-through source. These reference methods include (a) a small scintillation cell flow-through method and (b) a gamma spectrometry method that takes advantage of an activated charcoal. The methods were validated theoretically and experimentally including intercomparison measurements. It is shown that the source is stable with a relative deviation less than 2.5% over 5 years, when the relative humidity ranged from 50 to 90%, and the temperature from 5 to 38 degrees C. The 220Rn monitor used in these reference methods is calibrated without a 220Rn gas source with determinate yield. On the basis of the calibration of the 220Rn source by the methods mentioned above, the accurate calibration of any 220Rn monitor by the self-made 220Rn source with stable yield is feasible.

Calibration↗
PubMed2007

A fermentation system designed to independently evaluate mixing and/or oxygen tension effects in microbial processes: development, application and performance.

In order to evaluate the independent effects of hydrodynamic conditions and/or oxygen tension on culture physiology and productivity, a fermentation system designed to control dissolved oxygen at constant power drawn (P/V) was developed. The system included a fully instrumented 14 l bioreactor coupled to a PC for data acquisition and control. Power drawn was measured (using a commercial torquemeter coupled to the shaft) and maintained constant by varying the agitation speed; while gas blending was used to control dissolved oxygen concentration. To validate the system, rheological-complex fermentations involving xanthan gum production and filamentous fungal cultivation (using Xanthomonas campestris and Trichoderma harzianum) were developed. In both cases, and despite the changing environmental conditions (due to increased broth viscosities and microbial respiration), both variables were controlled at the desired set points. Through such a system, a rigorous evaluation of the hydrodynamic conditions and/or oxygen tension on culture physiology and productivity is now feasible.

Bioreactors↗
PubMed2005

A continuous-flow perfusion system for the maintenance and NMR study of small tissue samples in vitro.

To describe and evaluate a novel perfusion system developed to maintain excised tissue in a flowing, oxygenated bathing solution during acquisition of nuclear magnetic resonance (NMR) data, and in addition allow precise data to be acquired continuously while altering the composition of the bathing solution surrounding the tissue. A chamber to house the tissue sample was constructed of interlocking sections of polyethylene tubing, and had approximate internal dimensions of 4 mm in diameter and 4 mm in height. Temperature-controlled, physiologically appropriate buffer solution was pumped via an infusion pump through the chamber, entering and exiting by way of small openings on either end. Immediately surrounding the polyethylene chamber was a tight-fitting four-loop solenoid RF coil. Measured proton NMR parameters were found to be fairly insensitive to the flow rate of the buffer if this coil was used only for reception and a larger-volume transmit-only coil was used for excitation. Temperature control of the sample was successfully implemented between 25 and 40 degrees C. The perfusion system was found to be resistant to the effects of flow rate, as well as a useful tool for the administration of drugs or agents to the tissue. Changes in buffer composition could be performed on the fly without the need to reposition the sample each time a change was made. This avoidance of repositioning was found to yield a fivefold improvement in the precision of T(2) spectral parameters (using frog sciatic nerve as a sample).

Animals↗
PubMed2005

Design and simulation of active biochip system.

To solve the problems existing in passive biochip systems, we designed a novel active biochip system. This system introduces negative pressure and controlling devices to adjust the antigen-antibody reaction on the nitrocellulose membrane. Computational simulation demonstrated that this system is a rapid, stable, robust and practical system that may enhance the efficiency of antigen-antibody reactions and improve the repeatability and accuracy of biochip analysis.

Antigen-Antibody Complex↗
PubMed2005

A system for micro/nano fluidic flow diagnostics.

A system for flow measurement in micro/nano fluidic components is presented. Microfabricated arrays of straight channels with noncircular cross-sections were used for flow rate measurement. The calculated flow rates in these channels were determined using a finite difference approximation method. A pneumatic pumping system was utilized to control the pressure drop across the channels and flow rates were measured by collecting the fluids on a sensitive balance. The experimental setup was validated using long narrow circular tubes that mimic the range of flow resistances characteristic of micro/nano fluidic devices. Two types of channels cross-section were investigated. The first type contained an array of channels that were approximately trapezoidal (microchannels, approximately 6.5 microm deep) in cross-section and exhibited flow rates of 27.7--119.4 micro L/min within a pressure range of 64.1--277.1 kPa (9.3--40.2 psi). The second type contained an array of channels that were approximately arc-shaped (nanochannels, approximately 600 nm deep) and generated flow rates of 0.29--0.99 micro L/min within a pressure range of 137.2--334.4 kPa (19.9--48.5 psi). The flow rates calculated by the finite difference approximation method were within 5.5% and 19.68% of the average experimental flow rates in the microchannels and nanochannels, respectively.

Computer Simulation↗
PubMed2005

Continuous-flow fractionation of animal cells in microfluidic device using aqueous two-phase extraction.

Monitoring of live cells is important in the field of medical science, diagnostics, biology, and the pharmaceutical industry. In this study, live and dead CHO-K1 (Chinese Hamster Ovary) cells were fractionated by continuous-flow extraction in a microfluidic device using immiscible aqueous two-phase extraction technique. The polymer solutions offered stable two-phase flows in microchannel without diffusive mixing. The fundamentals of aqueous two-phase extraction can support stable and reproducible recovery and separation of biomolecules in microfluidic devices. Polyethylene glycol 8000 (PEG 8000, 4%) and dextran T 500 (5%) were selected as model polymer solutions. The appropriate flow rates of polymer and cell solutions were suggested. The fractionation efficiency of live and dead CHO K-1 cells from the culture broth was compared in normal macroscale system and microfluidic device. The optimum pH for the fractionation was 6.6 in both the normal and micro-scale systems. The loss of target live cells by sedimentation was circumvented in microfluidic device because of the negligible effect of gravity on the sedimentation. Most live cells were distributed to PEG-rich phase, while dead cells were found at the interface of two polymer solutions in microchannel. In this case, the recovery and fractionation efficiency of live cells in the PDMS-based microfluidic device was 100% and 97.0%, respectively.

Animals↗
PubMed2005

Cross-talk problem on a fluorescence multi-channel microfluidic chip system.

Development of a compact fluorescence-based detection system for use in a micro-analytical system, such as a point-of-care diagnostic system, often requires a multi-channel microfluidic chip system. Since the materials used for microfluidic chips usually are transparent in the visible region and have a refractive indices higher than that of air or the surrounding environment, the fluorescence emission and scattered excitation light can propagate through the chip. We observed that such propagation can cause cross-talk between adjacent channels, and may become the major source of noise in the system and/or photo bleach the fluorescent samples in the adjacent channels, particularly for the small distances between the channels found in microfluidic chips, usually in order of several micro m. We monitored this cross-talk using fluorescein as a fluorescent sample and Mylar sheeting as a microfluidic chip material. We then discuss how this cross-talk can be avoided using a simple, inexpensive and effective method.

Artifacts↗
PubMed2005

Flat-chip microanalytical enzyme sensor for salivary amylase activity.

It is considered that measurement of salivary alpha-amylase activity is a useful tool for evaluating the sympathetic nervous system. The purpose of this research is to demonstrate a new design of a flat-chip microanalytical enzyme sensor (flat-chip sensor) for salivary amylase activity as a Micro-Electro-Mechanical Systems (MEMS), which may be used for wearable analytical systems. To meet this purpose, the biosensor needs to be miniaturized and to possess high-sensitivity. A pre-column and a flat-enzyme electrode were incorporated in a flow cell of volume 25.7 ml. In order to miniaturize the flow cell, two enzymatic membranes containing maltose phosphorylase obtained from Enterococcus hirae (MP membrane) and glucose oxidase and peroxidase (GOD-POD membrane) were immobilised on the same planar surface. As a result, a flat-chip sensor incorporating a flow cell as small as a C battery was produced. The optimum conditions of three parameters of the fabricated flat-chip sensor, the immobilising method of the enzymatic membrane, dropping volume of the mixed enzymatic solution and flow rate of the sample solution, were examined. An analytical system for 0-190 kU/l amylase activity with R(2) of 0.97 was fabricated with a sample volume of 50 microl. This research indicates the possibility of a wearable biosensor for continuous monitoring of salivary amylase activity.

Biosensing Techniques↗
PubMed2005

Self-actuated, thermo-responsive hydrogel valves for lab on a chip.

An easy to fabricate, thermally-actuated, self-regulated hydrogel valve for flow control in pneumatically driven, microfluidic systems is described. This microvalve takes advantage of the properties of the hydrogel, poly(N-isopropylacrylamide), as well as the aqueous fluid itself to realize flow control. The valve was designed for use in a diagnostic system fabricated with polycarbonate and aimed at the detection of pathogens in oral fluids at the location of the sample collection. The paper describes the construction and characterization of the hydrogel valves and their application for flow control, sample and reagent metering, sample distribution into multiple analysis paths, and the sealing of a polymerase chain reaction (PCR) reactor to suppress bubble formation. The hydrogel-based flow control is electronically addressable, does not require any moving parts, introduces minimal dead volume, is leakage and contaminant free, and is biocompatible.

Acrylic Resins↗
PubMed2005

A SU-8/PDMS hybrid microfluidic device with integrated optical fibers for online monitoring of lactate.

A microfluidic device with integrated optical fibres was developed for online monitoring of lactate. The device consists of a SU-8 waveguide, microfluidic channels and grooves for the insertion of optic fibres. It was fabricated by one-step photolithography of SU-8 polymer resist. Different channel widths (50-300 microm) were tested in terms of detection sensitivity. A wide range of flow rates were applied to investigate the influence of flow rate on signal fluctuations. The separation between optical fibre sensor and microfluidic channel and the width of fluidic channel have been optimized to maximize the detection sensitivity. It was revealed that 250 microm of channel width is the optimum light path length for a compromise between detection sensitivity and interference of ambient light. The independence of detection signals on flow rates was demonstrated within the range of flow rate (0.5-5 ml/hr) tested. Compared with conventional lactate detection, the device is proved to have high accuracy, relatively low limit of detection (50 mg/L) and reasonably fast response time (100 sec). The fabrication of device is simple and low cost. The present work has provided some fundamental data for further system optimization to meet specific detection requirements.

Biosensing Techniques↗
PubMed2005

Rapid mixing using two-phase hydraulic focusing in microchannels.

Rapid mixing is important in biomedical analysis. In this study, rapid mixing is obtained through two-phase hydraulic focusing in microchannels. Two mixing streams are focused by two sheath streams. Assuming a laminar flow in the channel, the spreading behavior of the two immiscible fluids is modeled and solved analytically. The results show that both viscosity ratio and flow rate ratio between the sheath flow and the sample flow can affect the focusing ratio. Thus, the mixing path of the sample flows can be adjusted by either viscosity ratio or flow rate ratio. Furthermore, an analytical model was proposed and solved for convective/diffusive mixing between the sample streams. According to this model, the focusing ratio is a key parameter for rapid mixing. A fully polymeric micro mixer was fabricated and tested for verification of the presented analytical models. The micromixer was fabricated by laser micromachining and adhesive bonding. The characterization results show the promising potential of mixing in microscale using two-phase hydraulic focusing.

Complex Mixtures↗
PubMed2006

Elucidating in vitro cell-cell interaction using a microfluidic coculture system.

This work presents a novel microfluidic coculture system that improves the accuracy of evaluating the interaction between cocultured cell types. A microfluidic coculture chip, fabricated by CO(2) laser direct-writing on polymethyl methacrylate (PMMA), was designed to separate two cell types using a microchannel, while permitting transfer of cellular media. The system has two up-stream wells and five down-stream wells. As an example, released inflammatory cytokines (e.g., interleukin-1 beta (IL-1 beta) and tumor necrosis factor-alpha (TNF-alpha)), activated in up-stream macrophages, flow through a microfluidic mixing system, generating linear concentration gradients in down-stream wells and inducing down-stream osteoblasts to release prostaglandin E2 (PGE2), a well-known bone resorption marker. Osteoblast viability was assessed by 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide (MTT) assay. This novel coculture system can be applied to evaluate cell-cell interaction while physically separating interacting cells.

Cell Communication↗
PubMed2006

Magnetic-based microfluidic platform for biomolecular separation.

A novel microfluidic platform for manipulation of micro/nano magnetic particles was designed, fabricated and tested for applications dealing with biomolecular separation. Recently, magnetic immunomagnetic cell separation has attracted a noticeable attention due to the high selectivity of such separation methods. Strong magnetic field gradients can be developed along the entire wire, and the miniaturized size of these current-carrying conductors strongly enhances the magnetic field gradient and therefore produces large, tunable and localized magnetic forces that can be applied on magnetic particles and confine them in very small spots. Further increases in the values of the generated magnetic field gradients can be achieved by employing miniaturized ferromagnetic structures (pillars) which can be magnetized by an external magnetic field or by micro-coils on the same chip. In this study, we demonstrate magnetic beads trapping, concentration, transportation and sensing in a liquid sample under continuous flow by employing high magnetic field gradients generated by novel multi-functional magnetic micro-devices. Each individual magnetic micro-device consists of the following components: 1. Cu micro-coils array embedded in the silicon substrate with high aspect ratio conductors for efficient magnetic field generation 2. Magnetic pillar(s) made of the magnetic alloy NiCoP for magnetic field focusing and magnetic field gradient enhancement. Each pillar is magnetized by its corresponding coil 3. Integrated sensing coil for magnetic beads detection 4. Microfluidic chamber containing all the previous components. Magnetic fields of about 0.1 T and field gradients of around 300 T/cm have been achieved, which allowed to develop a magnetic force of 3 x 10(-9) N on a magnetic particle with radius of 1 mum. This force is large enough to trap/move this particle as the required force to affect such particles in a liquid sample is on the order of approximately pN. Trapping rates of up to 80% were achieved. Furthermore, different micro-coil designs were realized which allowed various movement modes and with different step-sizes. These results demonstrate that such devices incorporated within a microfluidic system can provide significantly improved spatial resolution and force magnitude for quick, efficient and highly selective magnetic trapping, separation and transportation, and as such they are an excellent solution for miniaturized mu-total analysis systems.

Biopolymers↗