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Separation Channel (separation + channel)
Selected AbstractsFabrication of enclosed SU-8 tips for electrospray ionization-mass spectrometryELECTROPHORESIS, Issue 24 2005Santeri Tuomikoski Dr. Abstract We describe a novel electrospray tip design for MS which is fabricated completely out of SU-8 photoepoxy. A three-layer SU-8 fabrication process provides fully enclosed channels and tips. The tip shape and alignment of all SU-8 layers is done lithographically and is therefore very accurate. Fabrication process enables easy integration of additional fluidic functions on the same chip. Separation channels can be made with exactly the same process. Fluidic inlets are made in SU-8 during the fabrication process and no drilling or other postprocessing is needed. Channels have been fabricated and tested in the size range of 10,,m×10,,m,50,,m×200,,m. Mass spectrometric performance of the tips has been demonstrated with both pressure-driven flow and EOF. SU-8 microtips have been shown to produce stable electrospray with EOF in a timescale of tens of minutes. With pressure driven flow stable spray is maintained for hours. Taylor cone was shown to be small in volume and well defined even with the largest channel cross section. The spray was also shown to be well directed with our tip design. [source] Pump-free and low-cost negative pressure sampling device for rapid sample loading in MCEELECTROPHORESIS, Issue 24 2009Hongmei Hu Abstract A pump-free and low-cost negative pressure sampling device for injecting well-defined non-biased sample plugs into the separation channel of MCE was developed. It was composed of a pipet bulb, a 3-way electromagnetic valve and a single voltage supply at constant voltage. A sub-atmospheric pressure was created by hand-pressing air out of the pipet bulb and retained in it by switching the 3-way electromagnetic valve at cutoff position. During the sample loading stage, the sub-atmospheric pressure in the pipet bulb was applied via a 3-way electromagnetic valve to the headspace of the sealed sample waste reservoir (SW). A pinched sample plug was formed at the channel intersection in less than 0.5,s. Once the 3-way electromagnetic valve was switched to connect SW to ambient atmosphere to release the vacuum in SW, electrophoresis separation was consequently activated under the electric potentials applied. Experimental results demonstrated the pump-free negative pressure sampling device worked well in a wide vacuum degree ranged from ,250 to ,30,mbar with a satisfactory analytical precision. The sample consumption for each cycle was calculated to be 51,12,nL under the sampling pressure. Theoretical deduction indicates that the volume of the pipet bulb can be further reduced to 1,mL, which is critical for minimizing the sampling device for MCE. [source] A microfabricated CE chip for DNA pre-concentration and separation utilizing a normally closed valveELECTROPHORESIS, Issue 18 2009Chen-Hua Kuo Abstract A simple, sequential DNA pre-concentration and separation method by using a micro-CE chip integrated with a normally closed valve, which is activated by pneumatic suction, has been developed. The CE chip is fabricated using PDMS. A surface treatment technique for coating a polymer bilayer with an anionic charge is applied to modify the surface of the microchannel. A normally closed valve with anionic surface charges forms a nanoscale channel that only allows the passage of electric current but traps the DNA samples so that they are pre-concentrated. After the pre-concentration step, a pneumatic suction force is applied to the normally closed valve. This presses down the valve membrane, which reconnects the channels. The DNA samples are then moved into a separation channel for further separation and analysis. Successful DNA pre-concentration and separation has been achieved. Fluorescent intensity at the pre-concentration area is increased by approximately 3570 times within 1.9,min of operation. The signals from the separation of ,X174 DNA/HaeIII markers are enhanced approximately 41 times after 100,s of pre-concentration time, as compared with the results using a traditional cross-shaped micro-CE chip. These results clearly demonstrate that successful DNA pre-concentration for signal enhancement and separation analysis can be performed by using this new micro-CE chip. [source] Microchip electrophoresis in low-temperature co-fired ceramics technology with contactless conductivity measurementELECTROPHORESIS, Issue 14 2009Georg Fercher Abstract In this paper a novel micromachined contactless conductivity CE device produced in low temperature co-fired ceramics (LTCC) is introduced. The application of LTCC multilayer technology provides a promising method for the contactless detection of conductive compounds because of its increased dielectric constant compared with glass or plastics. The capacitive coupling of the excitation signal into the microchannel across the LTCC substrate is improved, resulting in better detection sensitivity. Two silver electrodes located externally at opposite sides at the end of the separation channel act as detector. Impedance variations in the channel are measured without galvanic contact between electrodes and fluid. Inorganic ions are separated in less than 1,min with this novel ceramic device. The limit of detection is 10,,M for potassium. [source] Nanostructured copolymer gels for dsDNA separation by CEELECTROPHORESIS, Issue 23 2008Fen Wan Abstract Pluronics are triblock copolymers of poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) that are able to form many different ordered nanostructures at appropriate polymer concentrations and temperatures in selective solvents. These nanostructured "gels" showed desirable criteria when used as DNA separation media, especially in microchip electrophoresis, including dynamic coating and viscosity switching. A ternary system of F127 (E99P69E99)/TBE buffer/1-butanol was selected as a model system to test the sieving performance of different nanostructures in separating dsDNA by CE. The nanostructures and their lattice constants were determined by small-angle X-ray scattering. Viscosity measurements showed the sol,gel transition phenomena. In addition to the cubic structure, successful electrophoretic separation of dsDNA in 2-D hexagonally packed cylinders was achieved. Results showed that without further optimization, ,X174 DNA,Hae III digest was well separated within 15,min in a 7-cm separation channel, by using F127/TBE/1-butanol gel with a 2-D hexagonal structure. A mechanism for DNA separations by those gels with both hydrophilic and hydrophobic domains is discussed. [source] A low-leakage sample plug injection scheme for crossform microfluidic capillary electrophoresis devices incorporating a restricted cross-channel intersectionELECTROPHORESIS, Issue 15 2008Chin-Lung Chang Abstract This study develops a crossform CE microfluidic device in which a single-circular barrier or a double-circular barrier is introduced at the cross-channel intersection. Utilizing a conventional crossform injection scheme, it is shown that these barriers reduce sample leakage and deliver a compact sample band into the separation channel, thereby ensuring an enhanced detection performance. A series of numerical and experimental investigations are performed to investigate the effects of the barrier type and the barrier ratio on the flow streamlines within the microchannel and to clarify their respective effects on the sample leakage ratio and sample plug variance during the injection process. The results indicate that a single-circular barrier injector with a barrier ratio greater than 20% and a double-circular barrier injector with a barrier ratio greater than 40% minimize the sample leakage ratio and produce a compact sample plug. As a result, both injectors have an excellent potential for use in high-quality, high-throughput chemical analysis procedures and in many other applications throughout the micro-total analysis systems field. [source] Characterization of voltage degradation in dynamic field gradient focusingELECTROPHORESIS, Issue 5 2008Jeffrey M. Burke Abstract Dynamic field gradient focusing (DFGF) is an equilibrium gradient method that utilizes an electric field gradient to simultaneously separate and concentrate charged analytes based on their individual electrophoretic mobilities. This work describes the use of a 2-D nonlinear, numerical simulation to examine the impact of voltage loss from the electrodes to the separation channel, termed voltage degradation, and distortions in the electric field on the performance of DFGF. One of the design parameters that has a large impact on the degree of voltage degradation is the placement of the electrodes in relation to the separation channel. The simulation shows that a distance of about 3,mm from the electrodes to the separation channel gives the electric field profile with least amount of voltage degradation. The simulation was also used to describe the elution of focused protein peaks. The simulation shows that elution under constant electric field gradient gives better performance than elution through shallowing of the electric field. Qualitative agreement between the numerical simulation and experimental results is shown. The simulation also illustrates that the presence of a defocusing region at the cathodic end of the separation channel causes peak dispersion during elution. The numerical model is then used to design a system that does not suffer from a defocusing region. Peaks eluted under this design experienced no band broadening in our simulations. Preliminary experimental results using the redesigned chamber are shown. [source] On-column conductivity detection in capillary-chip electrophoresisELECTROPHORESIS, Issue 24 2007Zhi-Yong Wu Professor Abstract On-column conductivity detection in capillary-chip electrophoresis was achieved by actively coupling the high electric field with two sensing electrodes connected to the main capillary channel through two side detection channels. The principle of this concept was demonstrated by using a glass chip with a separation channel incorporating two double-Ts. One double-T was used for sample introduction, and the other for detection. The two electrophoresis electrodes apply the high voltage and provide the current, and the two sensing electrodes connected to the separation channel through the second double-T and probe a potential difference. This potential difference is directly related to the local resistance or the conductivity of the solution defined by the two side channels on the main separation channel. A detection limit of 15,,M (600,ppb or 900,fg) was achieved for potassium ion in a 2,mM Tris-HCl buffer (pH,8.7) with a linear range of 2 orders of magnitude without any stacking. The proposed detection method avoids integrating the sensing electrodes directly within the separation channel and prevents any direct contact of the electrodes with the sample. The baseline signal can also be used for online monitoring of the electric field strength and electroosmosis mobility characterization in the separation channel. [source] Integration of continuous-flow sampling with microchip electrophoresis using poly(dimethylsiloxane)-based valves in a reversibly sealed deviceELECTROPHORESIS, Issue 14 2007Michelle W. Li Abstract Here we describe a reversibly sealed microchip device that incorporates poly(dimethylsiloxane) (PDMS)-based valves for the rapid injection of analytes from a continuously flowing stream into a channel network for analysis with microchip electrophoresis. The microchip was reversibly sealed to a PDMS-coated glass substrate and microbore tubing was used for the introduction of gas and fluids to the microchip device. Two pneumatic valves were incorporated into the design and actuated on the order of hundreds of milliseconds, allowing analyte from a continuously flowing sampling stream to be injected into an electrophoresis separation channel. The device was characterized in terms of the valve actuation time and pushback voltage. It was also found that the addition of sodium dodecyl sulfate (SDS) to the buffer system greatly increased the reproducibility of the injection scheme and enabled the analysis of amino acids derivatized with naphthalene-2,3-dicarboxaldehyde/cyanide. Results from continuous injections of a 0.39,nL fluorescein plug into the optimized system showed that the injection process was reproducible (RSD of 0.7%, n,=,10). Studies also showed that the device was capable of monitoring off-chip changes in concentration with a device lag time of 90,s. Finally, the ability of the device to rapidly monitor on-chip concentration changes was demonstrated by continually sampling from an analyte plug that was derivatized upstream from the electrophoresis/continuous flow interface. A reversibly sealed device of this type will be useful for the continuous monitoring and analysis of processes that occur either off-chip (such as microdialysis sampling) or on-chip from other integrated functions. [source] Poly(methylmethacrylate) and Topas capillary electrophoresis microchip performance with electrochemical detectionELECTROPHORESIS, Issue 16 2005Mario Castańo-Álvarez Abstract A capillary electrophoresis (CE) microchip made of a new and promising polymeric material: Topas (thermoplastic olefin polymer of amorphous structure), a cyclic olefin copolymer with high chemical resistance, has been tested for the first time with analytical purposes, employing an electrochemical detection. A simple end-channel platinum amperometric detector has been designed, checked, and optimized in a poly-(methylmethacrylate) (PMMA) CE microchip. The end-channel design is based on a platinum wire manually aligned at the exit of the separation channel. This is a simple and durable detection in which the working electrode is not pretreated. H2O2 was employed as model analyte to study the performance of the PMMA microchip and the detector. Factors influencing migration and detection processes were examined and optimized. Separation of H2O2 and L -ascorbic acid (AsA) was developed in order to evaluate the efficiency of microchips using different buffer systems. This detection has been checked for the first time with a microchip made of Topas, obtaining a good linear relationship for mixtures of H2O2 and AsA in different buffers. [source] Microchip capillary electrophoresis with a cellulose-DNA-modified screen-printed electrode for the analysis of neurotransmittersELECTROPHORESIS, Issue 15 2005Muhammad Johirul Abstract A microfluidic chip based on capillary electrophoresis coupled with a cellulose-single-stranded DNA (cellulose-ssDNA) modified electrode was used for the simultaneous analysis of dopamine (DA), norepinephrine (NE), 3,4-dihydroxy- L -phenylalanine (L -DOPA), 3,4-dihydroxyphenylacetic acid (DOPAC), and ascorbic acid (AA). The modification of the electrode improved the electrophoretic analysis performance by lowering the detection potential and enhancing the signal-to-noise characteristic without surface poisoning of the electrode. The sensitivity of the modified electrode was about 12 times higher than those of the bare ones. The test compounds were separated using a 62,mm long separation channel at the separation field strength of +200,V/cm within 220,s in a 10,mM phosphate buffer (pH,7.4). The most favorable potential for the amperometric detection was 0.7,V (vs. Ag/AgCl). A reproducible response (relative standard deviation of 1.3, 1.3, 2.1, 3.1, 3.4% for DA, NE, L -DOPA, DOPAC, and AA, respectively, for n,=,9) for repetitive sample injections reflected the negligible electrode fouling at the cellulose-ssDNA modified electrode. Square-wave voltammetric analyses reflected the sensitivities of the modified electrode for DA, NE, L -DOPA, DOPAC, and AA which were 1.78, 0.82, 0.69, 2.45, and 1.23,nC/µM with detection limits of 0.032, 0.93, 1.13, 0.31, and 0.62,µM, respectively. The applicability of this microsystem to real sample analysis was demonstrated. [source] A polymeric master replication technology for mass fabrication of poly(dimethylsiloxane) microfluidic devicesELECTROPHORESIS, Issue 9 2005Hai-Fang Li Abstract A protocol of producing multiple polymeric masters from an original glass master mold has been developed, which enables the production of multiple poly(dimethylsiloxane) (PDMS)-based microfluidic devices in a low-cost and efficient manner. Standard wet-etching techniques were used to fabricate an original glass master with negative features, from which more than 50 polymethylmethacrylate (PMMA) positive replica masters were rapidly created using the thermal printing technique. The time to replicate each PMMA master was as short as 20 min. The PMMA replica masters have excellent structural features and could be used to cast PDMS devices for many times. An integration geometry designed for laser-induced fluorescence (LIF) detection, which contains normal deep microfluidic channels and a much deeper optical fiber channel, was successfully transferred into PDMS devices. The positive relief on seven PMMA replica masters is replicated with regard to the negative original glass master, with a depth average variation of 0.89% for 26 ,m deep microfluidic channels and 1.16% for the 90 ,m deep fiber channel. The imprinted positive relief in PMMA from master-to-master is reproducible with relative standard deviations (RSDs) of 1.06% for the maximum width and 0.46% for depth in terms of the separation channel. The PDMS devices fabricated from the PMMA replica masters were characterized and applied to the separation of a fluorescein isothiocyanate (FITC)-labeled epinephrine sample. [source] A microfabricated capillary electrophoresis chip with multiple buried optical fibers and microfocusing lens for multiwavelength detectionELECTROPHORESIS, Issue 6 2005Suz-Kai Hsiung Abstract We present a new microfluidic device utilizing multiwavelength detection for high-throughput capillary electrophoresis (CE). In general, different fluorescent dyes are only excited by light sources with appropriate wavelengths. When excited by an appropriate light source, a fluorescent dye emits specific fluorescence signals of a longer wavelength. This study designs and fabricates plastic micro-CE chips capable of performing multiple-wavelength fluorescence detection by means of multimode optic fiber pairs embedded downstream of the separation channel. For detection purposes, the fluorescence signals are enhanced by positioning microfocusing lens structures at the outlets of the excitation fibers and the inlets of the detection fibers, respectively. The proposed device is capable of detecting multiple samples labeled with different kinds of fluorescent dyes in the same channel in a single run. The experimental results demonstrate that various proteins, including bovine serum albumin and ,-casein, can be successfully injected and detected by coupling two light sources of different wavelengths to the two excitation optic fibers. Furthermore, the proposed device also provides the ability to measure the speed of the samples traveling in the microchannel. The developed multiwavelength micro-CE chip could have significant potential for the analysis of DNA and protein samples. [source] Microfluidic device for capillary electrochromatography-mass spectrometryELECTROPHORESIS, Issue 21 2003Iulia M. Lazar Abstract A novel microfabricated device that integrates a monolithic polymeric separation channel, an injector, and an interface for electrospray ionization-mass spectrometry detection (ESI-MS) was devised. Microfluidic propulsion was accomplished using electrically driven fluid flows. The methacrylate-based monolithic separation medium was prepared by photopolymerization and had a positively derivatized surface to ensure electroosmotic flow (EOF) generation for separation of analytes in a capillary electrochromatography (CEC) format. The injector operation was optimized to perform under conditions of nonuniform EOF within the microfluidic channels. The ESI interface allowed hours of stable operation at the flow rates generated by the monolithic column. The dimensions of one processing line were sufficiently small to enable the integration of 4,8 channel multiplexed structures on a single substrate. Standard protein digests were utilized to evaluate the performance of this microfluidic chip. Low- or sub-fmol amounts were injected and detected with this arrangement. [source] A microfabricated hybrid device for DNA sequencingELECTROPHORESIS, Issue 21 2003Shaorong LiuArticle first published online: 6 NOV 200 Abstract We have created a hybrid device of a microfabricated round-channel twin-T injector incorporated with a separation capillary in order to extend the straight separation distance for high speed and long readlength DNA sequencing. Semicircular grooves on glass wafers are obtained using a photomask with a narrow line-width and a standard isotropic photolithographic etching process. Round channels are made when two etched wafers are face-to-face aligned and bonded. A two-mask fabrication process has been developed to make channels of two different diameters. The twin-T injector is formed by the smaller channels whose diameter matches the bore of the separation capillary, and the "usual" separation channel, now called the connection channel, is formed by the larger ones whose diameter matches the outer diameter of the separation capillary. The separation capillary is inserted through the connection channel all the way to the twin-T injector to allow the capillary bore flush with the twin-T injector channels. The total dead-volume of the connection is estimated to be , 5 pL. To demonstrate the efficiency of this hybrid device, we have performed four-color DNA sequencing on it. Using a 200 ,m twin-T injector coupled with a separation capillary of 20 cm effective separation distance, we have obtained readlengths of 800 plus bases at an accuracy of 98.5% in 56 min, compared to about 650 bases in 100 min on a conventional 40 cm long capillary sequencing machine under similar conditions. At an increased separation field strength and using a diluted sieving matrix, the separation time has been reduced to 20 min with a readlength of 700 bases at 98.5% base-calling accuracy. [source] Determination of oxalate in beer by zone electrophoresis on a chip with conductivity detectionJOURNAL OF SEPARATION SCIENCE, JSS, Issue 8 2003Marián Masár Abstract The use of a poly(methylmethacrylate) capillary electrophoresis chip, provided with a high sample load capacity separation system (a 8500 nL separation channel combined with a 500 nL sample injection channel) and a pair of on-chip conductivity detectors, for zone electrophoresis (ZE) determination of oxalate in beer was studied. Hydrodynamic and electroosmotic flows of the solution in the separation compartment of the chip were suppressed and electrophoresis was a dominant transport process in the separations performed on the chip. A low pH of the carrier electrolyte (3.8), implemented by aspartic acid and bis-tris propane, provided an adequate selectivity in the separation of oxalate from anionic beer constituents and, at the same time, also a sufficient sensitivity in its conductivity detection. Under our working conditions, this anion could be detected at a 0.5 ,mol/L concentration also in samples containing chloride (a major anionic constituent of beer) at a 1800 higher concentration. Such a favorable analyte/matrix concentration ratio made possible accurate and reproducible [typically, 2,5% relative standard deviation (RSD) values of the peak areas of the analyte in dependence on its concentration in the sample] determination of oxalate in 500 nL volumes of 20,50-fold diluted beer samples. Short analysis times (about 200 s), minimum sample preparation, and reproducible migration times of this analyte (0.5,1.0% RSD values) were characteristic for ZE on the chip. [source] Continuous capillary electrophoresis with ,ow injection and its application for determination of Ephedrine and Pseudo-ephedrine in Chinese medicinal preparationsBIOMEDICAL CHROMATOGRAPHY, Issue 8 2004Zhongwei Pan Abstract This article presents a new, simple and rapid continuous separation method by combination of ,ow injection with capillary electrophoresis designed for the analysis of basic traditional Chinese medicines. The device was produced using commercial capillary and components readily available in analytical laboratory. In double-T con,guration, the designed horizontal separation channel was 25 µm i.d. × 146 mm length (an effective separation length of 93 mm) quartz capillary, with two vertical elicitation arms produced from 0.5 mm i.d. pump tubing. The capillary was embedded in a 40 × 20 × 3 mm organic glass base. Using the double-T con,guration, continuous introduction of a series of samples was achieved. More than 3.00 resolution for ephedrine and pseudo-ephedrine were obtained using 100 mm borate buffer (pH 9.80) within 8 min in 25 µm separation channel with an electrical ,eld strength of 137 V/cm (UV detection at 215 nm). The linear calibration range was 50,1500 µg/mL (ephedrine, r = 0.9982; pseudo-ephedrine, r = 0.9990) for both analytes. The limits of detection were 2.65 µg/mL for ephedrine and 2.92 µg/mL for pseudo-ephedrine. In this device, the contents of ephedrine and pseudo-ephedrine in ,ve Chinese medicinal preparations were determined with RSDs (n = 5) in range 1.16,4.51% and recoveries in range 90.4,114.6%. Copyright © 2004 John Wiley & Sons, Ltd. [source] Multiple-point electrochemical detection for a dual-channel hybrid PDMS-glass microchip electrophoresis deviceELECTROPHORESIS, Issue 19 2009Mario Castańo-Álvarez Abstract A new PDMS-based dual-channel MCE with multiple-point amperometric detection has been evaluated. Electrophoresis has been optimised in a single-channel device. Pretreatment with 0.1,M NaOH is very important for increasing and stabilising the EOF. The precision is adequate for a day's work in terms of both peak current and migration time. The RSD of the peak current for five successive signals was 1.9, 2.4 and 3.1% for dopamine, p- aminophenol and hydroquinone. RSD for the migration time was always less than 1.3%, which demonstrates the stability of the EOF and the possibility of running multiple experiments in the same microchip. The adequate inter-microchip precision as well as the rapid and simple manufacturing procedure indicates the disposable nature of the PDMS microchips. A dual-channel device with very simple multiple-point amperometric detection is proposed here. Elasticity of the PDMS allows removing the polymer slightly and aligning gold wires working electrodes. Injection can be performed from each of the sample reservoirs or from both simultaneously. The distance between the separation channels is critical for obtaining adequate signals as well as the introduction of a high-voltage electrode in the buffer reservoir. Simultaneous measurement of the same analytes in both channels is possible by applying the same potential. Moreover, since no cross-separation is produced, different analytes or samples can be simultaneously measured. [source] Sequencing of real-world samples using a microfabricated hybrid device having unconstrained straight separation channelsELECTROPHORESIS, Issue 21 2003Shaorong Liu Abstract We describe a microfabricated hybrid device that consists of a microfabricated chip containing multiple twin-T injectors attached to an array of capillaries that serve as the separation channels. A new fabrication process was employed to create two differently sized round channels in a chip. Twin-T injectors were formed by the smaller round channels that match the bore of the separation capillaries and separation capillaries were incorporated to the injectors through the larger round channels that match the outer diameter of the capillaries. This allows for a minimum dead volume and provides a robust chip/capillary interface. This hybrid design takes full advantage, such as sample stacking and purification and uniform signal intensity profile, of the unique chip injection scheme for DNA sequencing while employing long straight capillaries for the separations. In essence, the separation channel length is optimized for both speed and resolution since it is unconstrained by chip size. To demonstrate the reliability and practicality of this hybrid device, we sequenced over 1000 real-world samples from Human Chromosome 5 and Ciona intestinalis, prepared at Joint Genome Institute. We achieved average Phred20 read of 675 bases in about 70 min with a success rate of 91%. For the similar type of samples on MegaBACE 1000, the average Phred20 read is about 550,600 bases in 120 min separation time with a success rate of about 80,90%. [source] Parallel separations of oligonucleotides with optically gated sample introduction on multi-channel microchipsJOURNAL OF SEPARATION SCIENCE, JSS, Issue 1-2 2004Hongwei Xu Abstract With the release of the human genome sequence, there has been increasing attention given to other genetic analyses, including the detection of genetic variations and fast sequencing of multiple samples for pharmacogenomics studies. Rapid injections of samples in multiplexed separation channels by optically gated sample introduction are shown here for DNA separation. Serial separations of four amino acids are shown in less than four seconds on a microchip with four multiplexed channels. Five short oligonucleotides have also been rapidly separated in 2% LPA with four channels using this technique. In addition, multiple unique samples have been simultaneously separated and five-base resolution has been demonstrated. [source] | ||||||||||