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Biomedical subjects

Hong-Yuan Chen

Publications and source records attributed to Hong-Yuan Chen.

At least 19 recordsLinked to original sources

Glucose microfluidic biosensors based on immobilizing glucose oxidase in poly(dimethylsiloxane) electrophoretic microchips.

Here we reported a novel microfluidic biosensor with an on-column immobilized enzyme microreactor. The fabrication approach of this biosensor is simple and the enzyme microreactors with controlled sizes can be placed at any desired position on the microchip. Taking glucose oxidase (GOx) as an example, electroosmotic flow (EOF) as a driving force and amperometry as a detection method, the performance of biosensors were modulated by changing the length of enzyme reactor from 0.5 cm to 3 cm, and the linear ranges were changed from 0-8.0 mM to 0-30.0 mM with the detection limits from 42 microM to 6.5 microM. The enzyme reactor remained its 65% activity after 23 days storage. It also showed good anti-interference ability and was used to quantify glucose in human serum samples.

Biosensing Techniques↗

Modification of poly(dimethylsiloxane) microfluidic channels with silica nanoparticles based on layer-by-layer assembly technique.

A hydrophilic poly(dimethylsiloxane) (PDMS) microchip with stable electroosmotic flow (EOF) was prepared by a simple and reproducible coating procedure with silica nanoparticles. The microchannel wall of PDMS chip was coated with a layer of poly(diallyldimethylammonium chloride) (PDDA) and then collected silica nanoparticles. The assembly was followed by contact angle, charge-coupled device (CCD) imaging, electroosmotic flow (EOF) measurements and electrophoretic separation experiments. Contact angle measurements revealed the coated surface was hydrophilic; the water contact angle for coated chips was 64 degrees compared with a water contact angle for native PDMS chips of 113 degrees . CCD images indicated a substantially more hydrophilic microchannel than native PDMS. We carried out a comparison and concluded that the EOF values on the coated PDMS chip were close to those values on the glass chip above pH 7.0. The coated channel had an excellent stability and reproducibility, RSD of EOF values (n=6) on native and coated PDMS microchip was 1.58 and 0.57%, respectively. Separation of dopamine and epinephrine was performed on the coated chip generated 1.40 x 10(5), 1.39 x 10(5) theoretical plates/m compared with the native PDMS chip of 0.79 x 10(5), 0.88 x 10(5), high resolution of 1.7 was achieved with a channel of 3.60 cm length.

Dimethylpolysiloxanes↗

Hollow PbWO4 nanospindles via a facile sonochemical route.

Novel nanosized lead tungstate (PbWO4) hollow spindles were successfully synthesized, for the first time, via a Pluronic P123- (EO20PO70EO20-) assisted sonochemical process. The triblock copolymer acted as a structure-directing agent and played a key role in the formation of the hollow spindles. An in situ micelle templating mechanism has been proposed for the possible formation mechanism of the hollow nanostructure. The optical properties of the final products were investigated. It is exciting that the as-prepared PbWO4 hollow structure shows extraordinarily high room-temperature photoluminescence intensity compared to the solid structures.

Journal Article↗

Direct electron transfer and electrocatalysis of hemoglobin adsorbed on mesoporous carbon through layer-by-layer assembly.

Using chitosan as an effective linker between CMK-3 and glassy carbon electrode surface, {Hb/CMK-3}n multilayer film-modified electrodes were constructed through layer-by-layer assembly. The morphology of thus-formed {Hb/CMK-3}n film was characterized by scanning electron micrographs, and the interaction of hemoglobin (Hb) with CMK-3 was studied by UV-vis spectroscopy and electrochemical methods. Under optimal conditions, {Hb/CMK-3}6 film showed a couple of stable and well-defined redox peaks at about -377 and -296 mV in pH 7.0 buffers. Furthermore, the {Hb/CMK-3}6 film displayed excellent electrocatalysis to the reduction of both H2O2 and O2. Based on thus-formed film and its direct electron transfer behavior, a novel biosensor was presented for the determination of H2O2 ranging from 1.2 to 57 muM with the detection limit of 0.6microM at S/N=3. CMK-3 provided a desirable matrix for protein immobilization and biosensor preparation.

Adsorption↗

Sonochemical route for self-assembled V2O5 bundles with spindle-like morphology and their novel application in serum albumin sensing.

Novel self-assembled V2O5 bundles with highly ordered superstructures and spindle-like morphology were synthesized by a rapid high-yielding sonochemical method. The as-prepared samples were characterized by X-ray diffraction, a field-emission scanning electron microscope, and a transmission electron microscope. The spindle-like V2O5 bundles are composed of several tens of homogeneous nanowires with diameters of 30-50 nm and lengths of 3-7 microm. A sensitive resonance light scattering method for the detection of bovine serum albumin (BSA) based on the self-assembled V2O5 bundles was developed. The results of the polarized resonance light scattering demonstrated that the Cabannes factor for the V2O5 bundles-BSA aggregates was BSA concentration-dependent.

Journal Article↗

Novel coupling mechanism-based imaging approach to scanning electrochemical microscopy for probing the electric field distribution at the microchannel end.

A novel coupling mechanism-based imaging approach to scanning electrochemical microscopy (SECM) was used to image the distribution of electric field at the end channel of a poly(dimethylsiloxane) (PDMS) capillary electrophoresis (CE) microchip in the absence of redox species. The coupling imaging mechanism was systematically investigated and qualitatively illustrated. It was proved that the distribution of solution potentials within the scanning plane caused a different reduction rate of water at the tip electrode, which led to the variation in tip current. Within the scanning plane, the solution potentials measured in the central area of the microchannel were usually higher than those measured outside. The SECM images showed a strong dependence on tip potential, tip-to-channel distance, and separation potential. According to the Tafel equation, SECM images were converted to parameters that directly showed the distribution of solution potential. Change in the solution potential along the central axial line of the microchannel was also continuously sensed by allowing the tip to approach the microchannel in the presence of high voltage. Using dopamine as a model compound, the effect of solution potential on electrochemical detection was estimated by detecting separation parameters.

Dimethylpolysiloxanes↗

Antimony(III)-doped PbWO4 crystals with enhanced photoluminescence via a shape-controlled sonochemical route.

Sb(III)-doped lead tungstate single crystals with controlled shapes and enhanced green emission have been synthesized via a facile, Pluronic P123 (EO(20)PO(70)EO(20))-assisted, sonochemical method. The surfactant Pluronic P123 was found to play a crucial role in the morphology control of the final products. The growth process was investigated by carefully following time-dependent experiments, and the oriented attachment process accompanying Ostwald ripening was proposed for the possible formation mechanism. The optical properties, such as the Raman spectra and PL spectra, of doped PbWO(4) were studied. Room-temperature photoluminescence of doped PbWO(4) samples with different morphologies at desired Sb doping concentrations showed greatly enhanced luminescence intensity compared to the undoped PbWO(4).

Journal Article↗

Determination of morphine and codeine in urine using poly(dimethylsiloxane) microchip electrophoresis with electrochemical detection.

In this paper, a poly(dimethylsiloxane) (PDMS) microchip with electrochemical (EC) detection was developed for rapid separation and detection of morphine and codeine. It was found that morphine and codeine were well separated within 140 s in phosphate buffer solution (PBS) (pH 6.6, 40 mM)-beta-cyclodextrin (beta-CD) (20 mM)-acetonitrile (30%, v/v). The detection limit was 0.2 microM for morphine and 1 microM for codeine. The protocol was successfully applied to monitoring the amount of morphine and codeine in human urine. Compared with the conventional methods, the presented method had many advantages such as lower instrument cost, less reagent consumption and shorter analysis time.

Acetonitriles↗

The direct electron transfer of myoglobin based on the electron tunneling in proteins.

The electron tunneling of the protein-polypeptide interactions was observed in the study of direct electron transfer of the myoglobin (Mb) on the electrode surface. The Mb was selected as a redox active protein and gelatine was selected to couple with Mb to form an electron tunneling. The electrochemical results indicated the presence of the electron tunneling and the direct electron transfer. The circular dichroism spectra suggested that the beta-sheet chain of gelatine could interact with alpha-helical chain to form an electron tunneling to promote the protein direct electrochemistry. The SDS-PAGE results proved that the electron tunneling between Mb and gelatine was noncovalent hydrogen bonds. The immobilized Mb showed a couple of quasi-reversible redox peaks with a formal potential of -0.37V (vs SCE) in 0.1 M pH 7.0 PBS. The modified electrodes displayed a rapid amperometric response to the reduction of oxygen, H2O2, and nitrite.

Circular Dichroism↗

A simple method to fabricate a chitosan-gold nanoparticles film and its application in glucose biosensor.

A novel film of chitosan-gold nanoparticles is fabricated by a direct and facile electrochemical deposition method and its application in glucose biosensor is investigated. HAuCl(4) solution is mixed with chitosan and electrochemically reduced to gold nanoparticles, which can be stabilized by chitosan and electrodeposited onto glassy carbon electrode surfaces along with the electrodeposition of chitosan. Then a model enzyme, glucose oxidase (GOD) is immobilized onto the resulting film to construct a glucose biosensor through self-assembly. The resulting modified electrode surfaces are characterized with both AFM and cyclic voltammetry. Effects of chitosan and HAuCl(4) concentration in the mixture together with the deposition time and the applied voltage on the amperometric response of the biosensor are also investigated. The linear range of the glucose biosensor is from 5.0 x 10(-5) approximately 1.30 x 10(-3) M with a Michaelis-Menten constant of 3.5 mM and a detection limit of about 13 microM.

Biosensing Techniques↗

Rapid, large-scale synthesis and electrochemical behavior of faceted single-crystalline selenium nanotubes.

This article describes a rapid, solution-phase approach to the large-scale synthesis of faceted single-crystalline Se nanotubes, in the presence of cetyltrimethylammonium bromide. The products were characterized by scanning electron microscopy, high-resolution transmission electron microscopy, X-ray diffraction, X-ray photoelectron spectroscopy, laser Raman spectrography, differential scanning calorimetry analysis, and thermogravimetric analysis. The growth mechanism of the Se nanotubes was investigated by a series of experiments, and the rationality of the faceted morphology model for the Se nanotubes was demonstrated from the energetics and geometry. Furthermore, the electrochemical behavior of the Se nanotubes was studied by voltammetric techniques.

Calorimetry, Differential Scanning↗

Nanocrystalline diamond modified gold electrode for glucose biosensing.

Boron-doped diamond has drawn much attention in electrochemical sensors. However there are few reports on non-doped diamond because of its weak conductivity. Here, we reported a glucose biosensor based on electrochemical pretreatment of non-doped nanocrystalline diamond (N-NCD) modified gold electrode for the selective detection of glucose. N-NCD was coated on gold electrode and glucose oxidase (GOx) was immobilized onto the surfaces of N-NCD by forming amide linkages between enzyme amine residues and carboxylic acid groups on N-NCD. The anodic pretreatment of N-NCD modified electrode not only promoted the electron transfer rate in the N-NCD thin film, but also resulted in a dramatic improvement in the reduction of the dissolved oxygen. This performance could be used to detect glucose at negative potential through monitoring the current change of oxygen reduction. The biosensor effectively performs a selective electrochemical analysis of glucose in the presence of common interferents, such as ascorbic acid (AA), acetaminophen (AP) and uric acid (UA). A wide linear calibration range from 10 microM to 15 mM and a low detection limit of 5 microM were achieved for the detection of glucose.

Biosensing Techniques↗

Off-line form of the Michaelis-Menten equation for studying the reaction kinetics in a polymer microchip integrated with enzyme microreactor.

We firstly transformed the traditional Michaelis-Menten equation into an off-line form which can be used for evaluating the Michaelis-Menten constant after the enzymatic reaction. For experimental estimation of the kinetics of enzymatic reactions, we have developed a facile and effective method by integrating an enzyme microreactor into direct-printing polymer microchips. Strong nonspecific adsorption of proteins was utilized to effectively immobilize enzymes onto the microchannel wall, forming the integrated on-column enzyme microreactor in a microchip. The properties of the integrated enzyme microreactor were evaluated by using the enzymatic reaction of glucose oxidase (GOx) with its substrate glucose as a model system. The reaction product, hydrogen peroxide, was electrochemically (EC) analyzed using a Pt microelectrode. The data for enzyme kinetics using our off-line form of the Michaelis-Menten equation was obtained (K(m) = 2.64 mM), which is much smaller than that reported in solution (K(m) = 6.0 mM). Due to the hydrophobic property and the native mesoscopic structure of the poly(ethylene terephthalate) film, the immobilized enzyme in the microreactor shows good stability and bioactivity under the flowing conditions.

Enzyme Stability↗

Proteins modification of poly(dimethylsiloxane) microfluidic channels for the enhanced microchip electrophoresis.

This report described proteins modification of poly(dimethylsiloxane) (PDMS) microfluidic chip based on layer-by-layer (LBL) assembly technique for enhancing separation efficiency. Two kinds of protein-coated films were prepared. One was obtained by successively immobilizing the cationic polyelectrolyte (chitosan, Chit), gold nanoparticles (GNPs), and protein (albumin, Albu) to the PDMS microfluidic channels surface. The other was achieved by sequentially coating lysozyme (Lys) and Albu. Neurotransmitters (dopamine, DA; epinephrine, EP) and environmental pollutants (p-phenylenediamine, p-PDA; 4-aminophenol, 4-AP; hydroquinone, HQ) as two groups of separation models were studied to evaluate the effect of the functional PDMS microfluidic chips. The results clearly showed these analytes were efficiently separated within 140 s in a 3.7 cm long separation channel and successfully detected with in-channel amperometric detection mode. Experimental parameters in two protocols were optimized in detail. The detection limits of DA, EP, p-PDA, 4-AP, and HQ were 2.0, 4.7, 8.1, 12.3, and 14.8 microM (S/N=3) on the Chit-GNPs-Albu coated PDMS/PDMS microchip, and 1.2, 2.7, 7.2, 9.8, and 12.2 microM (S/N=3) on the Lys-Albu coated one, respectively. In addition, through modification, the more homogenous channel surface displayed higher reproducibility and better stability.

Buffers↗

Patterning microbeads inside poly(dimethylsiloxane) microfluidic channels and its application for immobilized microfluidic enzyme reactors.

We propose a convenient and reliable approach for immobilizing microbeads on poly(dimethylsiloxane) (PDMS) microchips. It is built upon a simple fabrication procedure of PDMS chip through directly printing the master with an office laser printer which was described in our previous work (J. Chromatogr. A 2005, 1089, 270-275). On the printed toners used as the positive relief of the master, microbeads were immobilized by a thermal treatment and then transferred to the surface of the microchip by direct molding of the prepolymer on the master. With this approach, the region-selective immobilization of microbeads and the fabrication of PDMS microchips can be accomplished at the same time. Then, using these microbeads as supports, further modification with enzyme was achieved. Surface characteristics of the microbeads-modified PDMS microchannels were investigated with scanning electron microscope, atomic force microscope, and inverse fluorescence microscope. The electrokinetic properties of the native PDMS and the modified PDMS chips were also compared. Based on this approach, an immobilized glucose oxidase (GOD) reactor was constructed and the reaction using glucose as substrate was studied. All these experiments aim to show that the proposed approach may have a good potential in the study of biochemistry and other related areas.

Catalysis↗

EOF measurement by detection of a sampling zone with end-channel amperometry in microchip CE.

A simple method for EOF measurement by detection of sampling zones with end-channel amperometry in microchip CE is developed. This method is based on the principle of the Kohlrausch regulating function (KRF). A dilute electroactive ionic species is added to the BGE as a continuously eluting electrophore which is used as a probe. When a BGE-like sample at a different concentration is injected, a peak of sampling zone appears and the migration time is related to EOF. In a microchip CE with hybrid PDMS/glass channel, a cathodic EOF of the hybrid glass/PDMS microchip was measured by end-channel amperometry; the effects of sample concentration and different probes on EOF rate were discussed. The present method was applied to monitor EOF rates in glass and in PDMS microchips. There was no significant difference between the values of EOF rates measured by the present method and the current-monitoring method. Detection of nonelectroactive analytes K(+), Na(+), and Li(+) can also be accomplished by the indirect amperometric method. Hence, the effective mobility of analyte can be accurately obtained.

Conductometry↗