A conductive multi‐catalyst system consisting of Fe3O4 magnetic nanoparticles (MNPs) and oxidative enzymes co‐entrapped in the pores of mesoporous carbon is developed as an efficient and robust electrochemical biosensing platform. The construction of the nanocomposite begins with the incorporation of MNPs by impregnating Fe(NO3)3 on a wall of mesoporous carbon followed by heat treatment under an Ar/H2 atmosphere, which results in the formation of magnetic mesoporous carbon (MMC). Glucose oxidase (GOx) is subsequently immobilized in the remaining pore spaces of the MMC by using glutaraldehyde crosslinking to prevent enzyme leaching from the matrix. H2O2 generated by the catalytic action of GOx in proportion to the amount of target glucose is subsequently reduced into H2O by the peroxidase mimetic activity of MNPs generating cathodic current, which can be detected through the conductive carbon matrix. To develop a robust and easy‐to‐use electrocatalytic biosensing platform, a carbon paste electrode is prepared by mechanically mixing the nanocomposite or MMCs and mineral oil. Using this strategy, H2O2 and several phenolic compounds are amperometrically determined employing MMCs as peroxidase mimetics, and target glucose was successfully detected over a wide range of 0.5 × 10−3 to 10 × 10−3 M, which covers the actual range of glucose concentration in human blood, with excellent storage stability of over two months at room temperature. Sensitivities of the biosensor (19 to 36 nA mM−1) are about 7–14 times higher than that of the biosensor using immobilized GOx in mesoporous carbon without MNPs under optimized condition. The biosensor is of considerable interest because of its potential for expansion to any oxidases, which will be beneficial for use in practical applications by replacing unstable organic peroxidase with immobilized MNPs in a conductive carbon matrix.
The operation of an electrochemical real-time PCR system, based on intercalative binding of methylene blue (MB) with dsDNA, has been demonstrated. PCR was performed on a fabricated electrode-patterned glass chip containing MB while recording the cathodic current peak by measuring the square wave voltammogram (SWV). The current peak signal was found to decrease with an increase in the PCR cycle number. This phenomenon was found to be mainly a consequence of the lower apparent diffusion rate of the MB-DNA complex (D(b) = 6.82 × 10(-6) cm(2) s(-1) with 612 bp dsDNA) as compared to that of free MB (D(f) = 5.06 × 10(-5) cm(2) s(-1)). Utilizing this signal changing mechanism, we successfully demonstrated the feasibility of an electrochemical real-time PCR system by accurately quantifying initial copy numbers of Chlamydia trachomatis DNA templates on a direct electrode chip. A standard calibration plot of the threshold cycle (C(t)) value versus the log of the input template quantity demonstrated reliable linearity and a good PCR efficiency (106%) that is comparable to that of a conventional TaqMan probe-based real time PCR. Finally, the system developed in this effort can be employed as a key technology for the achievement of point-of-care genetic diagnosis based on the electrochemical real-time PCR.
The inertial migration of a two-dimensional elastic capsule in a Poiseuille flow was studied over the Reynolds number range 1 ≤ Re ≤ 100. The lateral migration velocity, slip velocity, and the deformation and inclination angle of the capsule were investigated by varying the lateral position, Reynolds number, capsule-to-channel size ratio (λ), membrane stretching coefficient (ϕ), and membrane bending coefficient (γ). During the initial transient motion, the lateral migration velocity increased with increasing Re and λ, but decreased with increases in ϕ, γ, and the lateral distance from the wall. On the other hand, the deformation of the capsule increased and the inclination angle became smaller as Re, ϕ, γ, and the distance from the wall decreased. The initial behavior of the capsule was influenced by variation in the initial lateral position (y(0)), but the equilibrium position of the capsule was not affected by such variation. The balance between the wall effect and the shear gradient effect determined the equilibrium position. As Re increased, the equilibrium position initially shifted closer to the wall and then moved toward the channel center. A peak in the equilibrium position was observed near Re = 30 for λ = 0.1, and the peak shifted to higher Re as λ increased. Depending on the lateral migration velocity, the equilibrium position moved toward the centerline for larger λ, but moved toward the wall for larger ϕ and γ.
An improved version of the immersed boundary (IB) method for simulating an initially circular or elliptic flexible ring pinned at one point in a uniform flow has been developed. The boundary of the ring consists of a flexible filament with tension and bending stiffness. A penalty method derived from fluid compressibility was used to ensure the conservation of the internal volume of the flexible ring. At Re = 100, two different flapping modes were identified by varying the tension coefficient for a fixed bending stiffness, or by changing the bending coefficient for a fixed tension coefficient. The optimal tension and bending coefficients that minimize the drag force of the flexible ring were found. Visualization of the vorticity field showed that the two flapping modes correspond to different vortex shedding patterns. We observed the hysteresis property of the flexible ring, which exhibits bistable states over a range of flow velocities depending on the initial inclination angle, i.e. one is a stationary stable state and the other a self-sustained periodically flapping state. The Reynolds number range of the bistability region and the flapping amplitude were determined for various aspect ratios a/b. For a/b = 0.5, the hysteresis region arises at the highest Reynolds number and the flapping amplitude in the self-sustained flapping state is minimized. A new bistability phenomenon was observed: for certain aspect ratios, two periodically flapping states coexist with different amplitudes in a particular Reynolds number range, instead of the presence of a stationary stable state and a periodically flapping state.
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