Efficacy of transgene expression in porcine skin as a function of electrode choice

Gene electrotransfer is a non-viral technique using electroporation for gene transfection. The method is widely used in the preclinical setting and results from the first clinical study in tumours have been published. However, the preclinical studies, which form the basis for the clinical trials, have mainly been performed in rodents and the body of evidence on electrode choice and optimal pulsing conditions is limited.We therefore tested plate and needle electrodes in vivo in porcine skin, which resembles human skin in structure. The luciferase (pCMV-Luc) gene was injected intradermally and subsequently electroporated. Simultaneously, studies with gene electrotransfer to porcine skin using plasmids coding for green fluorescent protein (GFP) and betagalactosidase were performed.Interestingly, we found needle electrodes to be more efficient than plate electrodes (p < 0.001) and electric field calculations showed that penetration of the stratum corneum led to much more homogenous field distribution at the DNA injection site. Furthermore, we have optimised the electric pulse regimens for both plate and needle electrodes using a range of high voltage and low voltage pulse combinations.In conclusion, our data support that needle electrodes should be used in human clinical studies of gene electrotransfer to skin for improved expression.     

Pumpless dispensing of a droplet by breaking up a liquid bridge formed by electric induction

Dispensing uniform pico‐to‐nanoliter droplets has become one of essential components in various application fields from high‐throughput bio‐analysis to printing. In this study, a new method is suggested and demonstrated for dispensing a droplet on the top plate with an inverted geometry by using electric field. The process of dispensing droplets consists of two stages: (i) formation of liquid bridge by moving up the charged fluid mass using the electrostatic force between the charges on the fluid mass and the induced charges on the substrate and (ii) its break‐up by the motion of the top plate. Different from conventional electrohydrodynamic methods, electric induction enables the droplets to be dispensed on various surfaces including non‐conducting substrate. The use of capillarity with an inverted geometry removes the need of external pumps or elaborates control for constant flow feed. The droplet diameter has been characterized as a function of the nozzle‐to‐plate distance and the plate moving velocity. The robustness of the present method is shown in terms of nozzle length and applied voltage. Finally, its practical applicability is confirmed by rendering a 19 by 24 array of highly uniform droplets with only 1.8% size variation without use of any active feedback control.     

Gene delivery to cells on a miniaturized multiwell plate for high-throughput gene function analysis

The transfectional microarray is a promising tool to conduct high-throughput analysis of gene function. In this study, a miniaturized multiwell plate was prepared by applying a silicone rubber sheet with holes on a gold electrode. The combination of electroporation and a miniaturized multiwell plate successfully achieved spatially controlled gene delivery with absence of cross-contamination between genes. Furthermore, we found that gene delivery efficiency was not dependent on conditions such as plasmid loading, electric field strength, and pulse duration.     

Research and application of a new rapid heat cycle molding with electric heating and coolant cooling to improve the surface quality of large LCD TV panels

The usage of rapid heat cycle molding (RHCM) has gained increasing attention in overcoming the limits of conventional injection molding (CIM) and improving the surface quality and mechanical properties of molded plastic products. In RHCM, the vario‐thermal mold temperature control system is the key technique because it directly affects the molding cycle time and the final part quality. In this study, a new RHCM technology with electric heating and coolant cooling was studied in detail. Two different RHCM mold structures for a large LCD TV panel were proposed and designed. The numerical simulation method was used to analyze the thermal response of the mold cavity surface at the heating stage and the thermal response of the resin melt at the cooling stage. The heating/cooling efficiency of the proposed electric heating RHCM system was evaluated. The thermal expansion analysis of mold cavity was implemented and the fixation of the cavity in molds was also optimized. The results showed that the electric‐heating mold with a separate cooling plate can efficiently enhance the heating efficiency. The thermal expansion of the cavity surface can be reduced by increasing the alleviating‐gap between the cavity and the cavity‐retainer plate. Then, the service lifetime of the electric‐heating mold can be improved. A RHCM production line with electric heating for the large LCD TV panel was constructed. Both the simulation and test production results indicate that the proposed electric heating RHCM technique can realize high‐temperature injection molding without increasing the molding cycle time. The surface appearance of the LCD TV panels was dramatically improved and the surface marks that usually occur in CIM process were eliminated completely. Copyright © 2009 John Wiley & Sons, Ltd.     

Electrokinetic diffusioosmotic flow of Ostwald-de Waele fluids near a charged flat plate in the thin double layer limit

Electrokinetic diffusioosmotic flow of Ostwald-de Waele, or power-law, fluids near a large charged flat plate is theoretically investigated for very thin double layers. Solutions to the flow velocity both up-close and far from the flat plate as well as the effective viscosity are presented for general values of the flow behavior index. Results show that given a wall zeta potential, ζ, diffusivity difference parameter, β, and constant imposed solute concentration gradient, both the near and far field diffusioosmotic flow velocities obtained for the respective dilatant and pseudoplastic liquids considerably deviate from those obtained for Newtonian liquids as found in previous literature. This likely suggests that the electrokinetic diffusioosmosis and its complementary effect of diffusiophoresis depend sensitively not only on the ζ-β parametric pair, but also on the possible non-Newtonian characteristics of the electrolytic liquid phase of the system. The theory presented herein can also be readily modified to model or describe electrodiffusioosmosis in power-law fluids, which is likely found in flow situations where the fluid non-Newtonian response, imposed solute concentration gradient, and an additional externally applied electric current density (or electric field) are of equal importance.     

Hot embossing of electrophoresis microchannels in PMMA substrates using electric heating wires

A simple method based on electric heating wires has been developed for the rapid fabrication of poly(methyl methacrylate) (PMMA) electrophoresis microchips in ordinary laboratories without the need for microfabrication facilities. A piece of stretched electric heating wire placed across the length of a PMMA plate along its midline was sandwiched between two microscope slides under pressure. Subsequently, alternating current was allowed to pass through the wire to generate heat to emboss a separation microchannel on the PMMA separation channel plate at room temperature. The injection channel was fabricated using the same procedure on a PMMA sheet that was perpendicular to the separation channel. The complete microchip was obtained by bonding the separation channel plate to the injection channel sheet, sealing the channels inside. The electric heating wires used in this work not only generated heat; they also served as templates for embossing the microchannels. The prepared microfluidic microchips have been successfully employed in the electrophoresis separation and detection of ions in connection with contactless conductivity detection.     

Performance of a Halo Ion Trap Mass Analyzer with Exit Slits for Axial Ejection

The halo ion trap (IT) was modified to allow for axial ion ejection through slits machined in the ceramic electrode plates rather than ejecting ions radially to a center hole in the plates. This was done to preserve a more uniform electric field for ion analysis. An in-depth evaluation of the higher-order electric field components in the trap was also performed to improve resolution. The linear, cubic and quintic (5th order) electric field components for each electrode ring inside the IT were calculated using SIMION (SIMION version 8, Scientific Instrument Services, Ringoes, NJ, USA) simulations. The preferred electric fields with higher-order components were implemented experimentally by first calculating the potential on each electrode ring of the halo IT and then soldering appropriate capacitors between rings without changing the original trapping plate design. The performance of the halo IT was evaluated for 1% to 7% cubic electric field (A ₄/A ₂) component. A best resolution of 280 (m/Δm) for the 51-Da fragment ion of benzene was observed with 5% cubic electric field component. Confirming results were obtained using toluene, dichloromethane, and heptane as test analytes.     

Electrophoresis of a colloidal sphere in a spherical cavity with arbitrary zeta potential distributions

An analytical study is presented for the quasi-steady electrophoretic motion of a dielectric sphere situated at the center of a spherical cavity when the surface potentials are arbitrarily nonuniform. The applied electric field is constant, and the electric double layers adjacent to the solid surfaces are assumed to be much thinner than the particle radius and the gap width between the surfaces. The presence of the cavity wall causes three basic effects on the particle velocity: (1) the local electric field on the particle surface is enhanced or reduced by the wall; (2) the wall increases the viscous retardation of the moving particle; and (3) a circulating electroosmotic flow of the suspending fluid exists because of the interaction between the electric field and the charged wall. The Laplace and Stokes equations are solved analytically for the electric potential and velocity fields, respectively, in the fluid phase, and explicit formulas for the electrophoretic and angular velocities of the particle are obtained. To apply these formulas, one has to calculate only the monopole, dipole, and quadrupole moments of the zeta-potential distributions at the particle and cavity surfaces. It is found that the contribution from the electroosmotic flow developing from the interaction of the imposed electric field with the thin double layer adjacent to the cavity wall and the contribution from the wall-corrected electrophoretic driving force to the particle velocities can be superimposed as a result of the linearity of the problem.     

Multistage electrophoresis II: treatment of a kinetic separation as a pseudoequilibrium process

An electrophoresis device is described which separates cells, particles, proteins and other separands by collecting samples having decreasing electrophoretic mobility in a train of inverted cavities while an electric field is applied between the inverted cavities and a sample cuvette containing a mixture of cells, particles, proteins or other separands. A circular plate is provided for the inverted cavities, and this circular plate is rotated to collect fractions. The system utilizes an innovative purification method that combines free electrophoresis and multistage extraction in an instrument capable of separating living cells, particles, and proteins in useful quantities at high concentrations. Most multistage processes are based on equilibrium separations, but electrophoresis is a kinetic separation; therefore, a pseudoequilibrium paradigm was developed for use in optimizing separation parameters including number of stages and electrophoresis time per stage. This paradigm allows the application of McCabe-Thiele type analysis, and it was calculated, for example, that two separands differing by 20% in electrophoretic mobility can be purified to 95% purity with acceptable yield in about seven stages. Laboratory experiments demonstrated a 95% purification in four stages of a separand originally present at 4% when electrophoretic mobilities differed by 80%.     

Drop formation via breakup of a liquid bridge in an AC electric field

Experimental results are presented for the study of drop formation mechanism in a newly proposed electrohydrodynamic (EHD) method of drop generation in an AC electric field. In the method, a small drop is generated in two stages. A pendant drop is elongated with large oscillation by an electric force in the first stage. Then, it undergoes formation and breakup of a liquid bridge between the upper nozzle and the insulator-coated lower flat plate in the second stage. It is found that there exists a resonant frequency for maximum oscillation, which leads to an efficient drop formation in the latter stage. It is also found that breakup of liquid bridge is accelerated by the electrowetting tension acting on the drop perimeter contacting the insulator-coated flat plate. Thus the whole procedure of drop formation depends heavily on the frequency of AC field and the properties of the insulator such as hydrophilicity, thickness, and the dielectric constant. It is demonstrated that a wide range of drop size, from picoliter to nanoliter, can be obtained by controlling such key parameters without changing the nozzle diameter.     

A hybrid aligned nematic reflective liquid crystal display driven by a fringe-field

We have designed a novel reflective nematic liquid crystal (LC) cell driven by a fringe electric field, in which the LCs are hybrid aligned in the initial state. Due to the hybrid alignment of the LC, the effective retardation value of the cell is greatly reduced when viewed in the normal direction and such a cell retardation value of 0.28 µm equals a quarter wave plate viewed in the normal direction. This means that the new reflective device can have a large cell gap of greater than 3 µm, which is advantageous when manufacturing the cell and, in addition, the device shows excellent electro-optic characteristics.     

Effects of nonequilibrium on velocity and plate height in reactive capillary electrophoresis

Models for velocity and plate height for reactive CE are developed under the formalism of generalized nonequilibrium theory, as described by Giddings. The resultant equations are consistent with chromatographic theory and validated with an independent stochastic simulation. Moreover, unlike prior methods for CE, this model allows calculation of thermodynamic equilibrium constants and kinetic rate constants from a single, undistorted peak. The theoretical development shows that velocity is directly dependent on the equilibrium constant and is independent of the rate constant. On the other hand, plate height varies little with equilibrium constant and is inversely proportional to rate constant. The ability to evaluate equilibrium constants from velocity and rate constants from plate height is most greatly influenced by electric field strength and mobility difference. The accuracy in calculated equilibrium constants is limited by mobility difference; however, the accuracy in rate constants is limited by plate height and equilibrium constant.     

Electrophoresis of a charge-regulated sphere at an arbitrary position in a charged spherical cavity

The electrophoresis of a charge-regulated spherical particle at an arbitrary position in a charged spherical cavity is modeled under conditions of low surface potential (<25 mV) and weak applied electric field (<25 kV/m). The charged cavity allows us to simulate the effect of electroosmotic flow, and the charge-regulated nature of the particle permits us to model various types of surface. The problem studied previously is reanalyzed based on a more rigorous electric force formula. In particular, the influences of various types of charged conditions on the electrophoretic behavior of a particle and the roles of all the relevant forces acting on the particle are examined in detail. Several new results are found. For instance, the mobility of a particle has a local minimum as the thickness of a double layer varies, which is not seen in the cases where the surface of a particle is maintained at a constant potential and at a constant charge density.     

Stability analysis for multi-jets electrospinning process modified with a cylindrical electrode

Nanosize fibers were fabricated using an extra-cylindrical electrode connected with single and multiple nozzles of an electrospinning process to stabilize the initial spun jets. To predict stability of spun jets, an electric field concentration factor (EFCF), which could be defined as a degree of a convergence of jets to a spinning axis, was introduced. The proposed parameter EFCF is utilized for the comparison of the experimental results for single and multi-nozzles electrospinning process. To consider the mass productivity of the multi-nozzles electrospinning supported by an auxiliary electrode, the weight of nanofibers that were spun to a rectangular shape target plate during 40 min was measured. The result indicates that the modified electrospinning technique shows a possibility as a useful method for increasing the production rate of nanofiber manufacturing.     

Electric potential distributions around discrete charges in a dielectric membrane—electrolyte solution system

Within the framework of the linearized Debye-Hückel theory an exact solution of the problem of calculating the electric potential caused by discrete fixed charges located at arbitrary positions with respect to a dielectric membrane-solution interface is presented. It takes into account the existence of an electrolyte solution on both sides of the membrane. Asymmetric ionic conditions are allowed for. For some interesting typical cases of fixed charge locations and electrolyte ionic strengths electric potential distributions were calculated and discussed. It is shown that, if the fixed charges were at or in front of the membrane surface, the characteristic distance of the electric potential decay was comparable to the Debye-Hückel length. At the opposite membrane surface only very small electric potentials can be observed. If, however, the fixed charge was placed below the membrane surface the electric potential in lateral direction and towards the other membrane surface largely increased. This effect was very sensitive to the position of the fixed charge with respect to the membrane surface.     

Time resolved X-ray diffraction studies of electric field induced layer motion in a chevron geometry smectic A liquid crystal device

Small angle time-resolved X-ray diffraction was used to monitor the behaviour of the smectic layers during the electric field induced planar to homeotropic transition in a smectic A cell possessing a chevron layer geometry. The liquid crystal material used was S3, from Merck Ltd, and was sandwiched in a 15 mum parallel plate device. The main features of the transition are the cooperative rotation of layers and the creation of an asymmetric chevron structure during the early stages of switching. The time scale for the planar-to-homeotropic transition in the device is approximately 5 s, at a temperature of 3 C below the nematic-to-smectic A phase transition and for an applied electric field of 2 V mum -1 (rms).     

Electrospray-ionization driven by dielectric polarization

A non-conductive piezo ceramic plate has been used to induce an electric field to generate an electrospray as ionization method for mass spectrometric determination. This technique decreases the risk of undesired discharges, induced by high electric currents. The applicability of the technique is demonstrated and compared with a commercial electrospray for mass spectrometric determination of reserpine and myoglobin.     

电感耦合等离子体质谱(ICP-MS)法测定农产品地土壤中铅、镉、铬消解方法的改进

采用赶酸电热板消解农产品地土壤中的重金属,电感耦合等离子体质谱(ICP-MS)法同时测定土壤中的铅、镉、铬。研究了酸体系及酸用量、赶酸电热板的升温程序,确定了最佳消解条件。通过统计50个批次土壤样品中加入的质控样,做出质控图,结果表明:各元素测定值均落在中心附近、上下警告线之内,批次内平行样品各元素相对标准偏差均小于5%。方法克服了传统的电热板消解法的缺点,弥补了高压罐消解法和微波消解法的不足,方法快速、准确,适合于大批量样品的分析。     

Model of a small-radius potential for molecular systems

The problem of particle mobility in a field of an arbitrary number of force centers with a small action radius and in a homogeneous electric or magnetic field is considered. An exact solution of the problem within the framework of the first-order perturbations over the wave function was obtained. The results may be used for the calculation of the polarizability and diamagnetic susceptibility of many-atom molecules.     

Electrical double layer interactions for spherical charge regulating colloidal particles

A new way of linearising the Poisson-Boltzmann equation is presented which is capable of producing accurate answers for moderately high surface charge and surface potentials. Unlike the Debye-Hückel approximation, the linearisation is based on the fact that electric potential variation, rather than the magnitude of the potential itself, remains small between two charged plates at close separation. The method can be applied quite generally to any type of surface. The cases of constant surface charge, the constant surfaceppotential and charge regulating systems are considered as examples. The results for constant charge and potentials as high as 5 μC/cm² and 200 mV are within 7% of the exact answers for plate separations of one Debye length or less. In contrast, the Debye-Hückel approximation produces answers that are at least an order of magnitude too large, for the same problems. The application of the method to the problem of calculating the electrostatic forces between charge regulated spherical particles is also addressed in some detail.