Biosensor on the base of polypyrrole modified ultramicroelectrode arrays

A novel approach for the construction of a micro biosensor on the base of amperometric enzyme microelectrode arrays is described in this paper. The technique contains the electrochemical deposition of different enzymes in a conducting organic polymer, e.g. polypyrrole, on a transducer built up with the help of microfabrication technology. The electrochemical characteristics of these microelectrode arrays can be compared to conventional microelectrodes with the advantage of higher current outputs. For the first time different model enzymes like glucose oxidase, choline oxidase and lactate oxidase have been tested showing the principal possibility to construct a micro biosensor on the base of ultramicroelectrode arrays.     

Effects of electrode surface modification with chlorotoxin on patterning single glioma cells

A microchip patterned with arrays of single cancer cells can be an effective platform for the study of tumor biology, medical diagnostics, and drug screening. However, patterning and retaining viable single cancer cells on defined sites of the microarray can be challenging. In this study we used a tumor cell-specific peptide, chlorotoxin (CTX), to mediate glioma cell adhesion on arrays of gold microelectrodes and investigated the effects of three surface modification schemes for conjugation of CTX to the microelectrodes on single cell patterning, which include physical adsorption, covalent bonding mediated by N-hydroxysuccinimide (NHS), and covalent bonding via crosslinking succinimidyl iodoacetate and Traut's (SIA-Traut) reagents. The CTX immobilization to microelectrodes was confirmed by high-resolution X-ray photoelectron spectroscopy. Physically adsorbed CTX showed better support for cell adhesion and is more effective in confining adhered cells on the electrodes than covalently-bound CTX. Furthermore, cell adhesion and spreading on microelectrodes were quantified in real-time by impedance measurements, which revealed an impedance signal from physically adsorbed CTX electrodes four times greater than the signal from covalently-bound CTX electrodes.     

Characterization and fabrication of disposable screen printed microelectrodes

We report the fabrication of disposable and flexible screen printed microelectrodes which are characterised with microscopy and cyclic voltammetry. These new type of screen printed electrochemical platforms consist of micro-sized graphite typically with radii of 60 to 100 microns are defined by an inert dielectric. The advantage of this type of electrochemical sensing platform is that each microelectrode is disposable and cost effective and thus does not require extensive cleaning or electrode pre-treatment between measurements. Prior to measurements the screen printed microelectrode needs only to be calibrated with a suitable redox probe, as is typically the case with microelectrodes. We show proof of concept that the screen printed microelectrodes are advantageous for electro-analytical measurements with the example of determination of lead via cathodic stripping voltammetry. The use of graphite screen printed microelectrodes allows comparable detection limits to that obtained in the literature at insonated boron doped diamond electrodes, without the need for power ultrasound – which otherwise limits the widespread applicability and ease of measurement.     

Individually addressable recessed gold microelectrode arrays with monolayers of thio-cyclodextrin nanocavities

Four, individually addressable 30 microm diameter, e-beam deposited, gold microelectrodes recessed by 6 microm were suitably spaced on a single substrate to avoid diffusional overlap between each microelectrode. The single substrate device was functionalised with thiolated alpha-, beta-, and gamma-cyclodextrin nanocavities without spacer groups to ensure close proximity of the cavities to the electrode surface. The microelectrodes were assessed in two stages. The e-beam deposited micron sized electrodes were characterized using models for recessed and inlaid microdisk electrodes. Subsequently, each individually addressable, atomically flat, micro-patterned gold electrode with thiolated CD ensembles was treated as a nanoporous electrode assembly. Theoretical and experimental results were compared using cyclic voltammetry. Atomic force microscopy was also used to characterise the modified microelectrodes. Comparisons were made with thiolated CDs deposited on macroelectrodes. This is the first report of the behaviour of immobilized CD nanocavities ensembles on atomically flat gold microelectrodes.     

Dielectrophoretic deformation of breast cancer cells for lab on a chip applications

This paper presents the development and experimental analysis of a curved microelectrode platform for the DEP deformation of breast cancer cells (MDA‐MB‐231). The platform is composed of arrays of curved DEP microelectrodes which are patterned onto a glass slide and samples containing MDA‐MB‐231 cells are pipetted onto the platform's surface. Finite element method is utilised to characterise the electric field gradient and DEP field. The performance of the system is assessed with MDA‐MB‐231 cells in a low conductivity 1% DMEM suspending medium. We applied sinusoidal wave AC potential at peak to peak voltages of 2, 5, and 10 V_pp at both 10 kHz and 50 MHz. We observed cell blebbing and cell shrinkage and analyzed the percentage of shrinkage of the cells. The experiments demonstrated higher percentage of cell shrinkage when cells are exposed to higher frequency and peak to peak voltage electric field.     

Electroanalysis of Bromate,Iodate and Chlorate at Tungsten Oxide Modified Platinum Microelectrode Arrays

This article describes the direct detection of Bromate, but also of chlorate and iodate, using modified arrays of platinum ultramicroelectrodes. The detection limits for these ions are IO =0.76 μM; BrO =2.34 μM and ClO =133.2 μM. The formation of a suitable tungsten oxide layer and the optimum analytical conditions are described. These oxide layers are formed electrochemically from hydrogen peroxide‐tungstanate solutions in acid conditions. Although film stability may be an issue, particularly at pH>3, these modified arrays of microelectrodes can be used for analytical purposes. After the analysis, the modifying oxide is easily removed and the Pt microelectrode array recovers its original properties.     

Active microeletronic chip devices which utilize controlled electrophoretic fields for multiplex DNA hybridization and other genomic applications

Microelectronic DNA chip devices that contain planar arrays of microelectrodes have been developed for multiplex DNA hybridization and a variety of genomic research and DNA diagnostic applications. These devices are able to produce almost any desired electric field configuration on their surface. This ability to produce well-defined electric fields allows charged molecules (DNA, RNA, proteins, enzymes, antibodies, nanobeads, and even micron scale semiconductor devices) to be electrophoretically transported to or from any microlocation on the planar surface of the device. Of key importance to the device function is the permeation layer which overcoats the microelectrodes. The permeation layer is generally a porous hydrogel material that allows water molecules and small ions (Na+, CI-, etc.) to freely contact the microelectrode surface, but impedes the transport of the larger analytes (oligonucleotides, DNA, RNA, proteins, etc.). The permeation layer prevents the destruction of DNA at the active microelectrode surface, ameliorates the adverse effects of electrolysis products on the sensitive hybridization reactions, and serves as a porous support structure for attaching DNA probes and other molecules to the array. In order to maintain rapid transport of DNA molecules, facilitate hybridization, and work within constrained current and voltage ranges, low conductance buffers and various electronic pulsing scenarios have also been developed. These active microelectronic array devices allow electrophoretic fields to be used to carry out accelerated DNA hybridization reactions and to improve selectivity for single nucleotide polymorphism (SNP), short tandem repeat (STR), and point mutation analysis.     

Heat-transference of toner masks onto conductive substrates: A rapid and easy way to produce microelectrode ensembles

A novel and simple method for fabrication of microelectrode ensembles is reported. This procedure is based on the heat-transference of toner masks onto conductive substrates such as gold, platinum, and glassy carbon. The percentage of printed toner masks was controlled by using a suitable graphic software. The heat transference of 100% black toner mask onto a conductive substrate isolated electrically its whole surface, in such a way no electrochemical response was established using the modified substrate. However, when the same substrates were coated with 99% black toner mask, few naked micro-areas of the conductive material were identified. Such modified substrate presented typical electrochemical behavior of microelectrode ensembles, which evinced the presence of exposed micro-areas of substrate to the bulk solution. Different percentages of coverage were evaluated and the microelectrode ensembles were characterized by cyclic voltammetry with good agreement with the established theory. Stripping voltammetry of metals was also performed demonstrating the successful application of these new ensembles in accordance with characteristics normally presented by microelectrodes. The main advantages of this new procedure are the simplicity, low-cost of equipments (LaserJet printer and thermal press equipment), and the high speed of production of microelectrode arrays on different substrates.     

Print-and-peel fabrication of microelectrodes

We describe a facile and expedient approach for the fabrication of arrays of microelectrodes on smooth substrates. A sequence of print-and-peel procedures allowed for the microfabrication of capacitance microsensors using office equipment and relatively simple wet chemistry. Microfluidic assemblies with reversibly adhered elastomer components allowed for the transfer of patterns of metallic silver, deposited via Tollens' reaction, onto the substrate surfaces. Electroplating of the silver patterns produced an array of micrometer-thick copper electrodes. Capacitance sensors were assembled by placing nonlithographically fabricated flow chambers over the microelectrode arrays. Triangular-waveform current-voltage (I/V) measurements showed a linear correlation between the capacitance of the print-and-peel fabricated devices and the dielectric constant of the samples injected into their flow chambers.     

粉末微电极——Ⅱ. 完全不可逆电极反应体系

在完全不可逆电极反应体系(氧和亚硫酰氯还原)中研究了粉末微电极的行为。可以用微多孔电极模型解释测得的实验结果, 粉末微电极技术可以广泛用于研究各种粉末催化剂的电催化行为并可能制备高灵敏、响应快的微型电化学传感器。     

Ultrathin Cell‐Membrane‐Mimic Phosphorylcholine Polymer Film Coating Enables Large Improvements for In Vivo Electrochemical Detection

Resisting biomolecule adsorption onto the surface of brain‐implanted microelectrodes is a key issue for in vivo monitoring of neurochemicals. Herein, we demonstrate that an ultrathin cell‐membrane‐mimic film of ethylenedioxythiophene tailored with zwitterionic phosphorylcholine (EDOT‐PC) electropolymerized onto the surface of a carbon fiber microelectrode (CFE) not only resists protein adsorption but also maintains the sensitivity and time response for in vivo monitoring of dopamine (DA). As a consequence, the as‐prepared PEDOT‐PC/CFEs could be used as a new reliable platform for tracking DA in vivo and would help understand the physiological and pathological functions of DA.     

The electroanalytical detection of hydrazine: a comparison of the use of palladium nanoparticles supported on boron-doped diamond and palladium plated BDD microdisc array

We show that both a random distribution of palladium nanoparticles supported on a BDD electrode or a palladium plated BDD microelectrode array can each provide a sensing platform for the electrocatalytic detection of hydrazine. The palladium nanoparticle modified electrode displays a sensitivity and limit of detection of 60 mA mol(-1) L and 2.6 microM respectively while the array has a sensitivity of 8 mA mol(-1) L with a detection limit of 1.8 microM. The beneficial cost implications of using palladium nano- or micro-particles in sensors compared to a palladium macroelectrode are evident. Interestingly the array of the nanoparticles shows similar sensitivity and limit of detection to the microelectrode array which probably indicates that the random distribution of the former leads to 'clumps' of nanoparticles that effectively act as microelectrodes.     

Ceramic-based multisite microelectrode array for rapid choline measures in brain tissue

A ceramic-based multisite microelectrode array to measure choline in vivo in brain tissues is described. The microelectrodes were linear to 200 μM choline (R²=0.999±0.001) with a detection limit of approximately 0.4 μM (S/N of 3) in both single microelectrode and self-referencing amperometric recording modes. The 90% rise time of the sensor was 1.4 s, allowing for rapid measures of choline. Good selectivity (>300:1) was observed over interferents such as ascorbic acid, uric acid, and DOPAC in the single microelectrode mode. However, a self-referencing recording mode was needed to remove potassium-evoked dopamine signals in rat striatum. In vivo measurements of choline in the rat brain are presented.     

基于薄膜电极的细胞电融合芯片

构建了一种薄膜电极阵列结构的细胞电融合芯片, 通过多聚物微通道底/顶层凸齿状的微电极, 以及多聚物微通道侧壁上溅射形成的一层离散式金属薄膜电极, 共同形成离散式"三明治"微电极结构. 该微电极结构可在微通道内部形成与传统凸齿状电极相似的非均匀分布的梯度电场, 通过介电电泳效应进行细胞控制及排队. 利用多聚物在芯片上填充了传统凸齿状电极的凹陷区, 克服了细胞在凹陷区无法有效排队与融合的缺点. 在芯片上利用K562细胞开展了基于介电电泳效应的细胞排队实验及基于可逆性电穿孔效应的电融合实验, 结果表明该芯片能够较好地实现细胞排队及融合, 融合所需控制电压低至10 V左右. 细胞排队率达99%以上, 几乎无细胞在绝缘物填充区(传统凸齿电极芯片的凹陷区)滞留, 细胞两两排队高于60%, 细胞融合效率约为40%, 比传统的细胞电融合方法和凸齿电极芯片有较大提高.     

Hydrodynamic voltammetry at an interdigitated electrode array in a flow channel: Part I. Numerical simulation

This paper describes the numerical simulation of convective diffusion at an interdigitated electrode array, consisting of multiple pairs of microelectrodes held at alternating applied potentials on one wall of a flow channel. The downstream microelectrode of each pair detects species generated at the upstream microelectrode. Concentration profiles in the channel, amperometric response, and signal-to-noise ratios for the detector electrodes are calculated. The simple backward implicit finite difference (BIFD) simulation approach is applicable for a wide range of channel conditions. The upper number of electrode pairs treatable is limited only by computational time. The agreement of the simulation with previous results for a single pair of electrodes under comparable conditions is very good. Substantial improvements in signal-to-noise ratio are predicted for the multi-electrode interdigitated electrode array relative to a single generator-detector pair of equal overall area. Electrode dimensions are discussed for optimum signal/noise ratio. Relative enhancement increases significantly with the number of generator-detector pairs.     

Voltammetric Characterization of Micro‐ and Submicrometer‐Electrode Arrays of Conical Shape for Electroanalytical Use

Densely packed micro‐ and submicrometer electrode arrays of platinum and gold (the nominal number, N, of electrodes in each array varies between 225 and 3600) are fabricated by photolithographic technique and vapor deposition processes of metal films. The electrodes are conical‐shaped and only their apexes are exposed to the electrolytic solution. The electrode arrays are characterized electrochemically in Ru(NH₃)₆Cl₃ aqueous solutions by using cyclic voltammetry at low scan rates, to establish the number of electrochemically active electrodes (N_ac) in each array; the geometric characterization is performed by scanning electron microscopy. All the investigated arrays provide steady‐state voltammograms, indicating diffusionally independent behavior of each microelectrode. The number of microelectrodes that are active in the fabricated arrays depends on microelectrode density. In particular, for the arrays with N=3600 and N=225, the fraction of active sites is about 45% and 90%, respectively. The analytical performance of some of the Pt version of the arrays is tested in hydrogen peroxide solutions, allowing verifying that linear calibration plots over the concentration range (0.1–20 mM) are obtained. This dynamic range is larger than that typically recorded at smooth polycrystalline platinum electrodes (0.5–5 mM), and the better performance is attributed to both the higher aspect ratio of the cone geometry and the higher mass transport associated to each microelectrode of the array. Reproducibility (within 3.5%, r.s.d.) and long‐term stability (within 5%, r.s.d., after 8 h continuous use) of the electrode systems are satisfactory. A low detection limit, based on the signal to noise ratio equal to 3, of 0.05 mM is found, which is adequate for a rapid monitoring of H₂O₂ in real samples and industrial processes.     

Microfluidics embedded with microelectrodes for electrostimulation of neural stem cells proliferation

The regeneration of the injured nerve and recovery of its function have brought attention in the medical field. Electrical stimulation(ES) can enhance the cellular biological behavior and has been widely studied in the treatment of neurological diseases. Microfluidic technology can provide a cell culture platform with the well-controlled environment. Here a novel microfluidic/microelectrode composite microdevice was developed by embedding the microelectrodes to the microfluidic platform, in whic...     

Fabrication of carbon microelectrodes with a micromolding technique and their use in microchip-based flow analyses

In this paper, we report a new technique to pattern carbon microelectrodes for use in microfluidics. This technique, termed micromolding of carbon inks, uses poly(dimethylsiloxane)(PDMS) microchannels to define the size of the microelectrode. First, PDMS microchannels of the approximate dimensions desired for the microelectrode are made by soft lithography. The PDMS is then reversibly sealed to a substrate and the microchannels are filled with carbon ink. After a heating step the PDMS mold is removed, leaving a carbon microelectrode with a size slightly smaller than the original PDMS microchannel. The resulting microelectrode (27 microm wide and 6 microm in height) can be reversibly sealed to a PDMS-based flow channel. Fluorescence microscopy showed that no leakage occurred around the chip/electrode seal, even up to flow rates of 10 microL min(-1). The electrode was characterized by microchip-based flow injection analysis. Injections of catechol in Hank's Balanced Salt Solution (pH 7.4), showed a linear response from 2 mM to 10 microM (r(2)= 0.995), with a sensitivity of 56.5 pA microM(-1) and an estimated limit of detection of 2 microM (0.27 picomole, S/N=3). Reproducibility of the electrode response was shown by repeated injections (n= 10) of a 500 microM catechol solution, resulting in a RSD of 4.6%. Finally, selectivity was demonstrated by coating the microelectrode with Nafion, a perfluoronated cation exchange polymer. Dopamine exhibited a response at the modified microelectrode while ascorbic acid was rejected by the Nafion-coating. These electrodes provide inexpensive detectors for microfluidic applications while also being viable alternatives to use of other carbon microelectrode materials, such as carbon fibers. Furthermore, the manner in which the microelectrodes are produced will be of interest to researchers who do not have access to state of the art microfabrication facilities.