Specific detection of oxytetracycline using DNA aptamer-immobilized interdigitated array electrode chip

An electrochemical sensing system for oxytetracycline (OTC) detection was developed using ssDNA aptamer immobilized on gold interdigitated array (IDA) electrode chip. A highly specific ssDNA aptamer that bind to OTC with high affinity was employed to discriminate other tetracyclines (TCs), such as doxycycline (DOX) and tetracycline (TET). The immobilized thiol-modified aptamer on gold electrode chip served as a biorecognition element for the target molecules and the electrochemical signals generated from interactions between the aptamers and the target molecules was evaluated by cyclic voltammetry (CV) and square wave voltammetry (SWV). The current decrease due to the interference of bound OTC, DOX or TET was analyzed with the electron flow produced by a redox reaction between ferro- and ferricyanide. The specificity of developed EC-biosensor for OTC was highly distinguishable from the structurally similar antibiotics (DOX and TET). The dynamic range was determined to be 1-100 nM of OTC concentration in semi-logarithmic coordinates.     

Determination of pharmaceutical dosage forms via diffusion layer titration at an interdigitated microelectrode array

This paper presents a method for the analysis of drugs in dosage form. It is based on galvanostatic generation of oxidation agent from a suitable precursor on one segment of interdigitated microelectrode array (IDA) and its consecutive amperometric detection on second segment. High collection efficiency of this process in comparison to rotational ring disc electrode (RRDE) is a unique feature of IDA system. The transfer of oxidation agent can be influenced by addition of species, which reacts with oxidant. This influence can be used for its determination. Evaluation of generator-collector current dependence (diffusion layer titration curve) reveals the value of generator current I(genE) of the end-point of titration. I(genE) is proportional to the bulk phase concentration of determined species. The method was applied to the analysis of pharmaceuticals Antabus (Disulfiram Alpharma NOR, tetraethylthiuram disulfide (TETD)), a popular drug for alcoholism treatment, and Celaskon (vitamin C, Léciva CZE, ascorbic acid (AA)). From model samples analysis rather low detection limits, 9x10(-7) mol dm(-3), respectively, 4x10(-6) mol dm(-3), were estimated which enables trace content analysis of the drugs. A small size of IDA sensor also makes it suitable for microanalytical improvement.     

The detection of formaldehyde in textiles using interdigitated microelectrode array diffusion layer titration with electrogenerated hypobromite

An interdigitated microelectrode array (IDA) was applied to the determination of formaldehyde released from textiles produced in industry. The proposed method is based on formaldehyde reaction with hypobromite which is formed in weakly basic media by control current electrooxidation of bromide on the generator segment of the IDA array. The unreacted hypobromite diffuses through the gap between individually polarisable IDA segments and it is amperometrically detected on the collector segment of the IDA. The efficiency of this nonconvective transfer process in the absence of formaldehyde was substantially higher (78%) in comparison with that when using the rotating ring disc electrode. The influence of the added formaldehyde on the transfer process can be utilised to develop a simple and sensitive analytical procedure for formaldehyde detection with a detection limit of 4×10⁻⁶ mol dm⁻³.     

Indirect voltammetric detection of fluoride ions in toothpaste on a comb-shaped interdigitated microelectrode array

A novel technique based on dynamic electrochemistry for the detection of fluoride ions was developed. It is based on its strong complexation with ferric ion. Formed fluoroferric complex is cathodically inactive at the potential of the reduction of free ferric aquo ion. The voltammetric and amperometric response of platinum comb-shaped interdigitated microelectrode array is decreased after fluoride addition. This decrease serves for the quantification of fluoride ions added to the solution. The detection limit of 4.5 × 10⁻⁵ mol dm⁻³ was achieved when one of the segments of interdigitated microelectrode array (IDA) was used as an indicating electrode. The detection limit is about one order of magnitude lower than in the case of conventional platinum macroelectrode. In comparison with ISE electrodes this method is faster and also avoiding large error resulting from the antilogarithmization of ISE Nerstian response. The method was applied to the analysis of toothpaste.     

Glucose sensor based on redox-cycling between selectively modified and unmodified combs of carbon interdigitated array nanoelectrodes

We present a novel electrochemical glucose sensor employing an interdigitated array (IDA) of 1:1 aspect ratio carbon nanoelectrodes for the electrochemical-enzymatic redox cycling of redox species (ferricyanide/ferrocyanide) between glucose oxidase (GOx) and the two comb-shaped nanoelectrodes of the IDA. The carbon nanoelectrodes were fabricated using a simple, cost-effective, reproducible microfabrication technology known as the carbon-microelectromechanical-systems (C-MEMS) process. One comb (comb 1) of the IDA was selectively modified with GOx via the electrochemical reduction of an aryl diazonium salt, while the other comb (comb 2) remained unmodified; this facilitates electrochemically more active surface of comb 2, resulting in sensitive glucose detection. Ferricyanide is reduced to ferrocyanide by the GOx in the presence of glucose, and ferrocyanide diffuses to both combs of the IDA where it is oxidized. The limited electrochemical current collection at the surface-modified comb 1 is counterbalanced by the efficient redox cycling between the enzyme sites at comb 1 and the bare carbon surface of comb 2. Reducing the electrode-to-electrode gap between the two combs (gap = 1.9 μm) increases the diffusion flux of redox species at comb 2 hence, enhanced the sensitivity and limit of detection of the glucose sensor by ∼2.3 and ∼295 times, respectively at comb 2 compared to comb 1. The developed IDA-based glucose sensor demonstrated good amperometric response to glucose, affording two linear ranges from 0.001 to 1 mM and from 1 to 10 mM, with limits of detection of 0.4 and 61 μM and sensitivities of 823.2 and 70.0 μA mM⁻¹ cm⁻², respectively.     

Electrical impedance spectroscopy for detection of bacterial cells in suspensions using interdigitated microelectrodes

In this study, we present a new, simple and rapid impedance method to detect bacterial cells by making use of the impedance properties of bacterial cell suspensions using interdigitated microelectrodes. It was found that bacterial cell suspensions in deionized (DI) water with different cell concentrations could generate different electrical impedance spectral responses, whereas cell suspensions in phosphate buffered saline (PBS) solution could not produce any significant differences in impedance spectra in response to different cell concentrations. In DI water suspensions, impedance at 1 kHz decreased with the increasing cell concentrations in the suspensions. The impedance of cell suspensions in DI water was discussed and found that it was resulted from the cell wall charges and the release of ions or other osmolytes from the cells. A linear relationship between the impedance and the logarithmic value of the cell concentration was found in the cell concentration range from 10⁶ to 10¹⁰ cfu/ml, which can be expressed by a regression equation of Z (kΩ) = −2.06 log C (cells/20 μl) + 5.23 with R² = 0.98. The detection limit was calculated to be 3.45 × 10⁶ cfu/ml, which is comparable with many label-free immunosensors for detection of pathogenic bacteria reported in the literature. To achieve the selectivity of this method, we also demonstrated the feasibility of integrating magnetic separation to this impedance method. This study has demonstrated that bacterial cell concentration can be inferred by measuring the impedance of cell suspensions in DI water. This new detection mechanism could be an alternative to current impedance methods that have been reported for the detection of bacterial cells, e.g. impedance microbiology and electrical/electrochemical impedance biosensors.     

Interdigitated array microelectrode based impedance immunosensor for detection of avian influenza virus H5N1

Continuous outbreaks of avian influenza (AI) in recent years with increasing threat to animals and human health have warranted the urgent need for rapid detection of pathogenic AI viruses. In this study, an impedance immunosensor based on an interdigitated array (IDA) microelectrode was developed as a new application for sensitive, specific and rapid detection of avian influenza virus H5N1. Polyclonal antibodies against AI virus H5N1 surface antigen HA (Hemagglutinin) were oriented on the gold microelectrode surface through protein A. Target H5N1 viruses were then captured by the immobilized antibody, resulting in a change in the impedance of the IDA microelectrode surface. Red blood cells (RBCs) were used as biolabels for further amplification of the binding reaction of the antibody-antigen (virus). The binding of target AI H5N1 onto the antibody-modified IDA microelectrode surface was further confirmed by atomic force microscopy. The impedance immunosensor could detect the target AI H5N1 virus at a titer higher than 10³ EID₅₀/ml (EID₅₀: 50% Egg Infective Dose) within 2 h. The response of the antibody-antigen (virus) interaction was shown to be virus titer-dependent, and a linear range for the titer of H5N1 virus was found between 10³ and 10⁷ EID₅₀/ml. Equivalent circuit analysis indicated that the electron transfer resistance of the redox probe [Fe(CN)₆]^3−/4− and the double layer capacitance were responsible for the impedance change due to the protein A modification, antibody immobilization, BSA (bovine serum albumin) blocking, H5N1 viruses binding and RBCs amplification. No significant interference was observed from non-target RNA viruses such as Newcastle disease virus and Infectious Bronchitis disease virus. (The H5N1 used in the study was inactivated virus.)     

On-line microfluidic sensor integrated with a micro array electrode and enzyme-modified pre-reactor for the real-time monitoring of blood catecholamine

We developed an on-line microfluidic sensing device with an interdigitated array (IDA) electrode and a micro pre-reactor for the real-time monitoring of blood catecholamine (CA) and succeeded in the highly sensitive detection of dopamine (DA) in the presence of L-ascorbic acid (AA). Our device exhibits the lowest detection limit (110 ± 10 pM (S/N=3)), of reported catecholamine sensors. The improvement in sensitivity results from the high redox cycling of DA and the increase in the mass transfer rate per unit time onto the IDA electrode achieved by the flow measurement. The pre-reactor was integrated upstream in the micro flow channel to eliminate AA. A large number of rectangular shaped micropillars, which were modified with ascorbate oxidase, were formed in the pre-reactor to increase the surface area. The flow was disturbed by the two dimensional micropillar arrangement. This structure enables us to increase the elimination efficiency for AA. As a result, we achieved both the continuous and highly selective detection of 1 nM DA with complete elimination of 10 μM AA in the sample solution without employing any selective membrane such as Nafion, whose use reduces sensitivity due to the low diffusion coefficient of DA inside the membrane.     

Fabrication of interdigitated high-performance zinc oxide nanowire modified electrodes for glucose sensing

Diabetes is a metabolic disease with a prolonged elevated level of glucose in the blood leads to long-term complications and increases the chances for cardiovascular diseases. The present study describes the fabrication of a ZnO nanowire (NW)-modified interdigitated electrode (IDE) to monitor the level of blood glucose. A silver IDE was generated by wet etching-assisted conventional lithography, with a gap between adjacent electrodes of 98.80 μm. The ZnO-based thin films and NWs were amended by sol–gel and hydrothermal routes. High-quality crystalline and c-axis orientated ZnO thin films were observed by XRD analyses. The ZnO thin film was annealed for 1, 3 and 5 h, yielding a good-quality crystallite with sizes of 50, 100 and 110 nm, and the band gaps were measured as 3.26, 3.20 and 3.17 eV, respectively. Furthermore, a flower-modeled NW was obtained with the lowest diameter of 21 nm. Our designed ZnO NW-modified IDE was shown to have a detection limit as low as 0.03 mg/dL (correlation coefficient = 0.98952) of glucose with a low response time of 3 s, perform better than commercial glucose meter, suitable to instantly monitor the glucose level of diabetes patients. This study demonstrated the high performance of NW-mediated IDEs for glucose sensing as alternative to current glucose sensors.     

Microfluidic based impedance biosensor for pathogens detection in food products

A MEMS‐based impedance biosensor was designed, fabricated, and tested to effectively detect the presence of bacterial cells including E. coli O157:H7 and Salmonella typhimurium in raw chicken products using detection region made of multiple interdigitated electrode arrays. A positive dielectrophoresis based focusing electrode was used in order to focus and concentrate the bacterial cells at the centerline of the fluidic microchannel and direct them toward the detection microchannel. The biosensor was fabricated using surface micromachining technology on a glass substrate. The results demonstrate that the device can detect Salmonella with concentrations as low as 10 cells/mL in less than 1 h. The device sensitivity was improved by the addition of the focusing electrodes, which increased the signal response by a factor between 6 and 18 times higher than without the use of the focusing electrodes. The biosensor is selective and can detect other types of pathogen by changing the type of the antibody immobilized on the detection electrodes. The device was able to differentiate live from dead bacteria.     

检测禽流感H5亚型病毒的阻抗型免疫研究

研制了一种可用于H5亚型禽流感病毒快速检测的阻抗型免疫传感器。通过蛋白A将H5N1表面抗原血凝素(HA)的单克隆抗体固定于金叉指阵列微电极表面,并与待测溶液中的目标抗原H5N1进行免疫反应。在[Fe(CN)6]3"/4"溶液中进行电化学阻抗谱扫描,表征电极的表面修饰及抗原捕获过程。当H5N1病毒浓度在21～26 HA unit/50μL范围时,其浓度的对数值与叉指阵列微电极的电子传递阻抗的变化值呈线性关系,相关系数为0.9885;检出限为20 HA unit/50μL,检测时间为1 h。此传感器特异性好,灵敏度高,可以重复使用,在病原微生物快速检测领域具有良好的应用前景。     

Aminoglycoside array for the high-throughput analysis of small molecule-RNA interactions

Aminoglycoside-based oligosaccharides were bound to microtiter plates using noncovalent display with hybridization confirmed by mass spectrometry. Recognition of the saccharides by a model of the prokaryotic 16S aminoacyl site of the ribosomal decoding region was then determined. Tobramycin binding constants that correlated with surface plasmon resonance values for the model were effectively measured and novel means for binding aminoglycoside-based members were discovered for application to future aminoglycoside libraries.     

Preparation and characterization of Prussian blue nanowire array and bioapplication for glucose biosensing

Prussian blue nanowire array (PBNWA) was prepared via electrochemical deposition with polycarbonate membrane template for effective modification of glassy carbon electrode. The PBNWA electrode thus obtained was demonstrated to have high-catalytic activity for the electrochemical reduction of hydrogen peroxide in neutral media. This enabled the PBNWA electrode to show rapid response to H₂O₂ at a low potential of −0.1 V over a wide range of concentrations from 1 × 10⁻⁷ M to 5 × 10⁻² M with a high sensitivity of 183 μA mM⁻¹ cm⁻². Such a low-working potential also substantially improved the selectivity of the PBNWA electrode against most electroactive species such as ascorbic acid and uric acid in physiological media. A detection limit of 5 × 10⁻⁸ M was obtained using the PBNWA electrode for H₂O₂, which compared favorably with most electroanalysis procedures for H₂O₂. A biosensor toward glucose was then constructed with the PBNWA electrode as the basic electrode by crosslinking glucose oxidase (GOx). The glucose biosensor allowed rapid, selective and sensitive determination of glucose at −0.1 V. The amperometric response exhibited a linear correlation to glucose concentration through an expanded range from 2 × 10⁻⁶ M to 1 × 10⁻² M, and the response time and detection limit were determined to be 3 s and 1 μM, respectively.     

Methodology for the selection of suitable sensors for incorporation into a gas sensor array

An investigation into suitable mathematical techniques which can be used to select sensor components for a gas sensor array is reported. Data from a tin dioxide Taguchi semiconductor sensor array were obtained individually for various organic solvents and analysed using multivariate techniques, including principal component analysis, cluster analysis and star symbol plots. It was shown that the array data produced a series of characteristic response patterns for the analytes. It was also found that analytes of similar chemical nature had similar response patterns, indicating a correlation between sensor interaction and the chemical functional groups of the analyte. The multivariate techniques used proved to be very useful in enabling a suitable selection of the components of the array to be made by identifying which of the components or sensors were acting independently.     

Static droplet array for the synthesis of nonspherical microparticles

We reported a manually operated static droplet array (SDA)-based device for the synthesis of nonspherical microparticles with different shapes. The improved SDA structure and reversible bonding between poly(dimethylsiloxane) (PDMS) were used in the device for the large-scale synthesis and rapid extraction of nonspherical microparticles. To understand the device physics, the effects of flow rate, SDA well size, and shape on droplet generation performances were explored. The results indicated that droplet generation in SDA structures was insensitive to the flow rate, and monodisperse droplets were generated by the SDA-based device through manually pushing the syringe. Finally, we integrated four kinds of SDA structures in one device and successfully realized the synthesis and extraction of nonspherical microparticles with different shapes and materials. Our SDA-based device offers numerous advantages, such as simple manual operation, low equipment cost, controllable microparticle shapes and sizes, and large-scale production. Thus, it holds the potential to be used as a flexible tool for the production of nonspherical microparticles.     

A hydrogel based fluorescent micro array used for the characterization of liquid analytes

A fluorimetric micro spot array using non-specific recognition function is described for the analysis of liquid samples. The array was composed of binary mixtures of various fluorescence dyes which were embedded in a hydrogel matrix. The interactions between the fluorescent dyes and their molecular surrounding inside the hydrogel, influence their fluorescence wave length and intensities. The array was used for the characterization of solvent mixtures. Developed fluorescence patterns of the complete array as well as the fluorescence intensity changes of single spots were analysed. It was proven, that specific analytical information can be gained using this non-specific recognition approach. The identification of some alcoholic beverages is described as an example of the application of this method when used for quality control purposes. Analogous to the appellation “electronic nose” and “electronic tongue” the described micro spot array acts as an “optochemical tongue”.     

Comparison of a pump-around, a diffusion-driven, and a shear-driven system for the hybridization of mouse lung and testis total RNA on microarrays

In the present study, we demonstrate the benefits of a shear-driven rotating microchamber system for the enhancement of microarray hybridizations, by comparing the system with two commonly used hybridization techniques: purely diffusion-driven hybridization under coverslip and hybridization using a fully automated hybridization station, in which the sample is pumped in an oscillating manner. Starting from the same amount of DNA for the three different methods, a series of hybridization experiments using mouse lung and testis DNA is presented to demonstrate these benefits. The gain observed using the rotating microchamber is large: both in terms of analysis speed (up to tenfold increase) and in final spot intensity (up to sixfold increase). The gain is due to the combined effect of the hybridization chamber miniaturization (leading to a sample concentration increase if comparing iso-mass conditions) and the transport enhancement originating from the rotational shear-driven flow induced by the rotation of the chamber bottom wall.     

Capillary array electrophoresis for the research of asymmetric transformation reaction of L-proline to D-proline

One asymmetric transformation reaction of L-proline (L-Pro) to D-proline was studied by a home-made capillary array electrophoresis (CAE) for the first time. The aldehyde catalysts and the organic acid solvents for the asymmetric transformation reaction were rapidly screened and the enantiomeric excess values of the asymmetric product of L-Pro were directly obtained from the electrophoretogram of CAE.     

Quartz crystal microbalance sensor array for the detection of volatile organic compounds

A sensor array system consisting of five quartz crystal microbalance (QCM) sensors (four for measuring and one for reference) and an artificial neural network (ANN) method is presented for on-line detection of volatile organic compounds. Three ionic liquids, 1-butyl-3-methylimidazolium chloride (C₄mimCl), 1-butyl-3-methylimidazolium hexafluorophosphate (C₄mimPF₆), 1-dedocyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide (C₄mimNTf₂), and silicone oil II, which is widely used as gas chromatographic stationary phase, have been selected as sensitive coatings on the quartz surface allowing the sensor array effective to identify chemical vapors, such as toluene, ethanol, acetone and dichloromethane. The success rate for the qualitative recognition reached 100%. Quantitative analysis has also been investigated, within the concentration range of 0.6-6.1 mg/L for toluene, 0.9-7.5 mg/L for ethanol, 2.8-117 mg/L for dichloromethane, and 0.7-38 mg/L for acetone, with a prediction error lower than 8%.     

离子选择电极阵列土壤中硝态氮测定

土壤硝态氮反映土壤短期氮素供应水平,实时了解土壤硝态氮的含量为精准农业和农业面源污染防控提供支撑,因此,在线实时检测土壤硝态氮方法突破就显得十分迫切。土壤硝态氮中的硝酸根离子在土壤中的高水溶性和流动性为全固态硝酸根离子选择电极高敏感检测土壤中硝态氮提供了条件,固态硝态氮离子选择电极的离子选择膜反应硝酸根离子在被测溶液中的浓度。采用全固态硝酸根离子选择电极,且与温度电极和pH电极融合组成电极阵列对土壤饱和溶液中的硝态根离子进行检测。设计了高输入阻抗运算放大电路对电极信号进行采集,并通过微处理控制蠕动泵完成土壤硝态氮待测溶液连续流动测量及实时传输结果。实验结果表明,电极响应时间≤15 s,斜率-51.63 mV/decade,线性范围10-5-10-2.2 mol/L,最低检测限10-5.23 mol/L。相对标准差在0.78%-4.47%范围内,加标回收率均在90%-110%以内。与国家标准紫外可见分光光度法测试结果相比,相关系数（R2）为0.9952,为土壤硝态氮在现场检测奠定技术基础。