Electrochemical Sensor Array Research Articles

Optimization of an Electrochemical DNA Assay by Using a 48‐Electrode Array and Redox Amplification Studies by Means of Scanning Electrochemical Microscopy

Sensible DNA: An electrochemical DNA assay based on specific Salmonella spp. capture probes and enzyme labeling with alkaline phosphatase was optimized by using a 48-electrode microarray and scanning electrochemical microscopy (SECM). SECM was further used to evaluate potential amplification strategies due to redox cycling. Due to insufficient detection limits and selectivity, electrochemical DNA sensors are not yet used as everyday tools in diagnostics. Here, we present an electrochemical DNA assay that is based on specific Salmonella spp. capture probes. Our optimization strategies and the specific features of related electrochemical DNA sensor arrays, which are comprised of a chip with 48 gold electrodes, are also described. A ssDNA monolayer is formed by chemisorption of the thiol-modified capture strand on the different gold electrodes of the array after spotting with a needle spotter. The assay parameters were optimized for the use of minimum amounts of sample and reagents and short assay times. Scanning electrochemical microscopy (SECM) has been used to visualize the local activity of an enzyme label used for amplified hybridization detection at high lateral resolution. The potential of SECM to further amplify the sensor signal by means of redox cycling is demonstrated by using single-stranded DNA capture probe modified gold microelectrodes as SECM tips. The detection limit of the proposed DNA sensor is shown to be in the femtomolar range without redox cycling amplification.

Sensors as tools for quantitation, nanotoxicity and nanomonitoring assessment of engineered nanomaterials

The discovery of fullerenes in 1985 has ushered in an explosive growth in the applications of engineered nanomaterials and consumer products. Nanotechnology and engineered nanomaterials (ENMs) are being incorporated into a range of commercial products such as consumer electronics, cosmetics, imaging and sensors. Nanomaterials offer new possibilities for the development of novel sensing and monitoring technologies. Nanosensors can be classified under two main categories: (i) Nanotechnology-enabled sensors or sensors that are themselves nanoscale or have nanoscale materials or components, and (ii) Nanoproperty-quantifiable sensors or sensors that are used to measure nanoscale properties. The first category can eventually result in lower material cost, reduced weight and energy consumption. The second category can enhance our understanding of the potential toxic effects of emerging pollutants from nanomaterials including fullerenes, dendrimers, and carbon nanotubes. Despite the enormous literatures and reviews on Category I sensors, there are few sensors to measure nanoscale properties or sensors belonging to Category II. This class of nanosensors is an area of critical interest to nanotoxicology, detection and risk assessment, as well as for monitoring of environmental and/or biological exposure. This article discusses emerging fields of nanotoxicology and nanomonitoring including the challenges of characterizing engineered nanomaterials and the potentials of combining existing analytical techniques with conventional cytotoxicity methods. Two case studies are provided for development of Category II nanosensors for fullerene nanoparticles and quantum dots. One highlights the uniqueness of a portable, dissolved oxygen electrochemical sensor arrays capable of detecting the ENMs as well as provide rapid nanotoxicological information. This review has shown that addressing the complex and critical issues surrounding the environmental transformation and toxicity of ENMs must be accompanied by the creation of new approaches or further developments of existing instrumentation.

Ligase-based multiple DNA analysis by using an electrochemical sensor array

We herein report an electrochemical biosensor for the sequence-specific detection of DNA with high discrimination ability for single-nucleotide polymorphisms (SNPs). This DNA sensor was constructed by a pair of flanking probes that “sandwiched” the target. A 16-electrode electrochemical sensor array was employed, each having one individual DNA capture probe immobilized at gold electrodes via gold-thiol chemistry. By coupling with a biotin-tagged detection probe, we were able to detect multiple DNA targets with a single array. In order to realize SNP detection, a ligase-based approach was employed. In this method, both the capture probe and the detection probe were in tandem upon being hybridized with the target. Importantly, we employed a ligase that specifically could ligate tandem sequences only in the absence of mismatches. As a result, when both probes were complementary to the target, they were ligated in the presence of the ligase, thus being retained at the surface during the subsequent stringent washing steps. In contrast, if there existed 1-base mismatch, which could be efficiently recognized by the ligase, the detection probe was not ligated and subsequently washed away. A conjugate of avidin–horseradish peroxidase was then attached to the biotin label at the end of the detection probe via the biotin–avidin bridge. We then electrochemically interrogated the electrical current for the peroxidase-catalyzed reduction of hydrogen peroxide. We demonstrated that the electrochemical signal for the wild-type DNA was significantly larger than that for the sequence harboring the SNP.

Electrochemical sensor array forin situmeasurements of hydrogen peroxide concentration in high temperature water

AbstractA common index of corrosive conditions in boiling water reactors (BWRs) is the electrochemical corrosion potential (ECP), which is determined by a combination of factors such as specimen surface conditions and oxidant concentrations (e.g. O2, H2O2 and other corrosive radiolytic species). ECP sensors can be applied to determine corrosive conditions directly at elevated temperature, but a disadvantages of this approach is the broad plateau observed in the range 10–100 ppb H2O2. To determine the concentration [H2O2] directly in high temperature water, an array consisting of an ECP sensor and a complex impedance sensor has been developed. From the results, it is concluded: (1) the radii of the low frequency semicircle of the frequency dependent complex impedance (FDCI) data (Cole–Cole plots) decreased with increasing [H2O2]. The radii for the specimen exposed to 100 ppb H2O2 were determined by the electric resistance of the oxide film, and had reached the saturation level caused by saturation of oxide...

Integrated Electrochemical Sensor Array for On-Line Monitoring of Yeast Fermentations

This paper describes the design, modeling, and experimental characterization of an electrochemical sensor array for on-line monitoring of fermentor conditions in both miniaturized cell assays and in industrial scale fermentations. The viable biomass concentration is determined from impedance spectroscopy. As a miniaturized electrode configuration with high cell constant is applied, the spectral conductivity variation is monitored instead of the permittivity variation. The dissolved oxygen concentration is monitored amperometrically using an ultramicroelectrode array, which is shown to have negligible flow dependence. pH is monitored using an ion-sensitive field effect transistor (ISFET), and a platinum thermistor is included for temperature measurements. All sensors were shown to be sufficiently accurate within the range relevant to yeast fermentations. The sensor array is shown to be very stable and durable and withstands steam-sterilization.

Electrochemical Sensor Array for Glucose Monitoring Fabricated by Rapid Immobilization of Active Glucose Oxidase within Photochemically Polymerized Hydrogels

Currently, monitoring blood glucose levels for diabetic patients is invasive and painful, involving pricking the finger to obtain a blood sample three to four times daily. The need for frequent tests and pain involved with testing leads to poor compliance. In order to raise compliance, we propose to create an implantable electrochemical sensor array that would monitor glucose levels continuously. Glucose sensor arrays were fabricated on gold electrodes on flexible polyimide sheets by photopolymerization of the biocompatible polymer poly(ethylene glycol) diacrylate (PEG-DA) to develop hydrogels and encapsulate the sensing elements. Using conventional silicon fabrication methods, arrays of five gold microdisk electrodes were fabricated using lift-off photolithography and sputtering techniques. A redox polymer was then attached electrostatically to the electrode, and glucose oxidase was entrapped inside the hydrogel on the array of electrodes by ultraviolet-initiated photopolymerization of PEG-DA. When the array of fabricated sensors was sampled together the elements behaved like one large electrode with peak current equivalent to the sum of individual array elements. The enzyme, glucose oxidase, catalyzed the oxidation of glucose and then exchanged electrons with the redox polymer in the hydrogel. The entrapped glucose oxidase was found to respond linearly to increasing glucose concentrations (0-360 mg/dl), as determined using cyclic voltammetry. The fabricated microarray sensors were individually addressable and showed no cross talk between adjacent array elements as assessed using cyclic voltammetry. We have fabricated an array of glucose sensors on flexible polyimide sheets that exhibits the desired linear response in the biological range.

An Electrochemical Sensor Array for In-situ Measurements of Oxide Film Electric Resistance in High Temperature Water

Corrosive conditions in primary cooling systems of boiling water reactors are determined by oxidants, e.g., O2, H2O2 and other corrosive radiolytic species. A high temperature high pressure water loop, which controls H2O2 concentration with minimal O2 co-existence, was constructed in order to evaluate the effects of H2O2 on corrosive conditions, and then static and dynamic responses of stainless steel specimens exposed to H2O2 and O2 in high temperature pure water were evaluated by analyzing ECP (electrochemical corrosion potential) and FDCI (frequency dependent complex impedance). At the same time, in- situ measurements were carried out to determine changes in oxide film electric resistance. From the multilateral high temperature measurements, it was concluded that H2O2 concentrations could be determined by in- situ measurement based on a sensor complex for ECP and FDCI and that ECP and [H2O2] data could be used to evaluate occurrence and propagation of intergranular corrosion cracking of reactor internal components.

Electrochemical Sensor Array and Its Application to Real Time Imaging of a Brain Slice

An electrochemical sensing system using a planar microelectrode array has been developed to monitor biological molecules with relatively high special and temporal resolutions. This enables us a real time imaging of the biological molecules release from a tissue invasively. In this study, we have established a multichannel hydrogen peroxide (H2O2) sensing system to monitor the real time H2O2 distribution in a tissue using a planar sensor array. H2O2 has been recognized in association with the pathology of neurological diseases because it is a by-product of a degenerative reaction of reactive oxygen species, one of the major causes of oxidative stress in mammalian cells. The sensor array is based on a 64-channel ITO electrode array of 50x50 μm electrodes modified with an enzyme, horseradish peroxidase, and an electron transfer mediator. Then we place a cultured rat hippocampal slice on the array and measure the current at each sensor using a multipotentiostat. When we introduce bicuculline into the solution as a stimulant, in the presence of a catalase inhibitor, we can observe a distinct increase in the H2O2 concentration. This real-time H2O2 distribution monitoring system will be a powerful tool with which to explore the neuronal cell death mechanism in biological systems.

Use of Electrochemical DNA Biosensors for Rapid Molecular Identification of Uropathogens in Clinical Urine Specimens

We describe the first species-specific detection of bacterial pathogens in human clinical fluid samples using a microfabricated electrochemical sensor array. Each of the 16 sensors in the array consisted of three single-layer gold electrodes-working, reference, and auxiliary. Each of the working electrodes contained one representative from a library of capture probes, each specific for a clinically relevant bacterial urinary pathogen. The library included probes for Escherichia coli, Proteus mirabilis, Pseudomonas aeruginosa, Enterocococcus spp., and the Klebsiella-Enterobacter group. A bacterial 16S rRNA target derived from single-step bacterial lysis was hybridized both to the biotin-modified capture probe on the sensor surface and to a second, fluorescein-modified detector probe. Detection of the target-probe hybrids was achieved through binding of a horseradish peroxidase (HRP)-conjugated anti-fluorescein antibody to the detector probe. Amperometric measurement of the catalyzed HRP reaction was obtained at a fixed potential of -200 mV between the working and reference electrodes. Species-specific detection of as few as 2,600 uropathogenic bacteria in culture, inoculated urine, and clinical urine samples was achieved within 45 min from the beginning of sample processing. In a feasibility study of this amperometric detection system using blinded clinical urine specimens, the sensor array had 100% sensitivity for direct detection of gram-negative bacteria without nucleic acid purification or amplification. Identification was demonstrated for 98% of gram-negative bacteria for which species-specific probes were available. When combined with a microfluidics-based sample preparation module, the integrated system could serve as a point-of-care device for rapid diagnosis of urinary tract infections.

Study of Halitosis-Substance Sensing at Low Concentration Using an Electrochemical Sensor Array Combined with a Preconcentrator

A method for quantitative detection and discrimination of volatile sulfur compounds (VSCs) using an electrochemical sensor array combined with a preconcentrator was proposed. Halitosis is due to VSCs produced by bacterial metabolism inside the oral cavity. An organoleptic test is typically performed by a dental clinician for the assessment of halitosis, although it is a subjective test. Thus, an objective evaluation of halitosis is required. In this study, it was possible to discriminate among the VSCs such as hydrogen sulfide (H2S), methyl mercaptan (CH3SH), and dimethyl sulfide ((CH3)2S) over the range of 200ppb to 1000ppb. Moreover, mixture of two VSC vapors (H2S and CH3SH) at various mixing ratios were measured. The results indicated that the sensor responses to mixed samples satisfied the linear superposition. The mixture compositions of VSCs were almost correctly obtained from the sensor responses using partial least squares (PLS) regression analysis.

Fabrication of an Electrochemical Sensor Array for 2D H2O2 Imaging

To achieve accurate real-time 2D H2O2 distribution imaging, we fabricated an H2O2 sensor array by dispensing small quantities of enzyme solution very accurately onto electrodes in an electrode array. We confirmed the accuracy of the dispensing technique. We then investigated the responses of each sensor to H2O2 injection using a 64-channel multipotentiostat and found they varied by 20%. Real-time 2D H2O2 imaging was therefore successfully realized by using our flow cell system. This sensor array will be useful for the non-invasive, real-time monitoring of the H2O2 distribution in biological samples.

Smart Chemical Taste Sensor for Determination and Prediction of Taste Qualities Based on a Two-Phase Optimized Radial Basis Function Network

A novel, smart, chemical taste sensor that realistically mimics the behavior of the human gustatory system is described. The taste sensor consists of an array of electrochemical sensors that represent the gustatory receptors on the human tongue, and a two-phase optimized radial basis function network (RBFN) to represent the human brain, which comprehensively analyzes the gustatory stimulation and judges the overall taste. In the illustrated model, eight electrodes were fabricated to determine the eight major taste-causing substances, Na+, K+, Cl-, H+, sucrose, glucose, glutamate, and caffeine. The detected signals were fed to a two-phase RBFN optimized by the implementation of a basis optimization algorithm and weight decay term for appropriate data processing. The first phase of the two-phase RBFN quantifies the amount of taste-causing substances in food samples from the responses of the electrodes. These results are then fed to the second phase, which correlates the amount of substances with the overall taste. The final output is scored on a scale of 1-5 for each of the five basic tastes sensed by the human gustatory system, which are saltiness, sourness, sweetness, bitterness, and umami. The constructed network estimated the intensity of the basic tastes of 30 drink varieties with an average relative error of 7.0% compared to the human scores. The network could also estimate the variance in the human sensory perception. Moreover, the sensor successfully predicted the interactions of tastes such as suppression of bitterness by sweetness and enhancement of umami by saltiness, which are illusions sensed by the human gustatory system. With these abilities, the novel taste sensor can be considered as a quantitative yet humanlike sensor with a great potential for practical applications.

Hydrogen peroxide distribution and neuronal cell death in a rat hippocampal slice

Hydrogen peroxide (H 2O 2), a by-product of a degenerative reaction of reactive oxygen species in mammalian cells, has been examined in association with the pathology of neurodegenerative disorders. In this study, we monitor the H 2O 2 release from a rat hippocampal slice culture at multiple positions using our novel electrochemical sensor array. The H 2O 2 sensor array is fabricated by modifying a 64-channel ITO electrode array with an enzyme, horseradish peroxidase, and an electron transfer mediator. A hippocampal slice is placed on the array and the current at each sensor is monitored using a multipotentiostat. When we introduce kainate (KA) into the solution to stimulate the slice, the H 2O 2 concentration remained largely unchanged. In contrast, in the presence of 3-amino-1,2,4-triazol (3-AT), a catalase inhibitor, we can observe a distinct increase in the H 2O 2 concentration as a result of KA stimulation especially in the CA3 region in the hippocampus. KA exposure is also shown to cause neuronal cell death in the CA3 region by Cresyl Violet staining. These results reveal that our H 2O 2 sensor array can detect H 2O 2 released from neurons by KA stimulation under catalase inhibition and that this H 2O 2 is associated with neuronal cell death.

An electrochemical sensor array system for the direct, simultaneous in vitro monitoring of nitric oxide and superoxide production by cultured cells

A new approach for an amperometric array sensor platform employing arrays of sensors in a 24-well cell culture plate format has been developed for simultaneous in vitro determination of nitric oxide (NO) and superoxide free radicals (O(2)(-)) produced by stimulated cells. The work reported focuses on the direct, real-time monitoring of extracellular production of these two analytes, as well as the effects of their interaction. The sensor platform was manufactured by a combination of sputtering gold electrodes and screen-printing carbon electrodes. The O(2)(-) sensor uses covalent immobilization of cytochrome c via a binder, DTSSP (3,3'-dithio-bis(sulphosuccinimidylpropionate) onto the surface of the Au electrodes, whereas the NO sensor system involves an NiTSPc (nickel tetrasulfonated phthalocyanine) film electrodeposited onto the surface of the carbon electrodes and subsequently covered with an external layer of Nafion. For in vitro demonstration of the platforms as a potential drug-screening system, A172 glioblastoma cells were cultured and transferred into the 24-well arrays. Simultaneous and direct monitoring of NO and O(2)(-) production as a response to chemicals of biomedical relevance was carried out. The results obtained demonstrated that it would be possible to envisage a drug screening platform for compounds designed to be inhibitors of nitric oxide synthase or to have an inhibitory effect on superoxide free radical production. By suitable modification of the electrodes employed it would also be possible to extend the platform to measure alternative species.

Rapid, species-specific detection of uropathogen 16S rDNA and rRNA at ambient temperature by dot-blot hybridization and an electrochemical sensor array

Development of rapid molecular approaches for pathogen detection is key to improving treatment of infectious diseases. For this study, the kinetics and temperature-dependence of DNA probe hybridization to uropathogen species-specific sequences were examined. A set of oligonucleotide probes were designed based on variable regions of the 16S gene of the Escherichia coli, Proteus mirabilis, Klebsiella oxytoca, and Pseudomonas aeruginosa. A universal bacterial probe and probes-specific for gram-positive and gram-negative organisms were also included. The oligonucleotide probes discriminated among 16S genes derived from 11 different species of uropathogenic bacteria applied to nylon membranes in a dot-blot format. Significant binding of oligonucleotide probes to target DNA and removal of nonspecific binding by membrane washing could both be achieved rapidly, requiring as little as 10 min. An oligonucleotide probe from the same species-specific region of the E. coli 16S gene was used as a capture probe in a novel electrochemical 16-sensor array based on microfabrication technology. Sequence-specific hybridization of target uropathogen 16S rDNA was detected through horseradish peroxidase acting as an electrochemical transducer via a second, detector probe. The sensor array demonstrated rapid, species-specific hybridization in a time course consistent with the rapid kinetics of the dot-blot hybridization studies. As in the dot-blot hybridization studies, species-specific detection of bacterial nucleic acids using the sensor array approach was demonstrated both at 65 °C and at room temperature. These results demonstrate that molecular hybridization approaches can be adapted to rapid, room temperature conditions ideal for an electrochemical sensor array platform.

Electrochemical sensor array made from bisphthalocyanine modified carbon paste electrodes for discrimination of red wines

A new sensor system devoted to the discrimination of red wines is presented. It consists of an array of electrochemical sensors made from carbon paste electrodes modified with three rare-earth bisphthalocyaninate compounds, including lutetium(III), gadolinium(III) and praseodymium(III) bisphthalocyaninates. The sensor responses have been evaluated either by cyclic voltammetry or by square wave voltammetry, and demonstrate that such voltammetric sensors show a rich response when immersed in red wines. In addition, this response can be tuned by changing the central metal atom of the double-decker complexes, obtaining sensing units with high cross-selectivity. The sensor array is exposed to six Spanish red wines made from the same grape variety, prepared using a similar vinification method, but belonging to three different geographic origins and ageing stages. The voltammograms indicate that each sensing unit provides a particular response to each class of wine. Finally, the signals coming from each sensor are combined and the system’s discrimination capability is evaluated by using principal component analysis (PCA), where a windowed slicing pre-processing method has been developed to capture the information throughout the global response. The PCA score plots of the obtained data show the discrimination of the tested wines with a good-quality performance.

Fusion of Three Sensory Modalities for the Multimodal Characterization of Red Wines

This work represents the first attempt to develop a sensory system, specifically designed for the characterization of wines, which combines three sensory modalities: an array of gas sensors, an array of electrochemical liquid sensors, and an optical system to measure color by means of CIElab coordinates. This new analytical tool, that has been called "electronic panel," includes not only sensors, but also hardware (injection system and electronics) and the software necessary for fusing information from the three modules. Each of the three sensory modalities (volatiles, liquids, and color) has been designed, tested, and optimized separately. The discrimination capabilities of the system have been evaluated on a database consisting of six red Spanish wines prepared using the same variety of grape (tempranillo) but differing in their geographic origins and aging stages. Sensor signals from each module have been combined and analyzed using pattern recognition techniques. The results of this work show that the discrimination capabilities of the system are significantly improved when signals from each module are combined to form a multimodal feature vector.

Electrochemical approaches for chemical and biological analysis on Mars.

Obtaining in situ chemical data from planetary bodies such as Mars or Europa can present significant challenges. The one analytical technique that has many of the requisite characteristics to meet such a challenge is electroanalysis. Described here are three electroanalytical devices designed for in situ geochemical and biological analysis on Mars. The Mars Environmental Compatibility Assessment (MECA) was built and flight qualified for the now cancelled NASA Mars 2001 Lander. Part of MECA consisted of four "cells" containing arrays of electrochemical based sensors for measuring the ionic species in soil samples. A next-generation MECA, the Robotic Chemical Analysis Laboratory (RCAL), uses a carousel-type system to allow for greater customization of analytical procedures. A second instrument, proposed as part of the 2007 CryoScout mission, consists of a flow-through inorganic chemical analyzer (MICA). CryoScout is a torpedo-like device designed for subsurface investigation of the stratigraphic climate record embedded in Mars' north polar cap. As the CryoScout melts its way through the ice cap, MICA will collect and analyze the meltwater for a variety of inorganics and chemical parameters. By analyzing the chemistry locked in the layers of dust, salt, and ice, geologists will be able to determine the recent history of climate, water, and atmosphere on Mars and link it to the past. Finally, electroanalysis shows its abilities in the detection of possible microorganism on Mars or elsewhere in the solar system. To identify an unknown microorganism, one that may not even use Earth-type biochemistry, requires a detection scheme which makes minimal assumptions and looks for the most general features. Recent work has demonstrated that the use of an array of electrochemical sensors which monitors the changes in a solution via electrical conductivity, pH, and ion selective electrodes, can be used to detect minute chemical perturbations caused by the growth of bacteria and with the correct methodology provide unamibiguous detection of such life forms.

Electrochemical monitoring of glutamate release at multiple positions in a rat hippocampal slice.

The continuous monitoring of the distribution of glutamate (Glu), a neurotransmitter released at synaptic terminals, is important in terms of understanding the signal transfer mechanism in the brain. In this study, we monitored the concentration of Glu released at multiple positions in a hippocampal slice continuously, and obtained an approximate Glu distribution by using our electrochemical glutamate sensor array. After confirming our sensor's high sensitivity to Glu, we placed a slice on the array, and measured the currents at selected electrodes in the array. When we stimulated a specific position in the slice electrically, the glutamate concentration increased in different areas after several tens of seconds. The presence of glutamate receptor blockers suppressed these increases. This suggests that the electrical signal was transferred along with neurons through synapses and stimulated the Glu release. Our multichannel glutamate sensor should be a powerful tool to determining the distribution of real-time glutamate non-invasively for the studies using biological samples.

Case-based reasoning (CBR) for multicomponent analysis using sensor arrays: application to water quality evaluation.

This paper reports by the first time the use of a simple, convenient and improved method of artificial intelligence (case-based reasoning, CBR) for multicomponent analysis, involving the data treatment afforded by electrochemical sensor arrays. The suitability of this method has been shown in aqueous solutions containing mixtures of ions of different nature and concentration, being applied in this case to a rapid recognition and evaluation of mineral water samples chosen as an illustrative example. The convenience of such a procedure opens the way to dispose of a versatile tool applicable for obtaining analytical information about ions present in a wide range of other multicomponent systems, and also to ascribe properties inherent to the examined samples.