Impedimetric transducers based on interdigitated electrode arrays for bacterial detection - A review.

Application of the impedance spectroscopy technique to detection of bacteria has advanced considerably over the last decade. This is reflected by the large amount of publications focused on basic research and applications of impedance biosensors. Employment of modern technologies to significantly reduce dimension of impedimetric devices enable on-chip integration of interdigitated electrode arrays for low-cost and easy-to-use sensors. This review is focused on publications dealing with interdigitated electrodes as a transducer unit and different bacteria detection systems using these devices. The first part of the review deals with the impedance technique principles, paying special attention to the use of interdigitated electrodes, while the main part of this work is focused on applications ranging from bacterial growth monitoring to label-free specific bacteria detection.

Calcium-selective electrodes based on photo-cured polyurethane-acrylate membranes covalently attached to methacrylate functionalized poly(3,4-ethylenedioxythiophene) as solid-contact.

We report here the fabrication of solid-contact calcium-selective electrodes (Ca2+-SCISEs) made of a polyurethane acrylate ion-selective membrane (ISM) that was covalently attached to the underlying ion-to-electron transducer (solid-contact). Methacrylate-functionalized poly(3,4-ethylenedioxythiophene) (Meth-PEDOT) and Meth-PEDOT films containing either multiwalled carbon nanotubes (MWCNT) or carboxylated MWCNT (cMWCNT) were used as solid contacts. The solid contacts were deposited by drop-casting on screen-printed electrodes and characterized by cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS) and potentiometry. Covalent binding between the solid contact and the ISM was obtained via photopolymerization in order to increase the robustness of the Ca2+-SCISEs. The performance of the Ca2+-SCISEs was studied by measuring their potentiometric response and their sensitivity to light, oxygen and carbon dioxide. Meth-PEDOT was found to be a promising solid-contact material to develop low-cost and easy to prepare ISEs.

Solid contact ion sensor with conducting polymer layer copolymerized with the ion-selective membrane for determination of calcium in blood serum.

A new solid contact ion selective electrode with intermediate conducting polymer (CP) layer formed by electropolymerization on a gold electrode of a bifunctional monomer, n-phenyl-ethylenediamine-methacrylamide (NPEDMA), which contains a methacrylamide group attached to aniline, is presented. The conducting polymer was studied by means of optical spectroscopy, cyclic voltammetry and potentiometric measurements. Ca2+-ion-selective membrane based on acrylated urethane polymer was shown to co-polymerize with the CP forming highly adhesive boundary that prevents formation of water layers between the CP and membrane, thus enhancing the stability and life-time of the sensor. The designed ion-selective electrode was successfully used for determination of total calcium ion concentration in blood serum samples.

Impedimetric antimicrobial peptide-based sensor for the early detection of periodontopathogenic bacteria.

Peri-implantitis, an inflammation caused by biofilm formation, constitutes a major cause of implant failure in dentistry. Thus, the detection of bacteria at the early steps of biofilm growth represents a powerful strategy to prevent implant-related infections. In this regard, antimicrobial peptides (AMPs) can be used as effective biological recognition elements to selectively detect the presence of bacteria. Thus, the aim of the present study was to combine the use of miniaturized and integrated impedimetric transducers and AMPs to obtain biosensors with high sensitivity to monitor bacterial colonization. Streptococcus sanguinis, which is one of the most prevalent strains in the onset of periodontal diseases, was used as a model of oral bacteria. To this end, a potent AMP derived from human lactoferrin was synthesized and covalently immobilized on interdigitated electrode arrays (IDEA). X-ray photoelectron spectroscopy (XPS) and electrochemical impedance spectroscopy (EIS) were employed to optimize and characterize the method of immobilization. Noteworthy, the interaction of Streptococcus sanguinis with AMP-coated sensors provoked significant changes in the impedance spectra, which were univocally associated with the presence of bacteria, proving the feasibility of our method. In this regard, the developed biosensor permits to detect the presence of bacteria at concentrations starting from 10(1) colony forming units (CFU)mL(-1) in KCl and from 10(2) CFUmL(-1) in artificial saliva. Moreover, the system was devoid of cytotoxicity for human fibroblasts. These results indicate that the proposed approach can be effective in the detection of initial stages of biofilm formation, and may be useful in the early prevention and treatment of peri-implantitis.

Miniaturized metal oxide pH sensors for bacteria detection.

It is well known that the metabolic activity of some microorganisms results in changes of pH of the culture medium, a phenomenon that can be used for detection and quantification of bacteria. However, conventional glass electrodes that are commonly used for pH measurements are bulky, fragile and expensive, which hinders their application in miniaturized systems and encouraged to the search for alternatives. In this work, two types of metal oxide pH sensors have been tested to detect the metabolic activity of the bacterium Escherichia coli (E. coli). These pH sensors were produced on silicon chips with platinum metal contacts, onto which thin layers of IrOx or Ta2O5 were incorporated by two different methods (electrodeposition and e-beam sputtering, respectively). In order to facilitate measurement in small sample volumes, an Ag/AgCl pseudo-reference was also screen-printed in the chip and was assayed in parallel to an external Ag/AgCl reference electrode. As it is shown, the developed sensors generated results indistinguishable from those provided by a conventional glass pH-electrode but could be operated in significantly smaller sample volumes. After optimization of the detection conditions, the metal oxide sensors are successfully applied for detection of increasing concentrations of viable E. coli, with detection of less than 10(3)cfu mL(-1) in undiluted culture medium in just 5h.

Response of a microcapillary impedimetric transducer to changes in surface conductance at liquid/solid interface.

A new device based on an interdigitated electrode array with electrode digits located at the bottom of microcapillaries is presented. Microcapillaries formed in silicon dioxide are 3 µm wide, 4 µm high and are open at the top, so that in contact with an electrolyte solution the AC current flows from one capillary to another and is significantly affected by changes in surface conductivity at the SiO2/electrolyte interface. The effect of charged polyelectrolyte layers electrostatically assembled on the sensor surface on the surface conductivity in solutions with different KCl concentration is presented. From measured impedance spectra polyelectrolyte adsorption curve is determined. The device is shown to be useful for real time adsorption kinetics monitoring.

An impedimetric chemical sensor for determination of detergents residues.

A new impedimetric sensor based on an interdigitated electrode array with electrode digits located at the bottom of microcapillaries formed in silicon dioxide is presented. Microcapillaries are opened at the top, so that in contact with an electrolyte solution the ac current flows close to the surface of the capillary wall from one electrode to another and is significantly affected by changes in the surface conductance at the SiO2/electrolyte interface. Adsorption of detergents on the sensor surface affects the charge distribution in the electrical double layer and thus the surface conductance. These changes are registered by measuring impedance. Effect of surface adsorption of ionic and non-ionic surfactants on the sensor impedance is studied. The sensor is shown to be able to measure commercial detergents residues in a tap water starting from 5 ppm even in solutions with high electrolyte conductivity.

The influence of CO(2) on ISFETs with polymer membranes and characterization of a carbonate ion sensor.

The influence of CO(2) and acetic acid on the response of ISFET sensors with PVC and photocured polyurethane polymer membranes with valinomycin as an ionophore was assessed. Experimental results show that the presence of these compounds has no effect on sensors parameters even after prolonged soaking in a water solution. Using a photocured polyurethane polymer as an ion-selective membrane matrix for an ISFET, a carbonate ion sensor was developed with hexyl-p-trifluoroacetylbenzoate (HE) as an ionophore. Effect of cationic and anionic lipophilic additives on the sensors response was studied. Sensors with the optimized membrane composition based on HE (7.9%, w/w) and tridodecylmethylammonium chloride (5.7%, w/w) show sensitivity of 27-30mV per decade of carbonate ion concentration, sufficient selectivity in front of chloride ions, and a lifetime of 3-5 months.

Photocurable pH-sensitive membrane for ion-selective field effect transistors.

H(+)-ion sensitive ISFETs with photocured polyurethane-based polymer membranes with three different neutral carrier ionophores and four different plasticizers have been studied in 0.05 M TRIS-HCl solutions and in background solutions containing 140 mM of Na(+) ions. The optimised membrane composition showing the best selectivity contains 2.2-2.5 wt.% of the ionophore (tridodecylamine), 36-41 wt.% of a plasticizer, and only 10 mol% of a lipophilic salt KTClPhB. The optimal ionophore/lipophilic salt ratio obtained in this work differs significantly from theoretically recommended for pH-sensitive ion-selective membranes. It is assumed that this is due to the participation of the ionophore (tertiary amine) and lipophilic additives (tetrachlorophenylborate anion) in additional photochemical reactions occurring during irradiation of the membrane matrix. Sensors with the optimised membrane composition showed sufficient sensor selectivity in front of sodium ions for clinical and biomedical applications and the lifetime of more than 3 months.

Integrated multi-sensor chip with photocured polymer membranes containing copolymerized plasticizer for direct pH, potassium, sodium and chloride ions determination in blood serum.

An analytical system based on a sensor array with ion-selective field effect transistors (ISFETs) monolithically integrated in one chip covered with photocured polymer membranes containing copolymerized plasticizer and a sequential injection analysis (SIA) is shown to offer an automation of the analysis of blood serum components. For sequential injection system a custom made dual channel flow cell for the sensor array was developed. Optimisation of ion-sensitive membrane characteristics and calibration solution compositions were carried out. The system was used to analyze sodium, potassium, chloride ion contents in blood serum samples. The precision of the ion determination in samples was typical for potentiometric method with standard deviation of about 3-5%.

Three-dimensional interdigitated electrode array as a transducer for label-free biosensors.

A new transducer for biosensor applications has been developed based on a three-dimensional interdigitated electrode array (IDEA) with electrode digits separated by an insulating barrier. Binding of molecules to a chemically modified surface of the transducer induces important changes in conductivity between the electrodes. Three-dimensional sensor shows considerable improvement compared with a standard planar IDEA design. The potential of the developed device as a sensor transducer to detect immunochemical and enzymatic reactions, as well as DNA hybridization events is demonstrated. The immunosensor allows direct detection of the antibiotic sulfapyridine and shows the IC(50) parameter value of 5.6 microgL(-1) in a buffer. Immunochemical determination occurs under competitive configurations and without the use of any label. Each modified sensor is of a single use. Nevertheless, biochemical reagents can be easily cleaned off the sensor surface for its reuse. Layer-by-layer method of used to deposit polyethyleneimine and glucose oxidase showed that the sensor is also highly effective for detecting single and multilayered molecular assemblies.

Electronic integrated multisensor tongue applied to grape juice and wine analysis.

An integrated multisensor composed by six ISFET devices selective to common ions and heavy metals combined with a flow injection analysis (FIA) system has been applied as an electronic tongue to grape juice and wine sample analysis. The data obtained for several grape and wine variety samples analysis have been treated using multiparametric tools like principal component analysis (PCA) and soft independent modelling class analogy technique (SIMCA) for the patterning recognition and classification of samples and partial least squares (PLS) regression for quantification of several parameters of interest in wine production. The results obtained have demonstrated the potential of using those multisensors as electronic tongues not only for distinguishing the samples according to the grape variety and the vintage year but also for quantitative prediction of several sample parameters.

Lowering the detection limit of calcium selective ISFETs with polymeric membranes.

Detection limits of Ca sensitive ISFETs with photocured polyurethane membranes with a neutral carrier and an ion-exchanger as ionophores are studied. It is shown that conditioning of ISFETs in solutions containing only interfering ions results in reduction of the detection limit. Zero-current ion fluxes approach elaborated for polymeric membrane ion-selective electrodes (ISEs) is used to explain the obtained results.