Xavier Muñoz-Berbel

Label-free impedimetric immunosensor for sensitive detection of ochratoxin A

A novel label-free electrochemical impedimetric immunosensor for sensitive detection of ochratoxin A (OTA) was reported. A two-step reaction protocol was elaborated to modify the gold electrode. The electrode was first derivatized by electrochemical reduction of in situ generated 4-carboxyphenyl diazonium salt (4-CPDS) in acidic aqueous solution yielded stable 4-carboxyphenyl (4-CP) monolayer. The ochratoxin A antibody was then immobilized making use of the carbodiimide chemistry. The steps of the immunosensor elaboration and the immunochemical reaction between ochratoxin A and the surface-bound antibody were interrogated using cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The impedance change, due to the specific immuno-interaction at the immunosensor surface was utilized to detect ochratoxin A. The increase in electron-transfer resistance (DeltaR(et)) values was linearly proportional to the concentration of OTA in the range of 1-20ngmL(-1), with a detection limit of 0.5ngmL(-1).

Impedimetric approach for quantifying low bacteria concentrations based on the changes produced in the electrode-solution interface during the pre-attachment stage.

This paper describes an approach for quantifying low concentrations of bacteria, particularly Escherichia coli, based on the measurement of the initial attachment of bacteria to platinum surfaces, using impedance spectroscopy. The value of the interface capacitance in the pre-attachment stage (before 1min of attachment) showed correlation with suspended concentration of bacteria from 10(1) to 10(7)CFUmL(-1) (colony forming units per mL). This method was found to be sensitive to the attachment time, to the applied potential and to the size of the counter electrode. The sensor lifetime was also evaluated.

Characterization of fibrous polymer silver/cobalt nanocomposite with enhanced bactericide activity

This manuscript describes the synthesis (based on the intermatrix synthesis (IMS) method), optimization, and application to bacterial disinfection of Ag@Co polymer-metal nanocomposite materials with magnetic and bactericidal properties. This material showed ideal bactericide features for being applied to bacterial disinfection of water, particularly (1) an enhanced bactericidal activity (when compared with other nanocomposites only containing Ag or Co nanoparticles), with a cell viability close to 0% for bacterial suspensions with an initial concentration below 10(5) colony forming units per milliliter (CFU/mL) after a single pass through the material, (2) capacity of killing a wide range of bacterial types (from coliforms to gram-positive bacteria), and (3) a long performance-time, with an efficiency of 100% (0% viability) up to 1 h of operation and higher than 90% during the first 24 h of continuous operation. The nanocomposite also showed a good performance when applied to water samples from natural sources with more complex matrices with efficiencies always higher than 80%.

On-chip impedimetric detection of bacteriophages in dairy samples.

Bacteriophage infection of starter cultures constitutes a major problem in the dairy fermentation industry, which may bring about important economic losses. In this study, a rapid detection method of bacteriophages was developed based on analysis of impedance changes occurred upon infection of a host-biofilm established onto metal microelectrodes. Bacteriophage PhiX174 and Escherichia coli WG5 were chosen as models for bacteriophage and host strain, respectively, because of their easiness of manipulation. Impedimetric changes occurring at the microelectrode surface, caused by bacteriophage infection and subsequent lysis of the host strain, were monitored over a 6-h period after the initial inoculation of phages by non-faradic impedance spectroscopy (IS) in PBS and milk samples. Analysis of data was performed by two different approaches: (1) the equivalent circuit modelling theory, where a decrease in the magnitude of both the double layer and the biofilm capacitances due to the bacteriophage infection process was recorded, and (2) analysis of the impedance value, specially the impedance imaginary component (Z(i)) at selected frequencies. Z(i) is related to the capacitance of the circuit and also showed a decrease with respect to the control sample (without bacteriophages). The simplicity of the assay and the possibility of miniaturization of the system as well as its wide application, being able of detecting any bacteriophage as long as a suitable bacterial host is available, increase the number of applications to which this system could be used for.

Photonic Lab-on-a-Chip: Integration of Optical Spectroscopy in Microfluidic Systems

The integration of micro-optical elements with microfluidics leads to the highly promising photonic lab-on-a-chip analytical systems (PhLoCs). In this work, we re-examine the main principles which are underneath the on-chip spectrophotometric detection, approaching the PhLoC concept to a nonexpert audience.

Dual photonic-electrochemical lab on a chip for online simultaneous absorbance and amperometric measurements.

A dual lab on a chip (DLOC) approach that enables simultaneous optical and electrochemical detection working in a continuous flow regime is presented. Both detection modes are integrated for the first time into a single detection volume and operate simultaneously with no evidence of cross-talk. The electrochemical cell was characterized amperometrically by measuring the current in ferrocyanide solutions at +0.4 V vs gold pseudoreference electrode, at a flow rate of 200 μL min(-1). The experimental results for ferrocyanide concentrations ranging from 0.005 to 2 mM were in good agreement with the values predicted by the Levich equation for a microelectrode inside a rectangular channel, with a sensitivity of 2.059 ± 0.004 μA mM(-1) and a limit of detection (LoD) of (2.303 ± 0.004) × 10(-3) mM. Besides, optical detection was evaluated by measuring the absorbance of ferricyanide solutions at 420 nm. The results obtained therein coincide with those predicted by the Beer-Lambert law for a range of ferricyanide concentrations from 0.005 to 0.3 mM and showed an estimated LoD of (0.553 ± 0.001) × 10(-3) mM. The DLOC was finally applied to the analysis of L-lactate via a bienzymatic reaction involving lactate oxidase (LOX) and horseradish peroxidase (HRP). Here, the consumption of the reagent of the reaction (ferrocyanide) was continuously monitored by amperometry whereas the product of the reaction (ferricyanide) was recorded by absorbance. The DLOC presented good performance in terms of sensitivity and limit of detection, comparable to other fluidic systems found in the literature. Additionally, the ability to simultaneously quantify enzymatic reagent consumption and product generation confers the DLOC a self-verifying capability which in turn enhances its robustness and reliability.

Impedance-Based Biosensors for Pathogen Detection

Electrochemical impedance spectroscopy (EIS) is an important detection technique for biosensors. In the field of immunosensors, and particularly pathogen detection, it is one of the preferred electrochemical techniques because it does away with the use of enzyme labels or redox mediators. This chapter provides an introduction to the fundamentals of EIS and basic data analysis, with an emphasis on the most common features found in immunosensors and possible experimental limitations.

Monolithically integrated biophotonic lab-on-a-chip for cell culture and simultaneous pH monitoring

A poly(dimethylsiloxane) biophotonic lab-on-a-chip (bioPhLoC) containing two chambers, an incubation chamber and a monitoring chamber for cell retention/proliferation and pH monitoring, respectively, is presented. The bioPhLoC monolithically integrates a filter with 3 μm high size-exclusion microchannels, capable of efficiently trapping cells in the incubation chamber, as well as optical elements for real-time interrogation of both chambers. The integrated optical elements made possible both absorption and dispersion measurements, which were comparable to those made in a commercially available cuvette. The size-exclusion filter also showed good and stable trapping capacity when using yeast cells of variable size (between 5 and 8 μm diameter). For cell culture applications, vascular smooth muscle cells (VSMC), with sizes between 8 and 10 μm diameter, were used as a mammalian cell model. These cells were efficiently trapped in the incubation chamber, where they proliferated with a classical spindle-shaped morphology and a traditional hill-and-valley phenotype. During cell proliferation, pH changes in the culture medium due to cell metabolism were monitored in real time and with high precision in the monitoring chamber without interference of the measurement by cells and other (cell) debris.

Intermatrix synthesis of monometallic and magnetic metal/metal oxide nanoparticles with bactericidal activity on anionic exchange polymers

In this communication, the synthesis of nanoparticles on anionic exchange polymers by the Intermatrix Synthesis method is reported. Monometallic (Ag) and core–shell metal/metal oxide (Ag@Fe3O4) nanocomposites were synthesized and characterized. Their magnetic and bactericidal activities were evaluated.

Resolution of binary mixtures of microorganisms using electrochemical impedance spectroscopy and artificial neural networks

This work describes the resolution of binary mixtures of microorganisms using electrochemical impedance spectroscopy (EIS) and artificial neural networks (ANNs) for the processing of data. Pseudomonas aeruginosa, Staphylococcus aureus and Saccharomyces cerevisiae were chosen as models for Gram-negative bacteria, Gram-positive bacteria and yeasts, respectively. In this study, best results were obtained when entering the imaginary component of the impedance at each frequency (strongly related to the capacitive elements of the electrical equivalent circuit) into backpropagation neural networks made up by two hidden layers. The optimal configuration of these layers respectively used the radbas and the logsig transfer functions with 4 or 6 neurons in the first hidden layer and 10 neurons in the second one. In all cases, good prediction ability was obtained with correlation coefficients better than 0.989 when comparing the predicted and the expected values for a set of six external test samples not used in the training process.