Highly Sensitive Dopamine Detection Using a Bespoke Functionalised Carbon Nanotube Microelectrode Array

Understanding how the brain works requires developing advanced tools that allow measurement of bioelectrical and biochemical signals, including how they propagate between neurons. The introduction of nanomaterials as electrode materials has improved the impedance and sensitivity of microelectrode arrays (MEAs), allowing high quality recordings of single cells in situ using electrode diameters of ≤20 μm. MEAs also have the potential to measure electroactive biological molecules in situ, such as dopamine, a neurotransmitter in the nervous system. Thus, this work focused on fabricating a functionalised carbon nanotube (CNT)‐based MEA to demonstrate its potential for future measurement of small signals generated from excitable cells. To this end, the functionalised CNT MEA has recorded one of the lowest electrochemical interfacial impedances available in the literature, 2.8±0.2 kΩ, for an electrode of its geometric surface area. Electrochemical detection of dopamine revealed again one of the best sensitivity values per area available in the literature, 9.48 μA μM⁻¹ mm⁻². Additionally, a limit of detection of 7 nM was recorded for dopamine using the functionalised CNT MEA, with selectivity against common electrochemical interferents such as ascorbic acid. These results indicate improvement beyond currently available MEAs, along with the feasibility of using these devices for multi‐site detection of physiologically relevant electroactive biomolecules.     

Biological application of microelectrode arrays in drug discovery and basic research

Electrical activity of electrogenic cells in neuronal and cardiac tissue can be recorded by means of microelectrode arrays (MEAs) that offer the unique possibility for non-invasive extracellular recording from as many as 60 sites simultaneously. Since its introduction 30 years ago, the technology and the related culture methods for electrophysiological cell and tissue assays have been continually improved and have found their way into many academic and industrial laboratories. Currently, this technology is attracting increased interest owing to the industrial need to screen selected compounds against ion channel targets in their native environment at organic, cellular, and sub-cellular level.As the MEA technology can be applied to any electrogenic tissue (i.e., central and peripheral neurons, heart cells, and muscle cells), the MEA biosensor is an ideal in vitro system to monitor both acute and chronic effects of drugs and toxins and to perform functional studies under physiological or induced pathophysiological conditions that mimic in vivo damages. By recording the electrical response of various locations on a tissue, a spatial map of drug effects at different sites can be generated, providing important clues about a drug's specificity.In this survey, examples of MEA biosensor applications are described that have been developed for drug screening and discovery and safety pharmacology in the field of cardiac and neural research. Additionally, biophysical basics of recording and concepts for analysis of extracellular electrical signals are presented.Abbreviations AP action potential - DG dentate gyrus - EC entorhinal cortex - ECG electrocardiogram - ERG electroretinogram - LFP local field potentials - MEA microelectrode array - PSTH peri-stimulus–time histogram - SNR signal-to-noise ratio     

Improvement of the proton exchange membrane fuel cell performances by optimization of the hot pressing process for membrane electrode assembly

The present study deals with PEM fuel cells, namely with the optimization of the hot pressing process for membrane electrode assembly (MEA) fabrication. Designs of experiments (DoE) have been used for evaluating the effect of hot pressing parameters (pressure, temperature, and time) on the MEA electrical performances. Full factorial 2³ DoE showed that the most important parameter is the pressing temperature. Surface response methodology indicated a non-monotonous behavior of the MEA electrical performances with respect to the pressing temperature. The MEA electrical performances increased with the pressing temperature in the temperature range from 100 to 115 °C, and decreased significantly in the temperature range from 115 to 130 °C. This behavior was attributed to drastic changes of the Nafion® 112 membrane properties and membrane/electrode interface over this temperature range. Observations of the MEA cross-section structure by scanning electron microscopy confirmed such hypotheses. Thermo-mechanical properties of Nafion® as determined by dynamic scanning calorimetry allowed estimating the glass transition temperature at ca. T _g?≈?117 °C in the conditions of the present study. The higher H₂/air fuel cell performance of ca. 0.8 W cm^?2 was obtained with the optimized pressing temperature for MEA fabrication of ca. 115 °C close to the T _g temperature of Nafion® 112, whereas for higher temperature the structure of the Nafion® membrane and of the membrane–electrode interface is damaged.     

Improved conversion efficiency in dye-sensitized solar cells based on electrospun Al-doped ZnO nanofiber electrodes prepared by seed layer treatment

The application of electrospun nanofibers in electronic devices is limited due to their poor adhesion to conductive substrates. To improve this, a seed layer (SD) is introduced on the FTO substrate before the deposition of the electrospun composite nanofibers. This facilitates the release of interfacial tensile stress during calcination and enhances the interfacial adhesion of the AZO nanofiber films with the FTO substrate. Dye-sensitized solar cells (DSSC) based on these AZO nanofiber photoelectrodes have been fabricated and investigated. An energy conversion efficiency (η) of 0.54-0.55% has been obtained under irradiation of AM 1.5 simulated sunlight (100 mW/cm²), indicating a massive improvement of η in the AZO nanofiber film DSSCs after SD-treatment of the FTO substrate as compared to those with no treatment. The SD-treatment has been demonstrated to be a simple and facile method to solve the problem of poor adhesion between electrospun nanofibers and the conductive substrate.     

Towards a multi-electrode array (MEA) system for patterned neural networks

Here we describe the development of a unique multi-electrode array (MEA) system capable of supporting patterned neural networks in vitro. The significant features of this BioMEMS system are: (1) precise and exclusive positioning of the neural cell bodies to the electrode region, (2) definition of interconnecting pathways among the neurons by using patterned self-assembled monolayers (SAMs), and (3) one-to-one stimulation and recording from precisely positioned neurons. We present for the first time the design and fabrication of an MEA chip supporting patterned neuronal networks, as well as the initial electrical recordings from two types of neurons.     

Improved interfacial adhesion of membrane electrode assemblies using polymer binders and pore-filling membranes in alkaline membrane fuel cells

The development of alkaline membrane fuel cells (AMFCs) will enable the use of non-platinum catalysts and hydrocarbon-based electrolyte membranes. Such catalysts are intrinsically stable and have activities similar to that of platinum in an alkaline environment. A pore-filling membrane has been made from a porous, high-density polyethylene substrate to fabricate durable, AMFC membrane electrode assemblies (MEAs). Because of the low binding ability of the hydrocarbon ionomer in the preparation of AMFC MEAs, polymer binders were added to the catalyst slurries to form a firmly bound interface. A content of 20 wt% polyethylene (PE) binder, the same material as the porous substrate in the pore-filling membrane, exhibits the best attachment of the non-platinum catalyst particles to the pore-filling, hydrocarbon anion-exchange membranes. The addition of a PE binder improves adhesion at the MEA interface and diminished contact resistance. The improved durability of the MEA is confirmed by continuous, constant-voltage operation. Adhesion between the cathode catalyst layer and the pore-filling membrane is also investigated after mild hot-pressing to test the use of decal method in the fabrication of AMFCs. The catalyst layer with the PE binder was completely transferred to the pore-filling membrane at 50 °C and 30 bar?cm^?2, but not for the PTFE binder.     

A study of the mechanism of response of liquid ion exchanger calcium selective electrodes: Part III. Membrane behaviour under non-zero current conditions

The I-E response of the liquid membrane of the calcium selective electrode is studied under constant or linearly varying current and voltage. An increase in the membrane resistance, recorded when an electrical current crosses the membrane, is due to the outflow of Cl^? ions initially present in the membrane. When calcium ions are replaced by alkaline ions inside the membrane at constant current, the decrease of the membrane resistance due to an ion exchange is in agreement with the conductivity measurements (Part II). When the applied voltage is imposed besides the ion exchange one must take into account the interfacial overpotential to explain the important rectification effect observed. The interfacial transfer constant rate of alkaline ions seems greater than that of Ca²⁺ ion.     

Ultrasensitive bioelectronic devices based on conducting polymers for electrophysiology studies

Conducting polymer electrodes based on poly(3,4-ethylenedioxythiophene doped with poly(styrenesulfonate) (PEDOT:PSS) are evaluated as transducers to record extracellular signals in cell populations. The performance of the polymer electrode is compared with a gold electrode. A small-signal impedance analysis shows that in the presence of an electrolyte, the polymer electrode establishes for frequencies below 100 Hz a higher capacitive electrical double layer at the electrode/electrolyte interface. Furthermore, the polymer/electrolyte interfacial resistance is several orders of magnitude lower than the resistance of the gold/electrolyte interface. The polymer low interfacial resistance minimizes the intrinsic thermal noise and increases the system sensitivity. The ultra-sensitivity of the polymer-based transducer system was demonstrated by recording the electrical activity of cancer cells of the nervous system.     

CMOS-Based Bio/Chemosensor and Bioelectronic Microsystems

Microfabrication techniques and, in particular, complementary-metal-oxide-semiconductor (CMOS) technology have been used to devise chemo/biosensors [1-3] as well as bioelectronic microsystems [4-7]. Examples of micromachined bio- or chemosensors, such as cantilevers or micoelectrode arrays, will be shown, and the electrical interfacing of CMOS microelectronics with biological entities or electrogenic cells, i.e., cells that react upon electrical stimulation and, in turn, produce electrical signals (heart cells or neurons) are detailed. CMOS-based, fully integrated microelectrode arrays for bidirectional communication (stimulation and recording) with electrogenic cells are presented. These devices are capable of monitoring relevant electrophysiological responses of cells to electrical stimuli or to pharmacological agents with prospective applications in the field of bio-inspired information processing or pharmascreening.     

Electrically controlling cell adhesion,growth and migration

We have developed a neurochip to control the adhesion and outgrowth of individual neurons by electrochemical removal of protein repellent molecules from transparent electrodes. The neurochip architecture is based on three parallel indium-tin-oxide (ITO) electrodes on a SiO₂ substrate and a photoresist structure forming a landing spot for the neuron soma and two lateral outgrowth pathways for the neurites. The whole surface was turned protein and cell repellent with poly(ethylene glycol) grafted-poly(l-lysine) (PLL-g-PEG) before enabling neuron soma adhesion by selective PLL-g-PEG removal. After the neuron has settled down a potential was applied to the pathway electrodes to permit the neurite outgrowth along pathways formed by the SU8 structure. We also show the possibility to control cell migration by small pulsed currents. Myoblasts were therefore seeded on a chemical pattern of cell adhesive PLL and cell resistant PLL-g-PEG. The PLL-g-PEG was then removed electrochemically from the electrodes to permit migration onto the cell free electrodes. Electrodes without applied current were confluently overgrown within 24 h but a small pulsed current was able to inhibit cell growth on the bare ITO electrode for more than 72 h. With both techniques, cell adhesion, growth and migration can be controlled dynamically after the cells started to grow on the substrate. This opens new possibilities: we believe the key to control the development of topologically controlled neuron networks or more complex co-cultures is the combination of passive surface modifications and active control over the surface properties at any time of the experiment.     

Electrode interfaces switchable by physical and chemical signals for biosensing, biofuel, and biocomputing applications

This review outlines advances in designing modified electrodes with switchable properties controlled by various physical and chemical signals. Irradiation of the modified electrode surfaces with various light signals, changing the temperature of the electrolyte solution, application of a magnetic field or electrical potentials, changing the pH of the solutions, and addition of chemical/biochemical substrates were used to change reversibly the electrode activity. The increasing complexity in the signal processing was achieved by integration of the switchable electrode interfaces with biomolecular information processing systems mimicking Boolean logic operations, thus allowing activation and inhibition of electrochemical processes on demand by complex combinations of biochemical signals. The systems reviewed range from simple chemical compositions to complex mixtures modeling biological fluids, where the signal substrates were added at normal physiological and elevated pathological concentrations. The switchable electrode interfaces are considered for future biomedical applications where the electrode properties will be modulated by the biomarker concentrations reflecting physiological conditions.

High performance nitrile copolymers for polymer electrolyte membrane fuel cells

This paper reports the fuel cells (DMFC and PEMFC) performance using sulfonated poly(arylene ether ether nitrile) (SPAEEN) copolymers containing sulfonic acid group arranged in structurally different ways. The membrane electrode assembly (MEA) fabricated from SPAEEN containing 60 mol% of angled naphthalenesulfonic acid group (m-SPAEEN-60) had superior performance over those derived from pendent naphthalenesulfonic acid group (p-SPAEEN) or sulfonated hydroquinone (HQ-SPAEEN) in H₂/air and/or DMFC conditions. For example, the current density of the MEA using m-SPAEEN-60 at 0.5 V and 2.0 M methanol was 250 mA/cm², whereas the current densities of the MEAs using p-SPAEEN-50 and HQ-SPAEEN-56 were 185 and 190 mA/cm², respectively. In addition, compared with the sulfonated polysulfone (BPSH-35) and Nafion membranes, the copolymer containing nitrile group showed the improved cell performance. For example, the power density of the MEA using m-SPAEEN-60 at 250 mA/cm² and 2.0 M methanol was 125 mW/cm², whereas the power densities of the MEAs using sulfonated polysulfone (BPSH-35) and Nafion were 115 and 113 mW/cm², respectively. m-SPAEEN-60 showed stable cell performance during extended operation (>100 h).     

Photoactivation of a substrate for cell adhesion under standard fluorescence microscopes

Cell-culturing substrates where cell adhesion can be switched on by external stimuli during cell cultivation are useful scaffolds for tissue engineering, cell-based drug screening, and fundamental cellular studies. Here, we show a new strategy for photoactivation of a substrate for cell adhesion under standard fluorescence microscopes. A glass substrate chemically modified with an alkylsiloxane having a photocleavable 2-nitrobenzyl group was coated with bovine serum albumin to prevent cell adhesion. Upon irradiation under a fluorescence microscope, the protein was replaced with fibronectin, which made the irradiated region cell-adhesive. Subsequent seeding of HEK293 or COS7 cells produced patterns corresponding to the irradiated patterns. We succeeded for the first time in positioning single cells in proximity to cultivating single cells. The present method provides a general strategy for positioning single cells of same or different types at any locations on the substrate and will be useful for studying cell-cell interactions.     

Electrochemical impedance spectroscopy study of proteolysis using unmodified gold nanoparticles

Proteases are involved in numerous cell functions and abnormal proteolysis may lead to a diversity of serious diseases. Herein, a simple electrochemical method is developed to study proteolysis by employing unmodified gold nanoparticles (AuNPs). Substrate of a protease is modified on a gold disk electrode, forming a barrier for electrochemical species and reflecting a significant charge transfer resistance (R_ct). After the proteolysis process, the substrate can be cleaved coupled with the decline of R_ct. The electrical properties of the substrate residues on the electrode may also change, leading to the subsequent adsorption of AuNPs. Due to the excellent electrical conductivity of AuNPs, R_ct can be further decreased, which can be used to reveal the proteolysis process. The proposed method allows the determination of the model protease, trypsin, with desirable sensitivity and specificity. It may also hold great potential use in the study of other proteolysis processes and some biomedical applications in the future.     

Application of the avidin–biotin interaction to immobilize DNA in the development of electrochemical impedance genosensors

Impedance spectroscopy is a rapidly developing technique for the transduction of biosensing events at the surface of an electrode. The immobilization of biomaterial as DNA strands on the electrode surface alters the capacitance and the interfacial electron transfer resistance of the conductive electrodes. The impedimetric technique is an effective method of probing modifications to these interfacial properties, thus allowing the differentiation of hybridization events. In this work, an avidin bulk-modified graphite–epoxy biocomposite (Av-GEB) was employed to immobilize biotinylated oligonucleotides as well as double-stranded DNA onto the electrode surface. Impedance spectra were recorded to detect the change in the interfacial electron transfer resistance (R _et) of the redox marker ferrocyanide/ferricyanide at a polarization potential of +0.17 V. The sensitivity of the technique and the good reproducibility of the results obtained with it confirm the validity of this method based on a universal affinity biocomposite platform coupled with the impedimetric technique.     

Charge displacement by adhesion and spreading of a cell

The potentiostatic control of surface charge density and interfacial tension of an electrode immersed in an aqueous electrolyte solution offers a possibility for direct studies of non-specific interactions in cell adhesion. Unicellular marine alga, Dunaliella tertiolecta (Chlorophyceae) of micrometer size and flexible cell envelope was used as a model cell and 0.1 M NaCl as supporting electrolyte. The dropping mercury electrode acted as in situ adhesion sensor and the electrochemical technique of chronoamperometry allowed measurement of the spread cell-electrode interface area and the distance of the closest approach of a cell. The adhesion and spreading of a single cell at the mercury electrode causes a displacement of counter-ions from the electrical double layer over a broad range of the positive and negative surface charge densities (from +16.0 to -8.2 microC/cm2). The flow of compensating current reflects the dynamics of adhesive contact formation and subsequent spreading of a cell. The adhesion and spreading rates are enhanced by the hydrodynamic regime of electrode's growing fluid interface. The distance of the closest approach of an adherent cell is smaller or equal to the distance of the outer Helmholz plane within the electrical double layer, i.e. 0.3-0.5 nm. There is a clear evidence of cell rupture for the potentials of maximum attraction as the area of the contact interface exceeded up to 100 times the cross-section area of a free cell.     

Enhanced response of microbial fuel cell using sulfonated poly ether ether ketone membrane as a biochemical oxygen demand sensor

The present study is focused on the development of single chamber microbial fuel cell (SCMFC) using sulfonated poly ether ether ketone (SPEEK) membrane to determine the biochemical oxygen demand (BOD) matter present in artificial wastewater (AW). The biosensor produces a good linear relationship with the BOD concentration up to 650 ppm when using artificial wastewater. This sensing range was 62.5% higher than that of Nafion^®. The most serious problem in using MFC as a BOD sensor is the oxygen diffusion into the anode compartment, which consumes electrons in the anode compartment, thereby reducing the coulomb yield and reducing the electrical signal from the MFC. SPEEK exhibited one order lesser oxygen permeability than Nafion^®, resulting in low internal resistance and substrate loss, thus improving the sensing range of BOD. The system was further improved by making a double membrane electrode assembly (MEA) with an increased electrode surface area which provide high surface area for electrically active bacteria.     

Anode Catalysts for Direct Hydrazine Fuel Cells: From Laboratory Test to an Electric Vehicle

Novel highly active electrocatalysts for hydrazine hydrate fuel cell application were developed, synthesized and integrated into an operation vehicle prototype. The materials show in both rotating disc electrode (RDE) and membrane electrode assembly (MEA) tests the world highest activity with peak current density of 16 000 A g^?1 (RDE) and 450 mW cm^?2 operated in air (MEA).     

Development of high-performance cathodes for IT-SOFCs through beneficial interfacial reactions

We propose a new way to develop high-performance cathodes for IT-SOFCs by utilizing the interfacial reactions. SrCoO_x was selected as the starting electrode material, which took a vacancy-ordered 2H BaNiO₃-type structure and showed negligible ionic conductivity and low electrical conductivity. Phase reactions between SrCoO_x and Sm_0.2Ce_0.8O_1.9 happened at 900 °C or higher, resulting in the incorporation of Sm and Ce into its lattice structure. This promoted the phase transition to a cubic perovskite and led to substantial increase in the electrical conductivity and oxygen mobility of the electrode. By utilizing such phase reactions, the SrCoO_x + Sm_0.2Ce_0.8O_1.9 composite was developed into a high performance electrode with a low area specific resistance of 0.08 Ω cm^?2 at 650 °C. An anode-supported cell with such electrode delivered a peak power density of 795 mW cm^?2 at 600 °C.     

EC-STM observation on electrochemical response of fluidic phospholipid monolayer on Au(1 1 1) modified with 1-octanethiol

Electrochemical scanning tunneling microscopy (EC-STM) was applied to observe phospholipid layers over thiol-modified gold substrates as a model biological cell membrane. On a monolayer of 1-octanethiol on Au (1 1 1), a synthetic lipid, 1,2-dihexanoyl-sn-glycero-3-phosphocholine, was introduced in a neutral 0.05 M NH₄ClO₄ buffer solution. The lipid molecules formed a fluidic layer at 0.0 V vs. RHE of the substrate electrode potential. By cycling the electrode potential between +0.2 V and −0.2 V, the lipid layer reversibly changed over between the fluidic phase and a striped/grainy structure. This structural change might involve partial decomposition and oligomerization of phospholipids. This method will contribute for molecular biology by revealing the nanometer-scale structure of cell membrane.