Design and development of novel screen-printed microelectrode and microbiosensor arrays fabricated using ultrafast pulsed laser ablation

© 2016 Elsevier B.V. All rights reserved. A new generic platform for the development of microbiosensors combining screen-printing and ultrafast pulsed laser technologies has been developed, characterised and evaluated. This new platform consists of a layer of screen-printed carbon ink containing the enzyme and mediator, covered with an insulating layer formed from a dielectric screen printed ink. Microholes were drilled through the insulated layer by ultrafast pulsed laser ablation to generate the microbiosensor array. The geometry of the microelectrode array was evaluated by optical microscopy, white light surface profiling and scanning electron microscopy. The electrochemical behaviour of the microelectrode array was characterised by cyclic voltammetry and compared with macroelectrodes. The analytical performance of the microbiosensor array was evaluated with external counter and reference electrodes for hydrogen peroxide and glucose determination showing linearity up to 4 mmol L-1 and 20 mmol L-1 (360 mg dL-1) respectively. The full screen printed three-electrode configuration shows linearity for glucose determination up to 20 mmol L-1 (360 mg dL-1). This study provides a new fabrication method for microelectrode and microbiosensor arrays capable for the first time to retain the activity of the enzymatic system after processing by pulse laser ablation

At clinically relevant concentrations the anaesthetic/amnesic thiopental but not the anticonvulsant phenobarbital interferes with hippocampal sharp wave-ripple complexes

<p>Abstract</p> <p>Background</p> <p>Many sedative agents, including anesthetics, produce explicit memory impairment by largely unknown mechanisms. Sharp-wave ripple (SPW-R) complexes are network activity thought to represent the neuronal substrate for information transfer from the hippocampal to neocortical circuits, contributing to the explicit memory consolidation. In this study we examined and compared the actions of two barbiturates with distinct amnesic actions, the general anesthetic thiopental and the anticonvulsant phenobarbital, on in vitro SPW-R activity.</p> <p>Results</p> <p>Using an in vitro model of SPW-R activity we found that thiopental (50–200 μM) significantly and concentration-dependently reduced the incidence of SPW-R events (it increased the inter-event period by 70–430 %). At the concentration of 25 μM, which clinically produces mild sedation and explicit memory impairment, thiopental significantly reduced the quantity of ripple oscillation (it reduced the number of ripples and the duration of ripple episodes by 20 ± 5%, n = 12, <it>P </it>< 0.01), and suppressed the rhythmicity of SPWs by 43 ± 15% (n = 6, <it>P </it>< 0.05). The drug disrupted the synchrony of SPWs within the CA1 region at 50 μM (by 19 ± 12%; n = 5, <it>P </it>< 0.05). Similar effects of thiopental were observed at higher concentrations. Thiopental did not affect the frequency of ripple oscillation at any of the concentrations tested (10–200 μM). Furthermore, the drug significantly prolonged single SPWs at concentrations ≥50 μM (it increased the half-width and the duration of SPWs by 35–90 %). Thiopental did not affect evoked excitatory synaptic potentials and its results on SPW-R complexes were also observed under blockade of NMDA receptors. Phenobarbital significantly accelerated SPWs at 50 and 100 μM whereas it reduced their rate at 200 and 400 μM. Furthermore, it significantly prolonged SPWs, reduced their synchrony and reduced the quantity of ripples only at the clinically very high concentration of 400 μM, reported to affect memory.</p> <p>Conclusion</p> <p>We hypothesize that thiopental, by interfering with SPW-R activity, through enhancement of the GABA_Areceptor-mediated transmission, affects memory processes which involve hippocampal circuit activation. The quantity but not the frequency of ripple oscillation was affected by the drug.</p

Amperometric lactate biosensor for flow injection analysis based on a screen-printed carbon electrode containing Meldola's Blue-Reinecke salt, coated with lactate dehydrogenase and NAD+

A biosensor for the measurement of lactate in serum has been developed, which is based on a screen-printed carbon electrode, modified with Meldola's Blue-Reinecke Salt (MBRS-SPCE), coated with the enzyme lactate dehydrogenase NAD+ dependent (from Porcine heart), and NAD+. A cellulose acetate layer was deposited on the top of the device to act as a permselective membrane. The biosensor was incorporated into a commercially available, thin-layer, amperometric flow cell operated at a potential of only +0.05 V vs. Ag/AgCl. The mobile phase consisted of 0.2 M phosphate buffer pH 10 containing 0.1 M potassium chloride solution; a flow rate of 0.8 ml min-1 was used throughout the investigation. The biosensor response was linear over the range 0.55-10 mM lactate; the former represents the detection limit. The precision of the system was determined by carrying out 10 repeat injections of 10 mM l(+)lactic acid standard; the calculated coefficient of variation was 4.28%. It was demonstrated that this biosensor system could be applied to the direct measurement of lactate in serum without pre-treatment; therefore, this would allow high throughput-analysis, at low cost, for this clinically important analyte. © 2010 Elsevier B.V. All rights reserved

A flow injection system, comprising a biosensor based on a screen-printed carbon electrode containing Meldola's Blue-Reinecke salt coated with glucose dehydrogenase, for the measurement of glucose

A biosensor for the measurement of glucose in serum has been developed, based on a screen-printed carbon electrode modified with Meldola's Blue-Reinecke salt, coated with the enzyme glucose dehydrogenase (from Bacillus sp.), and nicotinamide adenine dinucleotide coenzyme (NAD+). A cellulose acetate layer was deposited on top of the device to act as a permselective membrane. The biosensor was incorporated into a commercially available, thin-layer, amperometric flow cell operated at a potential of only +0.05 V versus Ag/AgCl. The mobile phase consisted of 0.2 M phosphate buffer (pH 7.0) containing 0.1 M potassium chloride solution, and a flow rate of 0.8 ml min-1 was used throughout the investigation. The biosensor response was linear over the range of 0.075-30 mM glucose, with the former representing the detection limit. The precision of the system was determined by carrying out 20 repeat injections of a 5-mM glucose standard, and the calculated coefficient of variation was 3.9%. It was demonstrated that this biosensor system could be applied to the direct measurement of glucose in serum without pretreatment. Therefore, this would allow high-throughput analysis, at low cost, for this clinically important analyte. © 2009 Elsevier Inc. All rights reserved

Selective and rapid biosensor integrated into a commercial hand-held instrument for the measurement of ammonium ion in sewage effluent

A disposable biosensor for ammonium ions in sewage effluent was developed and based on a screen-printed carbon electrode coated with the enzyme glutamate dehydrogenase, 2-oxoglutarate, and NADH. This rapid and selective amperometric biosensor is capable of detecting NH4+ in sewage effluent at concentrations in the range 1-10 ppm. The biosensor was developed as a disposable, reagentless device incorporated into a fully automated, hand-held instrument that can be used away from the laboratory. This low cost, commercially available system permits the measurement of NH4+ in effluent samples in only 5 min and 20 s without any pretreatment. The results obtained correlated well with the standard laboratory method, used by the Environment Agency. Copyright © Taylor & Francis Group, LLC

Application of screen-printed microband biosensors to end-point measurements of glucose and cell numbers in HepG2 cell culture

Microband glucose biosensors were produced by insulating and sectioning through a screen-printed, water-based carbon electrode containing cobalt phthalocyanine redox mediator and glucose oxidase enzyme. Under quiescent conditions at 37 °C, at an operating potential of +0.4 V, they produced an amperometric response to glucose in buffer solutions with a sensitivity of 26.4 nA/mM and a linear range of 0.45 to 9.0 mM. An optimal pH value of 8.5 was obtained under these conditions, and a value for activation energy of 40.55 kJ mol-1 was calculated. In culture medium (pH 7.3), a sensitivity of 13 nA/mM was obtained and the response was linear up to 5 mM with a detection limit of 0.5 mM. The working concentration was up to 20 mM glucose with a precision of 11.3% for replicate biosensors (n = 4). The microband biosensors were applied to determine end-point glucose concentrations in culture medium by monitoring steady-state current responses 400 s after transfer of the biosensors into different sample solutions. In conjunction with cultures of HepG2 (human Caucasian hepatocyte carcinoma) cells, current responses obtained in 24-h supernatants showed an inverse correlation (R2 = 0.98) with cell number, indicating that the biosensors were applicable for monitoring glucose metabolism by cells and of quantifying cell number. Glucose concentrations determined using the biosensor assay were in good agreement, for concentrations up to 20 mM, with those determined spectrophotometrically (R2 = 0.99). This method of end-point glucose determination was used to provide an estimated rate of glucose uptake for HepG2 cells of 7.9 nmol/(106 cells min) based on a 24-h period in culture. © 2008 Elsevier Inc. All rights reserved

Application of high temperature electronics packaging technology to signal conditioning and processing circuits

The requirement to install electronic power and control systems in high temperature environments has posed a challenge to the traditional limit of 125°C for high temperature exposure of electronics systems. The leap in operating temperature to above 200°C in combination with high pressures, vibrations and potentially corrosive environments means that different semiconductors, passives, circuit boards and assembly processes will be needed to fulfil the target performance specifications. Bare die mounted onto ceramic and insulated metal substrates can withstand higher temperatures than soldered surface mount devices on printed circuit boards. The results of the evaluation of electronic interconnect and substrate materials that have been submitted to temperatures of 250°C for up to 2000 hours will be presented, including details on novel adhesive formulations and high temperature insulated metal substrates. The materials and processes developed have been applied to the manufacture of high temperature circuits representative of analogue signal conditioning and processing, using silicon on insulator devices and passive components mounted into HTCC packages and onto thick film on ceramic substrates. Results of the characterisation of these devices and circuits at temperatures of 250°C for up to 2000 hours will be presented. This work forms part of the UPTEMP project has been set-up with support from UK Technology Strategy Board and the EPSRC, which started in March 2007 with 3 years duration. The project brings together a consortium of end-users (Sondex Wireline and Vibro-Meter UK), electronic module manufacturers (GE Aviation Systems Newmarket) and material suppliers (Gwent Electronic Materials and Thermastrate Ltd) with Oxford University - Materials Department, the leading UK high temperature electronics research centre

Development of high temperature electronics packaging technology for long term operation at 250°C

The requirement to install electronic power and control systems in high temperature environments has posed a challenge to the traditional limit of 125°C for high temperature exposure of electronics systems. The leap in operating temperature to above 200°C in combination with high pressures, vibrations and potentially corrosive environments means that different semiconductors, passives, circuit boards and assembly processes will be needed to fulfil the target performance specifications. The fundamental change to enable this step change in packaging performance to be implemented is to switch from the traditional soldered surface mount or through-hole packaged devices assembled onto FR4 printed circuit board materials to bare die mounted onto ceramic, insulated metal or polyimide based substrates with selected materials that are intrinsically more capable of withstanding the high temperatures without degradation for prolonged durations. Results of the evaluation of electronic interconnect materials and substrate technologies that have been submitted to temperatures of 250°C for up to 2000 hours will be presented; including novel adhesive formulations and insulated metal substrates. Phenomena such as thermal migration and material deterioration due to high temperature exposure in air and inert atmospheres will be described, backed up by non-destructive scanning acoustic microscopy (SAM) of samples assessed throughout the long term testing programme. This work forms part of the UPTEMP project has been set-up with support from UK Technology Strategy Board and the EPSRC, which started in March 2007 with 3 years duration. The project brings together a consortium of end-users (Sondex Wireline and Vibro-Meter UK), electronic module manufacturers (GE Aviation Systems Newmarket) and material suppliers (Gwent Electronic Materials and Thermastrate Ltd) with Oxford University - Materials Department, the leading UK high temperature electronics research centre