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At least 865 records · Page 48Linked to original sources

Determination of traces of hemoglobin by square wave stripping voltammetry at a silver microelectrode.

An electrochemical method was developed for the determination of traces of hemoglobin (Hb) by adsorption square wave voltammetric stripping at a bare silver microelectrode. Under optimum conditions the proposed method provided a linear response over the hemoglobin concentration range 5 to 100 nmol/L. The detection limit was 3 nmol/L. The relative standard deviation was 4.5% for 6 successive determinations at 50 nmol/L Hb. Other chemicals present in the sample did not interfere.

Journal Article↗

Simultaneous recording of local electrical activity, partial oxygen tension and temperature in the rat hippocampus with a chamber-type microelectrode. Effects of anaesthesia, ischemia and epilepsy.

A miniature multiple thin-film recording sensor was used to measure simultaneously the electrical activity, oxygen content and temperature of brain tissue. The chamber-type potential sensor was an Ag/AgCl electrode covered by an Si3N4 (silicon nitride) chamber. The chamber-type oxygen sensor consisted of an Au-Ag/AgCl two-electrode electrochemical cell embedded in an electrolyte-filled Si3N4 chamber. The temperature sensor was a thin-film germanium resistor. The different sensors were spaced 300 microns apart. Anaesthetics (pentobarbital, chloral hydrate, chlornembutal, halothane) were shown to depress electrical activity and to increase local oxygen tension in the hippocampus. Halothane, but not the other anaesthetics, also increased the current output of the oxygen sensor when tested in saline bath, indicating that the apparent increase in measured oxygen levels during halothane anaesthesia was partly due to an electrochemical effect of halothane on the oxygen sensors. The decrease of tissue oxygen consumption produced by the other anaesthetics is likely to be the result of metabolic depression. Cerebral ischemia, evoked by cauterization of the vertebral arteries and occlusion of the carotid arteries for 30 min, resulted in the disappearance of both spontaneous and evoked electrical activity in the hippocampus and a decrease of both local temperature and oxygen tension. There was a marked overshoot of the oxygen tension to above preocclusion level following the release of the carotid arteries. As soon as electrical activity returned, the oxygen tension fell again, often below the lowest level seen during the ischemic period. This secondary decrease of oxygen level could be reversed by administration of supplementary small doses of anaesthetic. The anaesthetic-induced increase in oxygen tension was accompanied by a marked decrease in electroencephalogram amplitude and frequency. During electrically induced seizures a decrease in hippocampal oxygen content occurred and was accompanied by an increase of local temperature. Since the rectal temperature was kept constant, the changes in temperature are likely to reflect changes in blood perfusion of the recorded area. These findings are in agreement with previous observations made with conventional electrodes. In addition, the miniature size of the chamber-type microelectrode assembly allows a correlated monitoring of parallel physiological changes with high spatial and temporal resolution during anaesthesia, ischemia and epilepsy.

Action Potentials↗

Anatomy and physiology of the gerbil cochlear nucleus: an improved surgical approach for microelectrode studies.

A new, improved surgical approach to the cochlear nucleus is developed in the gerbil. This new approach involves making a small hole in the lateral wall of the temporal bone located within the perimeter of the superior semicircular canal. Microelectrodes are passed through the intact parafloccular lobe of the cerebellum to the cochlear nucleus. One advantage of the new approach is that no removal of any CNS vasculature or neural tissue is necessary. Relations between the bulla, temporal bone and cochlear nucleus are presented in detail. The new approach is demonstrated by making single unit recordings from the cochlear nucleus and classifying response patterns as measured in PST histograms. All of the response types found in cat are found in the gerbil.

Animals↗

Metabolic response of AH13r rat tumours to cyclophosphamide as monitored by pO2 and pH semi-microelectrodes.

The composition of the microenvironment has an important influence on the cellular response to cytotoxic agents. Using pH and pO2 semi-microelectrodes, we have monitored metabolic changes in AH13r rat tumours as a function of time after subcurative chemotherapy. Prior to therapy, tumours contained large areas considered hypoxic (mean pO2 approximately 4 mmHg) and are characterised by a marked accumulation of acidic metabolites (mean pH 6.65). Administration of cyclophosphamide (40 mg/kg body weight) resulted in tumour regression to 15% of pretreatment volumes and a growth delay of 12 days. Concomitant with volume reduction, tumours became reoxygenated (mean pO2 approximately 7 mmHg), with maximum values being reached within 2-4 days, paralleled by a shift of pH to more alkaline values (0.17 U on average). These changes coincided with the development of subtotal necrosis. During early tumour regrowth, the pH and pO2 histograms returned to control values. These data corroborate and extend the results of previous studies in which noninvasive techniques had been applied for the monitoring of treatment-induced metabolic changes in malignant tumours in vivo. In addition, these results support the notion that the effectiveness of anticancer therapy might be improved by selecting and scheduling therapeutic agents in consideration of physiological changes caused by preceding courses of treatment.

Animals↗

Micro-profiles of activated sludge floc determined using microelectrodes.

The microbial activity within activated sludge floc is a key factor in the performance of the activated sludge process. In this study, the microenvironment of activated sludge flocs from two wastewater treatment plants (Mill Creek Wastewater Treatment Plant and Muddy Creek Wastewater Treatment Plant, with aeration tank influent CODs of 60-120 and 15-35 mg/L, respectively) were studied by using microelectrodes. Due to microbial oxygen utilization, the aerobic region in the activated sludge floc was limited to the surface layer (0.1-0.2mm) of the sludge aggregate at the Mill Creek plant. The presence of an anoxic zone inside the sludge floc under aerobic conditions was confirmed in this study. When the dissolved oxygen (DO) in the bulk liquid was higher than 4.0mg/L, the anoxic zone inside the activated sludge floc disappeared, which is helpful for biodegradation. At the Muddy Creek plant, with its lower wastewater pollutant concentrations, the redox potential and DO inside the sludge aggregates were higher than those at the Mill Creek plant. The contaminant concentration in the bulk wastewater correlates with the oxygen utilization rate, which directly influences the oxygen penetration inside the activated sludge floc, and results in redox potential changes within the floc. The measured microprofiles revealed the continuous decrease of nitrate concentration inside the activated sludge floc, even though significant nitrification was observed in the bulk wastewater. The oxygen consumption and nitrification rate analyses reveal that the increase of ammonia flux under aerobic conditions correlates with nitrification. Due to the metabolic mechanisms of the microorganisms in activated sludge floc, which varies from one treatment plant to another, the oxygen flux inside the sludge floc changes accordingly.

Ammonia↗

Improved electrochemical detection of biogenic amines in Aplysia using base-hydrolyzed cellulose-coated carbon fiber microelectrodes.

A major challenge with electrochemical techniques in vivo, using carbon-fiber microelectrodes, is to achieve sufficient sensitivity to detect the low concentrations of transmitters released by neurons. In particular, when an electrode is inserted into living tissue, its sensitivity is usually substantially decreased as a result of the degradation of the carbon surface by proteins. Here, we show that this decrease can be significantly attenuated by coating the electrode with cellulose acetate. The cellulose film offers a steric barrier that prevents macromolecules from diffusing to the carbon surface and its porosity can be progressively increased by controlled hydrolysis. We compared different cellulose-coated electrodes, either non-hydrolysed or hydrolyzed, in 0.08 N KOH for 10-30 min. We found that dopamine and serotonin detection was blocked by non-hydrolysed cellulose films, but that hydrolysis restored optimal detection similar to uncoated electrodes. Moreover, cellulose films (hydrolyzed for 20 min) significantly diminished electrode degradation in vivo and allowed reliable detection of fast concentration changes with <0.5 s delay, compared to uncoated electrodes. Finally, the sensitivity to endogenous 5-HT release in Aplysia central nervous system was more than doubled with these electrodes. We conclude that the optimal hydrolysis time of cellulose-coated electrodes is approximately 20 min with our protocol and carbon fiber electrodes prepared with this method offer improved sensitivity for the detection of biogenic amines.

Animals↗

A three-enzyme microelectrode sensor for detecting purine release from central nervous system.

As the purines, in particular adenosine, are important signaling agents in the nervous system we have devised a new biosensor for directly measuring their production in real time during physiological activity. Our amperometric adenosine biosensor is made by entrapping 3 enzymes (xanthine oxidase, purine nucleoside phosphorylase and adenosine deaminase) in a composite lactobionamide and amphiphillic polypyrrole matrix around a Pt microelectrode. The resulting sensors are small (25-100 microm diameter), fast responding (10-90% rise time, 2+/-0.23 s), sensitive (100-222 mA M(-1) cm(-2)) and stable (100% activity after 5 days). The sensor was used in vivo to demonstrate the spatial localization of release of adenosine from Xenopus embryo spinal cord during fictive swimming.

Animals↗

Microelectrode study of single cavitational bubbles induced by 500 kHz ultrasound.

Insight is gained into about the processes governing cavitational activity and acoustic streaming induced by high frequency (500 kHz) ultrasound by the use of microelectrodes with short time resolution electrochemical equipment to allow monitoring of the activity of single cavitating bubbles. Current transients are interpreted as showing the flux of solution towards the electrode surface due to microstreaming. In order to explain the current amplitude, a simplified model is produced. Important parameters such as bubble size and shape on the surface as well as the boundary layer thickness for microstreaming are taken into account. This model leads to the amplitude of the oscillations of the cavitating bubble. Introducing realistic bubble sizes, this amplitude is found to be in the order of 1 micron. The conclusions arising from this work allow a further interpretation of previous observations at millimeter scale electrodes.

Journal Article↗

Determination of mosquito bloodmeal pH in situ by ion-selective microelectrode measurement: implications for the regulation of malarial gametogenesis.

Malarial gametocytes circulate in the peripheral blood of the vertebrate host as developmentally arrested intra-erythrocytic cells, which only resume development into gametes when ingested into the bloodmeal of the female mosquito vector. The ensuing development encompasses sexual reproduction and mediates parasite transmission to the insect. In vitro the induction of gametogenesis requires a drop in temperature and either a pH increase from physiological blood pH (ca pH 7.4) to about pH 8.0, or the presence of a gametocyte-activating factor recently identified as xanthurenic acid (XA). However, it is unclear whether either the pH increase or XA act as natural triggers in the mosquito bloodmeal. We here use pH-sensitive microelectrodes to determine bloodmeal pH in intact mosquitoes. Measurements taken in the first 30 min after ingestion, when malarial gametogenesis is induced in vivo, revealed small pH increases from 7.40 (mouse blood) to 7.52 in Aedes aegypti and to 7.58 in Anophĕles stephensi. However, bloodmeal pH was clearly suboptimal if compared to values required to induce gametogenesis in vitro. Xanthurenic acid is shown to extend the pH-range of exflagellation in vitro in a dose-dependent manner to values that we have observed in the bloodmeal, suggesting that in vivo malarial gametogenesis could be further regulated by both these factors.

Aedes↗

Microscopic measurement of pH with iridium oxide microelectrodes

Microscopic pH electrodes were produced by deposition of hydrous iridium oxide onto carbon fiber microelectrodes. The electrodes exhibit two linear regions of potentiometric response between pH 2-6 and pH 6-12. The electrodes respond to pH changes within 50 ms, and an equilibrium value is reached within 30 s. By using these electrodes as probes in the scanning electrochemical microscope, dynamic pH changes occurring at or near a surface can be measured and pH maps of the surface can be generated. Vertical pH profiles and images of pH were obtained at substrates where electrochemical (oxidation and reduction of H2O2, hydrogen evolution) or enzymatic (glucose oxidase) reactions involving proton transfers occur.

Journal Article↗

All-diamond microelectrode array device.

We report the development of all-diamond microelectrochemical devices, namely, a microelectrode array (MEA), in which a periodic array structure with well-defined diameters, distance, and hexagonal unit cell pattern is micromachined using a combination of state-of-the-art microwave-induced plasma growth and laser ablation shaping techniques to prepare and coat a patterned boron-doped diamond (BDD) substrate with an intrinsic diamond insulating layer. The active BDD element can be tuned to between 10 and 50 microm in diameter with a 10 times diameter center-to-center distance between two adjacent conducting elements, which are exactly coplanar to the dielectric surroundings. This type of device should enable applications in harsh conditions such as high temperature, high pressure, and resistive media under dynamic flow regimes.

Journal Article↗

Effect of change in angle between microelectrode surface and jet direction in flow system on current response in solutions of different ionic strength.

The influence of stream orientation versus surface of microelectrode detector was examined in the range between vertical and parallel flow for various jet velocities and various levels of supporting electrolyte. The flow cell was equipped with a conical body Pt microdisk electrode, and the measurements involved voltammetry and chronoamperometry. Ferrocene, two its charged derivatives (sodium ferrocenylo sulfate and ferrocenylomethyltrimethylamino hexafluorophosphate) and sodium iodide were employed as the substrates in the experiments. The strongest convectional transport and the highest signal of the analytes was obtained for alpha = 60 degrees (alpha is the angle between the electrode surface and the stream direction). The measured current increased by up to 1.85 times versus the traditional setup, and therefore, this new geometry of the detector is analytically advantageous. The value of alpha corresponding to the highest signal tended to decrease to approximately 45 degrees in the absence of supporting electrolyte provided that either flow rate or analyte concentration was above a certain threshold value. The experiments indicated that the interplay of the convectional and migrational components in the analyte transport is different for the charge increase and the charge cancellation processes. These experimental facts were confirmed by digital simulation results.

Journal Article↗

Fabrication and characterization of nanostructured Pd hydride pH microelectrodes.

Novel pH microsensors were made by electrodepositing mesoporous Pd films onto Pt microdisks, electrochemically loading the films with hydrogen to form the alpha+beta Pd hydride phase, and then switching to the potentiometric mode to monitor pH. To create a nanostructure, the films were deposited within a molecular template formed by the self-assembly of surfactant molecules, a technique known as true liquid crystal templating. The films retain the micrometer size of the Pt microdisk but offer electroactive areas up to 900 times larger. Optimum hydrogen loading conditions were determined, and the mesoporous Pd microdisks were found to have excellent potentiometric properties. From pH 2 to 12, their potential was Nernstian, highly reproducible, very stable (+/-1.2 mV over 2 h), and without hysteresis. Their response time was better than 1 s. However, the presence of oxygen reduced their lifetime significantly, thereby requiring frequent reloading. These microelectrodes do not require calibration before and after measurements, a procedure normally essential for potentiometric pH microsensors. To our knowledge, these are the first results where nanostructured materials made by the true liquid crystal templating method have been used in the potentiometric mode; moreover, these are the first results demonstrating the application of nanostructured microdisks in the potentiometric mode.

Journal Article↗

Voltammetric and reference microelectrodes with integrated microchannels for flow through microvoltammetry. 1. The microcell

This paper describes the construction of 2 microsensor units for on-line voltammetric detection inside a cylindrical microcell (a working microsensor unit and a reference and auxiliary microsensor unit), for application to heavy-metal analysis in complex media such as natural waters. Both microsensor units include a channel for the solution renewal in the microcell after analysis. The working microsensor, a Hg-plated Ir microelectrode, is protected against fouling with an agarose gel including a hydrophobic chromatographic phase (C18). The fabrication steps and the quality tests related to long-term use and reliability, as well as to precision, are described. The application of the protective gel layer against fouling by hydrophobic or surface active small molecules is of general application, reliable, and very efficient. The reference and auxiliary unit is composed by an iridium oxide based mini reference electrode, an auxiliary Pt electrode, and a circulation channel. It is built to enable its use inside a 700-microm-diameter tubing connected to a hollow fiber supported liquid membrane lumen (volume, 5-10 microL) for heavy-metal analysis. However, it can be used in any other microanalytical system. The reference electrode is sufficiently stable for voltammetric applications (1-2 mV drift/day), and its lifetime is more than one year. The Ti/IrO2 core is immersed in a pH-buffered agarose gel, to guarantee potential stability even when the electrode is immersed in variable pH solutions.

Journal Article↗

Subnanoliter volume wall-jet cells combined with interdigitated microarray electrode and enzyme modified planar microelectrode

Miniaturized wall-jet type flow cells with an active volume of 0.042-15 nL were fabricated for use as highly sensitive electrochemical detectors for capillary electrophoresis/electrochemical detection and small on-line enzyme sensors. The cells consisted of three glass plates and a fused-silica capillary. Two of the plates had microfabricated flow channels and guide trenches for the capillary and working, reference, and counter electrodes. The other plate had a film electrode. When an interdigitated microarray electrode (total area, 66 microm x 64 microm; bandwidth and gap, 2 microm) was installed in the flow cell, the redox cycling enhanced the current at flow rates of less than 100 nL/min even though there were only eight pairs of microbands. A sharp dopamine peak enhanced by the redox cycling was observed when the cell was used for capillary electrophoresis. A square film electrode modified with glutamate oxidase and Os-poly(vinylpyridine) containing HRP was also installed in the flow cell and used to measure neurotransmitter release from cultured nerve cells. When the flow rate was relatively high, the response time of the modified electrode was comparable to that of a cylindrical carbon fiber electrode (33 microm o.d.) modified with the same enzyme and mediator. We observed a transient cathodic current response assigned to the glutamate release with the electrode in the flow cell in a suction mode measurement when we stimulated cultured nerve cells electrically with a dual microelectrode.

Journal Article↗

In situ control of cellular growth and migration on substrates using microelectrodes.

We describe an electrochemical method to direct the growth and migration of mammalian cells on a substrate during cultivation in situ. Exposing the albumin-coated substrate to an oxidizing agent, hypobromous acid, electrochemically generated at the tip of the scanning microelectrode, locally switched the substrate from cytophobic to cell-adhesive. This transformation generated the formation of cellular micropatterns. Since the concentration of the oxidizing agent required for the surface processing did not cause significant damage to the cell cultures, we were able to direct in situ cellular proliferation and migration by drawing adhesive micropatterns over the preexisting cellular pattern.

Journal Article↗

Potentiometric biosensing of proteins with ultrasensitive ion-selective microelectrodes and nanoparticle labels.

We report here for the first time on the use of potentiometry for ultrasensitive nanoparticle-based detection of protein interactions. A silver ion-selective microelectrode is used to detect silver ions oxidatively released from silver enlarged gold nanoparticle labels in a sandwich immunoassay. Since potentiometry is expected to largely maintain its analytical characteristics upon reducing the sample volume, it is anticipated that this approach may form the basis for bioassays with attractive detection limits.

Biosensing Techniques↗