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At least 649 records · Page 36Linked to original sources
PubMed2003

Networks of neurons coupled to microelectrode arrays: a neuronal sensory system for pharmacological applications.

Two main features make microelectrode arrays (MEAs) a valuable tool for electrophysiological measurements under the perspective of pharmacological applications, namely: (i) they are non-invasive and permit, under appropriate conditions, to monitor the electrophysiological activity of neurons for a long period of time (i.e. from several hours up to months); (ii) they allow a multi-site recording (up to tens of channels). Thus, they should allow a high-throughput screening while reducing the need for animal experiments. In this paper, by taking advantages of these features, we analyze the changes in activity pattern induced by the treatment with specific substances, applied on dissociated neurons coming from the chick-embryo spinal cord. Following pioneering works by Gross and co-workers (see e.g. Gross and Kowalski, 1991. Neural Networks, Concepts, Application and Implementation, vol. 4. Prentice Hall, NJ, pp. 47-110; Gross et al., 1992. Sensors Actuators, 6, 1-8.), in this paper analysis of the drugs' effects (e.g. NBQX, CTZ, MK801) to the collective electrophysiological behavior of the neuronal network in terms of burst activity, will be presented. Data are simultaneously recorded from eight electrodes and besides variations induced by the drugs also the correlation between different channels (i.e. different area in the neural network) with respect to the chemical stimuli will be introduced (Bove et al., 1997. IEEE Trans. Biomed. Eng., 44, 964-977.). Cultured spinal neurons from the chick embryo were chosen as a neurobiological system for their relative simplicity and for their reproducible spontaneous electrophysiological behavior. It is well known that neuronal networks in the developing spinal cord are spontaneously active and that the presence of a significant and reproducible bursting activity is essential for the proper formation of muscles and joints (Chub and O'Donovan, 1998. J. Neurosci., 1, 294-306.). This fact, beside a natural variability among different biological preparations, allows a comparison also among different experimental session giving reliable results and envisaging a definition of a bioelectronic 'neuronal sensory system'.

Algorithms↗
PubMed2003

A portable microelectrode array recording system incorporating cultured neuronal networks for neurotoxin detection.

Cultured neuronal networks, which have the capacity to respond to a wide range of neuroactive compounds, have been suggested to be useful for both screening known analytes and unknown compounds for acute neuropharmacologic effects. Extracellular recording from cultured neuronal networks provides a means for extracting physiologically relevant activity, i.e. action potential firing, in a noninvasive manner conducive for long-term measurements. Previous work from our laboratory described prototype portable systems capable of high signal-to-noise extracellular recordings from cardiac myocytes. The present work describes a portable system tailored to monitoring neuronal extracellular potentials that readily incorporates standardized microelectrode arrays developed by and in use at the University of North Texas. This system utilizes low noise amplifier and filter boards, a two-stage thermal control system with integrated fluidics and a graphical user interface for data acquisition and control implemented on a personal computer. Wherever possible, off-the-shelf components have been utilized for system design and fabrication. During use with cultured neuronal networks, the system typically exhibits input referred noise levels of only 4-6 microVRMS, such that extracellular potentials exceeding 40 microV can be readily resolved. A flow rate of up to 1 ml/min was achieved while the cell recording chamber temperature was maintained within a range of 36-37 degrees C. To demonstrate the capability of this system to resolve small extracellular potentials, pharmacological experiments with cultured neuronal networks have been performed using ion channel blockers, tetrodotoxin and tityustoxin. The implications of the experiments for neurotoxin detection are discussed.

Action Potentials↗
PubMed1997

Hydrogen peroxide augmentation in a rat striatum after methamphetamine injection as monitored in vivo by a Pt-disk microelectrode.

We fabricated a Pt-disk microelectrode (diameter 30 microns) to conduct differential double-pulse amperometry (first step: 750 mV, 1 s; second step: 1,100 mV, 1 s) to detect hydrogen peroxide in the brain of a freely moving animal. This measurement determined hydrogen peroxide (detection limit, 0.03 microM) without any observable influence from other oxidizable species, such as dopamine (DA), ascorbic acid, or uric acid. The electrode was implanted into the right striatum of a rat. After intraperitoneal injection of methamphetamine (MAP), hydrogen peroxide concentrations were directly detected while the behavioral changes were monitored. MAP injection led to significant augmentation of hydrogen peroxide, the elevation of which depended on the dose of MAP. This is consistent with a previous report on the increase of DA-release caused by amphetamines and indirect evidence of the production of hydrogen peroxide via DA-metabolism.

Animals↗
PubMed2001

Microelectrode recording and macrostimulation in thalamic and subthalamic MRI guided stereotactic surgery.

Stereotactic neurosurgery aims at placing therapeutic lesions or chronic stimulating electrodes at very precise locations within the brain. Microelectrode recording and macrostimulation are used in addition to anatomoradiological techniques to optimize targeting. Recently, the usefulness of electrophysiological procedures has been questioned. Based on more than 500 therapeutic stereotactic lesions in the last 10 years at the thalamic and subthalamic levels, we evaluate here retrospectively the utility of the two electrophysiological procedures. In two of the three stereotactic targets considered in this study, intraoperative electrophysiological confirmation is mandatory because of the target size with respect to interindividual anatomical variations and of the more or less close vicinity of eloquent structures.

Adolescent↗
PubMed2001

Measurement of Ca(2+) currents in intact slow skeletal muscle fibers of the frog by the three-microelectrode technique.

The recording of currents passing through calcium channels in intact skeletal muscle fibers presents several difficulties. However, use of the three-microelectrode voltage-clamp technique at the end of the fiber provided us with a good approximation of current values in such fibers. Using this technique, we were able to measure the calcium-channel current in slow skeletal muscle fibers of the frog (Rana pipiens) and to quantify the effects of denervation on this current.

Animals↗
PubMed2003

Long-term stimulation of mouse hippocampal slice culture on microelectrode array.

To understand mechanisms of information processing, development and degeneration of the central nervous system, simultaneous multisite recording and stimulation have become extremely helpful. We have further developed the innovative approach to record from intact neural networks using planar microelectrode arrays (MEAs) with 60 substrate-integrated nano-columnar electrodes. To allow for long-term stimulation, mouse hippocampal tissue slices were immobilized onto MEAs and permanently moved between the gas and medium phase in a specifically designed tilting incubator that made it possible to electrically contact up to 90 MEAs with 5400 electrodes. After 2-3 weeks in vitro, histochemical staining, the intracellular microinjection of the fluorescent dye Alexa and the recording of spontaneous activity revealed in vivo-like characteristics of the organotypically cultured tissue. The feasibility of long-term stimulation during culturing was demonstrated with a low frequency paradigm. 0.003 Hz stimulation over a 16 h period resulted in a significant decline of field potentials and population spikes in two identified hippocampal subregions. Control experiments revealed that this effect was not due to tissue detachment or to induced cell death. In summary, the novel technology promises to open a new avenue for analyzing regulatory interactions of neuronal activity, cell differentiation and gene expression during development and in diseases.

Action Potentials↗
PubMed2002

Surface modified microelectrodes for selective electroanalysis of metal ions in environmental components.

The surface modification of electrodes was achieved by the Langmuir-Blodgett technique. The monolayers of laponite clay and polythiophene were formed at the air-water interface and these films were then transferred onto carbon microelectrodes. The behaviour of both untreated and coated electrodes was tested by originally developed double-step voltcoulometry (DSVCM). The dependence of charge response on the concentration of Cu(2+) species was investigated. Straight calibration curves were obtained and enhanced sensitivity of coated electrodes was documented. It is shown that the accumulation of Cu ions into laponite clay was maintained even after transferring the electrode into a pure water. The characteristic features of the "memory effect" are discussed.

Aluminum Silicates↗
PubMed1982

Microelectrode studies of the tegument and sub-tegumental compartments of male Schistosoma mansoni: an analysis of electrophysiological properties.

Standard intracellular microelectrode techniques were used to determine the electrical properties of the tegument and sub-tegumental regions in male Schistosoma mansoni. Three distinct compartments of electrical potential were observed. The resting potentials recovered in these compartments of -45.9 +/- 2.5 mV (Eteg), -22.0 +/- 1.1 mV (E2) and -4.7 +/- 03 mV (E3) corroborate those previously reported by Fetterer, Pax & Bennett (1980) and Bricker, Pax & Bennett (1981). Input resistance was measured in each compartment and was found to be 4. 5 M omega (tegument), 9.2 M omega (E2) and 3.5 M omega (E3). Time-constants for the tegument, E2 and E3 were 0.24 +/- 0.01 msec, 0.25 +/- 0.01 msec and 0.13 +/- 0.01 msec, respectively. Multiple electrode experiments revealed that the tegument and E2 compartment are electrical syncytia with similar current-spreading capabilities. Low resistance pathways also appear to connect the tegument and E2 region, since electrotonic signals initiated in either of those compartments experience only a 15-25% reduction upon passing into the other. Injecting large (greater than 200 nA) depolarizing current pulses into the tegument or E2 compartment often resulted in the initiation of active membrane responses. These spikes were highly variable, ranging from 4 to 75 mV in magnitude (occasionally overshooting zero potential by as much as 25 mV) and from 10-40 msec in duration. The responses were not actively propagated along the parasite, and their decay over distance was approximately equal to that predicted on the basis of length constant values obtained from electronic signals. The addition of a non-diffusible solute to the recording medium resulted in a significant reduction in the current-spreading capacity of both the tegument and E2 compartment. Coupling ratios between the tegument and E2 compartment. Coupling ratios between the tegument and E2 compartment were decreased, and the input resistance for both compartments increased, while resting potentials remained constant. Active responses could not be evoked in schistosomes exposed to the hyperosmotic medium.

Animals↗
PubMed1993

An ion-sensitive microelectrode study on the effect of a high concentration of ivermectin on chloride balance in the somatic muscle bag cells of Ascaris suum.

Ivermectin has been shown to increase chloride conductances of invertebrate cells. On the muscle cells of the parasitic nematode Ascaris, ivermectin acts as both a GABA receptor antagonist and a chloride channel opener. In this study, ion-sensitive microelectrodes were used to investigate the effect of ivermectin on intracellular Cl- concentration of the somatic muscle bag cells of Ascaris suum. Incubation of muscle cells with ivermectin (10 microM in 1% dimethyl sulphoxide vehicle for 60 min) increased intracellular Cl- by 2.9 mM or 15% compared to controls (P < 0.01, n = 6).

Animals↗
PubMed1990

Multi-microelectrode investigation of monkey striate cortex: link between correlational and neuronal properties in the infragranular layers.

Recordings were taken from infragranular layers of area 17 of anesthetized monkeys with an array of 30 microelectrodes matching about one hypercolumn. From intracortical spike-train correlations, the novel neuronal property "delay scale position" related to retino-cortical delays, was derived. Relationships were established to the degree of spike isolation and to classical response properties. Direction selectivity, spike rate, spike-isolation quality, delay scale, and color selectivity could be linked to an underlying factor upon which the latter variables depend in a fixed way. Neurons with similar factors were characterized by non-delayed correlations. The link was more strict in layer VI than in layer V, and it was related to the parvo/magnocellular subdivision of the visual system.

Action Potentials↗
PubMed1991

Characterization of the chemical architecture of carbon-fiber microelectrodes. 1. Carboxylates.

A new method to characterize the chemical architecture of a carbon-fiber microelectrode surface is described. Derivatization of carboxyl groups on the carbon surface with a poly(oxyalkalene)diamine (Jeffamine ED-600), followed by biotinylation of the free amine, allowed the attachment of a fluorescein isothiocyanate (FITC) conjugate of ExtrAvidin. The fluorescence observed after excitation at 488 nm was imaged with a fluorescence microscope equipped with a CCD camera, yielding a spatial map of the distribution of modified carboxyl groups on the surface of the carbon fiber with 0.5-micron resolution. Colloidal gold particles (15 nm diameter) coated with ExtrAvidin were used in place of the FITC-ExtrAvidin, and the carbon-fiber surface was imaged with scanning electron microscopy on a submicron scale. This selective information regarding surface-bound functional groups (i.e. carboxylates) has proven invaluable toward the rational design of novel sensors based on surface-modified ultramicroelectrodes.

Biotin↗
PubMed2000

Laser interference pattern ablation of a carbon fiber microelectrode: biosensor signal enhancement after enzyme attachment.

Fluorescence microscopy was used to visualize the accumulated fluorescent product of the enzyme alkaline phosphatase to indicate where active covalently bound enzyme remained on the surface after application of a Nd: YAG laser interference pattern to a surface that was first globally derivatized with the covalently bound enzyme. The electrochemical kinetics of the same carbon fiber surface were examined through the electrogenerated chemiluminescence of Ru(bpy)(3)2+ to determine that electron-transfer sites were indeed segregated from the enzyme-binding sites. The enzyme-derivatized areas are determined to be separate and distinct from the areas of enhanced electron transfer. Two other enzymes, glucose oxidase and malic dehydrogenase, were then covalently bound to carbon fiber microelectrode surfaces in order to verify the change in detection limit of their respective cofactors, NADH or H2O2, under a variety of surface conditions. The S/N of an enzyme-modified electrode after laser interference pattern photoablation and electrocatalytic treatment is improved by more than 1 order of magnitude over that observed at an electrode that is globally enzyme modified.

Biosensing Techniques↗
PubMed1993

Characterization of the chemical architecture of carbon-fiber microelectrodes. 2. Correlation of carboxylate distribution with electron-transfer properties.

Correlation of the chemical architecture of the surface of 10-microns-diameter carbon-fiber microelectrodes (illustrated by the fluorescence intensity of FITC-labeled carboxylates) and the rate of electron transfer of the surface (illustrated by the intensity of the electrogenerated chemiluminescence of luminol) allows the development of quantitative relationships between the chemical structure of an electrode surface and its electron-transfer properties. A fluorescence microscope equipped with a Peltier-cooled charge-coupled device was used to image these electrode surfaces with submicron spatial resolution. The total fluorescence emission observed at electrochemically treated electrodes was higher than that of controls while the voltammetric behavior and integrated ECL intensity of luminol were very similar. Imaging spectroscopy with submicron spatial resolution was able to demonstrate the microscopic heterogeneity of these surfaces and to assess the effect of the production of carboxylates on the rate of electron transfer of luminol.

Carbon↗
PubMed1994

gamma-irradiation-induced grafting of poly (styrenesulfonate) to poly(tetrafluoroethylene) shielded microelectrodes.

A new method for the fabrication and polymer modification of microelectrodes is described. These electrodes are constructed by heat sealing the electroactive material in dual shrink/melt poly(tetrafluoroethylene) (PTFE) under vacuum. The PTFE shield may be activated to provide a support upon which polymers of interest may be grafted. gamma-Irradiation was used to graft polymerize styrene to the surface. The poly-(styrene) was subsequently sulfonated with chlorosulfonic acid to form poly(styrenesulfonate). Scanning electron microscopy and Raman microspectroscopy provide evidence that the poly-(styrenesulfonate) film has been formed and extends over the electrode material. Voltammetry indicates that hexaammine-ruthenium(III) cation is preconcentrated and stabilized via an association with the polymer film.

Cation Exchange Resins↗
PubMed1989

Measurement of nanomolar dopamine diffusion using low-noise perfluorinated ionomer coated carbon fiber microelectrodes and high-speed cyclic voltammetry.

Several improvements in the fabrication and use of carbon fiber voltammetric microelectrodes (CFVMs) are described. These procedures did not involve oxidative treatment, but resulted in sensitivities and selectivities approaching those of treated CFVMs, without the inherent slow response times associated with the latter electrodes. To accomplish this we reduced CFVM noise by (1) improving the adhesive seal between the 8 microns o.d. carbon fiber and the glass insulation using vacuum, (2) snapping rather than cutting or beveling the fiber to be flush with the glass, and (3) using a concentrated electrolyte solution to make electrical contact with the fiber. System noise was reduced by digital smoothing and signal averaging. Selectivity of the CFVMs for dopamine over ascorbate was enhanced to better than 2000:1 by coating with Naflon, a perfluorinated cation exchange polymer, using a low (+0.5 V vs Ag/AgCl) electroplating potential. This low voltage also prevented electrode surface oxidation. To demonstrate the performance of our CFVMs, we used them in conjunction with high-speed cyclic voltammetry to accurately measure the diffusion coefficient of iontophoretically released dopamine at concentrations as low as 35 nM over distances of less than 200 microns in agarose gel.

Diffusion↗
PubMed1990

Simulation of two-electron homogeneous electrocatalysis for steady-state voltammetry at hemispherical microelectrodes.

Expanded space grid digital simulation of second-order, two-electron homogeneous electrocatalysis was extended to slow scan voltammetry at hemispherical microelectrodes. Predictions of the simulations are examined for reversible and quasireversible heterogeneous charge transfer of catalyst for a range of homogeneous catalytic rate constants (k1) and electrode radii. Working curves of catalytic efficiency vs long k1 were generated assuming reacting species with equal diffusion coefficients. As electrode radii in the less than 10-microns range decrease, progressively larger homogeneous catalytic rates are needed to yield analytically significant amplification of limiting currents. Simulations using hemispherical radii of (2/pi)rd can be used to predict catalytic efficiencies for microdisk electrodes with radii rd. Simulated working curves were used to estimate a log k1 of 3.88 +/- 0.55 (M-1 s-1) for electron transfer from the anion radical of 9,10-diphenylanthracene to 4,4'-dibromobiphenyl from steady-state catalytic efficiencies obtained at carbon microdisk electrodes. This value was in good agreement with 3.90 +/- 0.16 M-1 s-1 found previously by cyclic voltammetry.

Catalysis↗
PubMed2004

Dual microelectrodes for distance control and detection of nitric oxide from endothelial cells by means of scanning electrochemical microscope.

Dual Pt disk microelectrodes consisting of a 10-microm distance sensor and a 50-microm nitric oxide sensor were prepared. The 50-microm electrode was modified with Ni(4-N-tetramethyl)pyridyl porphyrin enclosed in the polymer network of a negatively charged electrodeposition paint. This paint prevented the dissolution of the otherwise soluble porphyrin in the aqueous test medium due to charge interactions. It also denied negatively charged ions in the analyte solution access to the electrode surface by electrostatic repulsion, thereby preventing interference from anions such as nitrite, nitrate, and ascorbate. With the aid of a scanning electrochemical microscope, it was possible to use the distance sensor by recording the negative feedback effect on the reduction of molecular oxygen to "guide" the nitric oxide sensor to various known distances from a layer of adherently growing human umbilical vein endothelial cells for the detection of nitric oxide released from the cells upon stimulation with bradykinin. The use of the distance sensor made it possible to preserve the integrity of the adherently growing cells concomitantly with the modified electrode by preventing the deterioration of the modifying layer during the distance adjustment step.

Cells, Cultured↗