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A simplified method for manufacturing glass-insulated metal microelectrodes.

A simplified method to manufacture durable, glass-insulated, tungsten microelectrodes with sufficient control of the final electrode impedance is described. This method requires only two instruments, an electrolytic etcher for wires and pipette puller, for manufacturing these electrodes. The manufacture of these electrodes involves 3 steps: (1) etching tungsten wire to sharpen the tip, (2) insulating the electrode by pulling a glass pipette over the sharpened tungsten wire and (3) assessing and adjusting the tip exposure and impedance of the electrode to meet recording requirements. Control over the electrode impedance is easily accomplished by varying the distance between the uppermost portion of the heating coil and the sharpened wire tip before a glass pipette is pulled over the wire tip. This distance determines the area of tip exposure and also the location where the glass insulation ends and the exposed electrode tip begins. A performance test of these electrodes in a chronically prepared monkey showed that they were strong enough to repeatedly penetrate thickened dura mater without significant changes in impedance and to isolate cortical neuronal activity after these multiple penetrations. Furthermore, the strength of these microelectrodes eliminated the need to remove reactive granular tissue from the dura overlying the recording site.

Animals↗

Determination of extracellular bicarbonate and carbon dioxide concentrations in brain slices using carbonate and pH-selective microelectrodes.

The extracellular pH of the brain is subject to shifts during neural activity. To understand these pH changes, it is necessary to measure [H+], [HCO3-], [CO3(2-)] and [CO2]. In principle, this can be accomplished using CO3(2-) and pH-sensitive microelectrodes; however, interference from HCO3- and Cl-, and physiological changes in [HCO3-], complicate measurements with CO3(2-) electrodes. Calibration requires knowledge of slope response, interference constants and corrections for [HCO3-] shifts. We show that when [HCO3-] is altered at constant [CO2] in the absence of Cl-, the HCO3- interference cancels and the Nikolsky equation reduces to the Nernst equation for CO3(2-). Measurement of CO3(2-) slope response by this method yielded a value of 28.5 +/- 0.72 mV per decade change in [CO3(2-)]. In Cl(-)-containing solutions, interference coefficient for HCO3- and Cl- were determined by altering [HCO3-] at constant [CO2], changing [CO2] at constant [HCO3-], then solving the simultaneous Nikolsky equations for each transition. The mean interference constants corresponded to selectivity ratios of 245:1 and 1150:1 for CO3(2-) over HCO3- and Cl- respectively. To correct for possible changes in [HCO3-], the equilibrium relation between CO3(2-) and HCO3- was substituted into the Nikolsky equation to yield an equation in [CO3(2-)] and [H+]. By simultaneously measuring shifts in [H+] with a pH microelectrode, this equation is readily solved for [CO3(2-)]. These methods were tested by measuring [HCO3-] and [CO2] in experimental solutions, and in the extracellular fluid of rat hippocampal slices.

Animals↗

Laser exposure of Parylene-C insulated microelectrodes.

The polymer, Parylene-C, has proven to be a biocompatible insulation for microelectrodes. However, due to its inert nature, the removal of the insulation from the tips of microelectrodes is difficult. This paper describes the use of an ultraviolet laser system to micromachine Parylene-C insulation with photoablation to precisely expose an arbitrary shape recording or stimulating surface.

Lasers↗

Direct measurement of intracellular free magnesium in frog skeletal muscle using magnesium-selective microelectrodes.

Mg2+-selective microelectrodes have been used to measure the intracellular free Mg2+ concentration in frog skeletal muscle fibers. Glass capillaries with a tip diameter of less than 0.4 micron were backfilled with the Mg2+ sensor, ETH 1117. In the absence of interfering ions, they gave Nernstian responses between 1 and 10 mM free Mg2+. In the presence of an ionic environment resembling the myoplasm, the microelectrode response was sub Nernstian (18-24 mV) but still useful. The electrodes were calibrated before and after muscle-fiber impalements . In quiescent fibers from sartorius muscle (Rana pipiens), with resting membrane potentials not less than -82 mV, the intracellular free Mg2+ concentration was 3.8 +/- 0.41 (S.E.) mM (n = 58) at 22 degrees C. No significant change in the intracellular free Mg2+ was observed following extensive (approx. 6 h) incubation in Mg2+-free media. Increasing the external concentration of magnesium from 4 to 20 mM (approx. 15 min) produced a slow and small enhancement (1.8 mM) of [Mg2+]i, which was fully reverted when the divalent cation was removed from the bathing solution. No change in ionic magnesium resting concentration was observed when the muscle fibers were treated either with caffeine 3 mM or with Na+-free solutions. In depolarized muscle fibers (-23 +/- 2.7 mV) treated with 100 mM K+, the myoplasmic [Mg2+] was 3.7 +/- 0.45 (S.E.) mM, n = 6, immediately after the spontaneous relaxation of the contracture. Similar determinations in muscle fibers during stimulation at low frequency (5 Hz), and after fatigue development, showed no changes in the concentration of free cytosolic Mg2+. These results point out that [Mg2+]i is not modified under these three different experimental conditions.

Animals↗

Multibarreled glass-coated tungsten microelectrode for both neuronal activity recording and iontophoresis in monkeys.

A multibarreled glass-coated tungsten microelectrode suitable for single-unit recording and iontophoresis in chronically behaving animals is described. The microelectrode was stiff enough to pass through the dura matter of behaving monkeys, and can be applied as a microiontophoretic electrode. The electrode is easy to build and usable in stabilized conditions after repeated penetrations.

Animals↗

Miniature manipulator with microelectrode for chronic recording of single units from freely-moving animals.

We report a simple recording and stimulating system, a miniature manipulator with a microelectrode (MMM) that can be mounted on animal's skull. The system consists of a miniature manipulator and a microelectrode which are used in one body. Several MMMs can be implanted simultaneously into the brain as the part of the manipulator is small. With this device, recording of single units from several different cortices is possible from awake and unrestricted animals.

Animals↗

Histamine selective microelectrode based on a synthetic organic liquid ion exchanger.

A two-barrel organic ion-sensitive microelectrode has been developed to electrochemically detect histamine activity. The synthesis of the histamine-sensitive liquid ion exchanger, composed of tetrafluorophenylboron histamine plus 3-nitro-o-xylol, and the construction of the two-barrel microelectrode (0.1-0.5 micron tip diameter) sensitive to histamine based on such an exchanger is reported. The calibration curves, their slope, selectivity, stability and detection limits for different solutions are described. High selectivity for Na+, K+, Ca2+ was observed.

Biosensing Techniques↗

Localizing the site of generation of uni-quantal endplate potentials using two intracellular microelectrodes.

By recording with a microelectrode at either end of the endplate, the site of generation of uni-quantal endplate potentials (EPP) can be estimated. One method assumes that the logarithm of the amplitude of the signal declines linearly with distance along the cable [4, 8]. The uncertainties in this method are discussed. We show that even on a muscle cable the logarithm of the time integral of the EPP declines linearly with distance, falling to 1/e at a distance equal to the length constant, lambda. We conclude that the integral approach provides a theoretically sound method for localizing the sites of uniquantal EPP generation with two microelectrodes.

Animals↗

Valinomycin-based K+ selective microelectrodes with low electrical membrane resistance.

A valinomycin-based membrane phase for microelectrodes with relatively low electrical membrane resistances is described. Microelectrodes with tip diameters of about 1 micron exhibit resistances of about 10(10) omega. Extremely high K+ selectivities are obtained, e.g. a rejection of Na+ by a factor of 5000 and of acetylcholine by a factor of 3400. At a constant background of 140 and 500 mM Na+, the detection limit of the K+ sensor is at 1.6 X 10(-5) and at 2.5 X 10(-5) M K+, respectively.

Electric Conductivity↗

A motor-driven miniature multiple microelectrode manipulator for recording from the cortical neuropile of the awake monkey in operant behavior paradigms.

An improved version of the Blum and Feldman multiple microelectrode manipulator is described. The main innovation is a design based on miniaturization and light weight (75 g), which allows head mounting with minimal disturbance to the animal in awake monkey operant behavior studies. The major features of the previously described model are preserved: simultaneous independent manipulation of four microelectrodes, in order to record from at least four single units, that may be crosscorrelated. With the new design better adjustment of intermicroelectrode distances for studies of interneural distances of 300 microns to 7 mm, and an easier automatic, computer-controlled monitoring of distances transversed into the brain are possible.

Animals↗

Membrane potential measurement in isolated outer hair cells of the guinea pig cochlea using conventional microelectrodes.

Membrane potential of the isolated outer hair cells (OHCs) from the guinea pig cochlea was measured using conventional microelectrodes filled with 200 mM KCl. The resting membrane potential during superfusion with the standard physiological saline solution containing 3.5 mM K+ was -47.3 +/- 1.4 mV (N = 72), which was higher than those previously reported for isolated OHCs studied by using microelectrodes. Addition of ouabain (10(-5)-10(-3) M), the specific Na+, K+ ATPase inhibitor, depolarized the cell slowly and progressively, indicating the presence of low but definite Na+, K+ ATPase activity in the plasma membrane of OHCs. The magnitude of membrane potential was mainly dependent on the extracellular K+ concentration ([K+]O). A ten-fold increase of [K+]O depolarized the membrane potential by 49.6 +/- 1.0 mV (N = 58). A decrease of [Na+]O to one tenth of the control hyperpolarized the membrane potential by about 2 mV. Decreasing extracellular Cl- from 131.3 mM to 27.5 mM did not cause a significant change in the membrane potential. Using the Goldman-Hodgkin-Katz equation, assuming a negligible contribution of Cl- to the membrane potential and total monovalent cat ion concentration of the cytosol similar to the extracellular fluid, we calculated the permeability ratio of K+ versus Na+ to 131 +/- 19 and intracellular K+ concentration to 33.3 +/- 1.9 mM.

Animals↗

Stimulation with chronically implanted microelectrodes in the cochlear nucleus of the cat: histologic and physiologic effects.

The effects of several hours of continuous electrical stimulation in the cats' cochlear nucleus with chronically implanted activated iridium microelectrodes was investigated from the changes in the evoked response near the inferior colliculus and also by histologic evaluation of the stimulated tissue. The stimulating microelectrodes had geometric surface areas of 75-500 microns2. They were pulsed continuously for 4 h, at a pulse repetition rate of 200 Hz, using charge-balanced pulse pairs. The charge per phase was 1.8 or 3.6 nC/ph. The animals were sacrificed for histologic evaluation 2 h, or several days later. The only remarkable histologic change resulting from the 4 h of stimulation was some aggregation of lymphocytes at the site of stimulation. However, depression of the electrical excitability of neurons near the sites often persisted for several days after 4 h of stimulation at 3.6 nC/phase. The charge per phase of the stimulus pulse pair was correlated strongly with the depression of excitability, and there was a weaker correlation between the depression and the amplitude of the first phase of voltage transient induced across the electrode-tissue interface. The charge density, calculated from the geometric surface area of the stimulating electrodes, was poorly correlated with the severity of the depression. The findings suggest a means of detecting impending stimulation-induced neural damage while it is still reversible.

Animals↗

Sonochemically fabricated enzyme microelectrode arrays for the environmental monitoring of pesticides.

This paper describes the development of a novel sonochemically fabricated microelectrode based acetylcholinesterase and polyaniline carbon/cobalt phthalocyanine biosensor for the ultra-sensitive determination of pesticides. Arrays of this type are fabricated using microelectrode templates with population densities of 2 x 10(5) cm(-2). The enzymatic response of the sensors is inhibited upon incubation with the pesticide and in this report it is shown that paraoxon may be determined down to concentrations of 1 x 10(-17) M. This sensitivity has thus far not been achieved and mechanisms accounting for the enhancement of the sensitivity reported here are discussed.

Acetylcholine↗

Thin-film IrOx pH microelectrode for microfluidic-based microsystems.

Microsensors are valuable tools to monitor cell metabolism in cell culture volumes. The present research describes the fabrication and characterization of on-chip thin-film iridium oxide pH microsensors with dimensions of 20 microm x 20 microm and 20 microm x 40 microm suitable to be incorporated into nl volumes. IrOx thin films were formed on platinum microelectrodes by electrochemical deposition in galvanostatic mode. Anodically grown iridium oxide films showed a near super-Nernstian response with a slope of -77.6+/-2 mV/pH at 22 degrees C, and linear responses within the pH range of 4-11. Freshly deposited electrodes showed response times as low as 6s. Long-term studies showed a baseline drift of 2-3 mV/month, which could easily be compensated by calibration. This work demonstrated for the first time the use of planar IrOx pH microelectrodes to measure the acidification rate of CHO and fibroblast cells in an on chip cell culture volume of 25 nl with microfluidic control.

Animals↗

Sonochemically fabricated microelectrode arrays for biosensors. Part III. AC impedimetric study of aerobic and anaerobic response of alcohol oxidase within polyaniline.

A sonochemically fabricated alcohol oxidase enzyme micro-electrode array is reported. Sensors of this type were fabricated by first depositing an insulating polydiaminobenzene film on supporting gold electrodes. Sonication and subsequent ablation exposed discrete areas of the underlying conducting electrode, which collectively act as a microelectrode array. Electropolymerisation of aniline has been used to generate in situ polyaniline containing entrapped alcohol oxidase. The physical and electrochemical properties of these films were studied and reported within this paper. The final composites were shown to behave with microelectrode performance characteristics for the detection of aqueous ethanol concentrations.

Aerobiosis↗

A disposable on-chip phosphate sensor with planar cobalt microelectrodes on polymer substrate.

Disposable microsensors on polymer substrates consisting of fully integrated on-chip planar cobalt (Co) microelectrodes, Ag/AgCl reference electrodes, and microfluidic channels have been designed, fabricated, and characterized for phosphate concentration measurement in aqueous solution. The planar Co microelectrode shows phosphate-selective potential response over the range from 10(-5) to 10(-2)M in acidic medium (pH 5.0) for both inorganic (KH(2)PO(4)) and organic (adenosine 5'-triphosphate (ATP) and adenosine 5'-diphosphates (ADP)) phosphate compounds. This microfabricated sensor also demonstrates significant reproducibility with a small repeated sensing deviation (i.e. relative standard deviation (R.S.D.)<1%) on a single chip and a small chip-to-chip deviation (i.e. R.S.D.<2.5%). Specifically, while keeping the high selectivity, sensitivity, and stability of a conventional bulk Co-wire electrode, the proposed phosphate sensor yields advantages such as ease of use, cost effectiveness, reduced analyte consumption, and ease of integrating into disposable polymer lab-on-a-chip devices. The capability to sense both inorganic and organic phosphate compounds makes this sensor applicable in diverse areas such as environmental monitoring, soil extract analysis, and clinical diagnostics.

Biosensing Techniques↗

Flexible polyimide microelectrode array for in vivo recordings and current source density analysis.

This work presents implantable, flexible polymer-based probes with embedded microelectrodes for acute and chronic neural recordings in vivo, as tested on rodents. Acute recordings using this array were done in mice under urethane anesthesia and compared to those made using silicon-based probes manufactured at the Center for Neural Communication Technology, University of Michigan. The two electrode arrays yielded similar results. Recordings with chronically implanted polymer-based electrodes were performed for 60 days post-surgically in awake, behaving rats. The microelectrodes were used to monitor local field potentials and capture laminar differences in function of cortex and hippocampus, and produced response waveforms of undiminished amplitude and signal-to-noise ratios 8 weeks after chronic implantation. The polymer-based electrodes could also be connected to a lesion current to mark specific locations in the tissue. Current source density (CSD) analysis from the recordings depicted a source - sink-composition. Tissue response was assessed 8 weeks after insertion by immunochemical labeling with glial fibrillary acidic protein (GFAP) to identify astrocytes, and histological analysis showed minimal tissue reaction to the implanted structures.

Animals↗

Cyclic chronopotentiometric determination of sugars at Au and Pt microelectrodes in flowing solutions.

The main advantage of the application of cyclic chronopotentiometry (CCP) in end-column CE detection arises from the fact that the detection parameters and the magnitude of the analytical signal are (in contrast with other electrochemical detection methods) independent of the ohmic polarization of the solution caused by the separation current at the detection end of the capillary. CCP was used to determine sugars on platinum and gold microelectrodes after separation by CE. The results obtained with a gold microelectrode were better. Subsequently this detection method was used for quantitative determination of sugars in honeys and for their authentication.

Carbohydrates↗