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Direct measurement of potassium leak from single 3 M KCl microelectrodes.

Potassium loss was measured from individual microelectrodes filled with 3 M KCl. The potassium content of a distilled-water droplet into which a microelectrode was inserted for a known time interval was analyzed by helium-glow photometry. The rate of potassium loss from single microelectrodes (18-97 M omega) was within the detectable limits of this method. The rate of potassium loss appeared to decrease with increasing input resistance (determined in 0.16 M NaCl) for electrodes having a low resistance (18-50 M omega). Higher resistance electrodes (greater than 50 M omega) appeared to achieve a uniform rate of potassium loss of 3.5 fmol X s-1. Estimates of the rate of potassium loss calculated from the rate of swelling of impaled cells were identical to those measured directly on the same group of electrodes. We conclude the electrolyte loss not only occurs into a distilled-water drop but also during intracellular recording. An alternative method for fabrication of microelectrodes was tested. Of these electrodes, the ones having an input resistance greater than 50 M omega lost potassium at rates below 1 fmol X s-1.

Animals↗

A new microelectrode method for simultaneous measurement of pH and PCO2.

A liquid-membrane pH-PCO2 microelectrode is described for the simultaneous measurement of pH and PCO2 in tissues and body fluids. The microelectrode is simple and easy to fabricate. It can be used to measure HCO3 concentration in solutions at chemical equilibrium. In the physiological range the microelectrode response is linear with nearly Nernstian slopes for PCO2 (61.2 +/- 2.0 mV/log10 PCO2) and pH (63.7 +/- 1.9 mV/pH units) (n = 14) at 37 degrees C. The PCO2 response is independent of the solutions' pH, anionic composition, and presence of serum proteins. In randomly micropunctured rat surface proximal tubules, pH averaged 6.80 +/- 0.04 and PCO2 averaged 57.7 +/- 4.6 mmHg (n = 22), whereas in the adjacent peritubular capillaries pH was higher (7.27 +/- 0.03) but PCO2 was not different (55.7 +/- 4.6 mmHg) (n = 22). Systemic arterial PCO2 was significantly lower compared with the renal cortex and averaged 37.4 +/- 2.4 mmHg (n = 14). Directly measured and pH-PCO2 microelectrode-derived HCO3 concentrations in systemic arterial blood, surface fluid bathing the kidney, and randomly micropunctured proximal tubules were approximately equal.

Animals↗

Single-unit pH-sensitive double-barreled microelectrodes for extracellular use.

The purpose of this study is to systematically describe the construction of pH-sensitive double-barreled microelectrodes for extracellular use. The most important advantages of these microelectrodes are as follows: the reference and the pH barrels are next to each other, and therefore the measured pH is not affected by asymmetric or slowly spreading direct current potential. The diameter of the tip of the microelectrodes is between 7 and 35 micron. These pH-sensitive microelectrodes are generally stable and Nernstian. They can be used repeatedly both in vivo and in vitro to measure tissue extracellular fluid pH. Some applications are described.

Animals↗

Magnitude of microelectrode refinement in pallidotomy and thalamotomy.

The relative accuracy of starting point algorithms in microelectrode-guided stereotactic pallidotomy and thalamotomy was evaluated using postoperative magnetic resonance imaging (MRI) data. Multiplanar reformations were performed to align postoperative MRI in anterior-posterior, dorsal-ventral and mediolateral planes. Three-dimensional distance and direction from the pallidal and thalamic stereotactic starting points to the respective radiofrequency lesions were measured. Similar magnitude of microelectrode refinement in pallidotomy and thalamotomy suggested similar accuracy of algorithms used to set the stereotactic starting point. Fewer microelectrode-recording tracts were required to identify optimal lesioning sites in thalamotomy compared to pallidotomy. Lesions were consistently localized anterior and superior to the starting point and a refined starting point algorithm may reduce the number of microelectrode recording tracts.

Adult↗

Free calcium in rat papillary muscle at contraction assessed with Ca-selective microelectrodes.

Direct measurements of free intracellular calcium (Ca)i are needed for an understanding of the regulation of contractility. An on-line measurement of (Ca)i with Ca-selective microelectrodes in intact muscle strips provides a suitable means of investigating this problem, although considerable methodologic difficulties exist. Measurements of (Ca)i concentrations during muscle contraction have been carried out by different methods such as Ca-binding techniques and aequorin luminescence, but remain unsatisfying, since they were not performed on intact muscle strips. The authors' measurements were carried out with Ca-selective microelectrodes on rat papillary muscles (stretched to optimal length in a perfusion bath of 1.54 mL at 30 degrees C). The impalement of electrodes was considered adequate when the heights of Ca-electrode potential and membrane potential remained constant for more than twenty minutes. For provoking contractile responses, the authors replaced the normal Tyrode solution by a caffeine-containing contracture solution (content in mM: 0 NaCl [choline], 4 CaCl2, 30 KCl, 25 caffeine, 1.05 MgCl2). Ca-selective microelectrodes were calibrated before and after each measurement and only those impalements were taken as adequate that showed identical calibration curves before and after the experiment. They measured the (Ca)i at 20%, 50%, and 80% of maximal contractile force and obtained (Ca)i concentrations of 1.1 +/- 0.3 microM (at 20%), 3.6 +/- 1.2 microM (at 50%), and 11.8 +/- 0.27 microM (at 80%) (n = 6, +/- SEM). These results represent the fist on-line measurements of the myocardial (Ca)i concentrations with Ca++-selective microelectrodes in intact muscle strips during various degrees of contraction.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Printed circuit microelectrodes and their application to honeybee brain.

The application of printed circuit technology to the production of a new type of multi-channel microelectrode is described. 2. Recordings have been obtained from the the protocerebrum of the honeybee Apis mellifera L. using printed circuit microelectrodes in both restrained and free-moving preparations. 3. These recordings are compared with those previously obtained from conventional probe microelectrodes and are found to have similar characteristics of transient voltage changes superimposed on an undifferentiated high frequency background. 4. A wide range of development possibilities for the printed circuit microelectrode are discussed.

Action Potentials↗

[Ion-selective microelectrodes for intracellular measurements].

Ion-selective electrodes have been used for many years to analyze ion activities in electrolyte solutions. In recent years, these electrodes have been miniaturized and applied for measurements in the cell interior. This article describes basic principles for both the fabrication and the intracellular application of ion-selective microelectrodes, particularly focusing on double barreled liquid ion-exchanger microelectrodes. The use of ion-selective microelectrodes allows continuous and real-time measurements of intracellular ion activities of a single cell in both multicellular and isolated cell preparations under various conditions, such as changes in ion composition of the extracellular bathing fluid and exposure to certain drugs. With double-barreled ion-selective microelectrodes, the transmembrane potential and intracellular ion activities can be measured simultaneously in the same cell. Although there are still some problems or limitations to the intracellular application of ion-selective electrodes, this technique is useful for determining the actual movement of intracellular ions, and thereby to elucidate cellular mechanisms of membrane transport and other physiological functions.

Action Potentials↗

Acute microelectrode array implantation into human neocortex: preliminary technique and histological considerations.

OBJECT: Researchers at The Center for Neural Interfaces at the University of Utah have designed and produced a silicon-based high-density microelectrode array that has been used successfully in mammalian models. The authors investigate the ability to transfer array insertion techniques to humans and examine the acute response of human cortical tissue to array implantation. METHODS: Six patients who were scheduled to undergo temporal lobectomy surgery were enrolled in an Institutional Review Board-approved protocol. Before the patients underwent lateral temporal cortical resection, one or two high-density microelectrode arrays were implanted in each individual by using a pneumatic insertion device. Cortical tissue was then excised and preserved in formalin. The specimens were sectioned and stained for histological examination. Pneumatic insertion of a microelectrode array into human cortex in the operating room was feasible. There were no clinical complications associated with implantation and no evidence of significant insertion-related hemorrhage. Tissue responses ranged from mild cortical deformity to small focal hemorrhages several millimeters below the electrode tines. Based on initial results, the insertion device was modified. A footplate that mechanically isolates a small area of cortex and a calibrated micromanipulator were added to improve the reproducibility of insertion. CONCLUSIONS: A high-density microelectrode array designed to function as a direct cortical interface device can be implanted into human cortical tissue without acute clinical complications. Further modifications to the insertion device and array design are ongoing and future work will assess the functional significance of the tissue reactions observed.

Electrodes, Implanted↗

[Stereotactic microelectrode-guided posteroventral pallidotomy for Parkinson's disease].

OBJECTIVE: To assess postoperative effects of microelectrode-guided posteroventral pallidotomy (PVP) for Parkinson's disease. METHODS: Intraoperative microelectrode recordings and microstimulation were used to explore the globus pallidus to performance of posteroventral pallidotomy in 48 patients with Parkinson's disease (47 unilateral and 1 bilateral). Assessment was made at baseline preoperatively and at 6 months intervals postoperatively, with unified Parkinson's disease rating scale (UPDRS). RESULTS: All patients were significantly improved on the limbs contralateral to the lesion side 6 - 34 months after operation (mean 24 months). The improvement was seen in the 'on' or 'off' state: UPDRS scores with patients on levodopa were improved by an average of 28.7%, while off medication scores showed reductions (47.6%) at 24 months. There were no deaths and no visual complications, but there were 4 patients (8.3%) of a delayed contralateral limbs dystonia after pallidotomy. CONCLUSIONS: The techniques of microelectrode recording and microstimulation indicate the location of the internal capsule and optic tract, which allow easy identification of these structure and facilitate PVP target in conjunction with radiofrequency microelectrode stimulation.

Adult↗

[Review on the progress of peripheral nervous microelectrode].

OBJECTIVE: To review the progress and application of peripheral nervous microelectrode. METHODS: The recent articles on peripheral nervous microelectrode were extensively reviewed. The classification, the progress of the peripheral nervous microelectrode and its utilizable prospect in the control of electronic prosthesis were summarized. RESULTS: The microelectrodes had favorable functions of selective stimulation and recording. It provided an information transmitting interface between the electric prosthesis and peripheral nerves. CONCLUSION: Peripheral nervous signal is a feasible signal source to control electronic prosthesis.

Artificial Limbs↗

In vivo tissue pO2 measurements in hamster skinfold by recessed pO2 microelectrodes and phosphorescence quenching are in agreement.

OBJECTIVE: Phosphorescence quenching has been used successfully to optically measure in vivo blood pO2 in the microvasculature. Optical measurements have also been made in some tissues, but it is not clear whether these results accurately reflect tissue pO2. METHODS: Recessed pO2 microelectrodes and the phosphorescence quenching technique were used simultaneously to measure in vivo tissue pO2 in hamster skinfold. The optical window for phosphorescence quenching was focused around the tips of microelectrodes that were positioned in tissue regions at least 100 microns from large microvessels. RESULTS: Mean tissue pO2 measured by recessed pO2 microelectrodes was 18.4 +/- 1.7 (SE) Torr, and mean tissue pO2 determined from the time course of phosphorescence decay was 18.8 +/- 2.0 Torr (no significant difference). The two tissue pO2 measurements agreed over a wide range, from 2 to 46 Torr (r = 0.93, 39 paired measurements from six sites in 3 animals). There was no systematic change in the microelectrode tissue pO2 during the period of light excitation used for the optical method. CONCLUSIONS: Under the conditions of our study, sufficient amounts of porphyrin dye leaked from the vasculature and diffused into tissue, allowing accurate measurements of tissue pO2 by the phosphorescence quenching technique. Furthermore, the optical method did not deplete significant amounts of O2 from tissue during light excitation.

Animals↗

Carbon composite microelectrodes: charge percolation and electroanalytical performance.

Microelectrodes based on two different epoxy-graphite composites (Araldite-M/HY5162 and Araldite-PY302-2/HY943) that are compatible with organic solvents have been developed and characterized. The variation in the bulk conductivity with graphite particle loading is described by percolation theory and indicates that the particles interact strongly with one another. The percolation threshold is 52% v/v loading of graphite, and this composite exhibits a bulk conductivity of 15 S m(-1). Microdisk electrodes of 25-microm diameter were produced by first etching a microcavity at the tip of a platinum microelectrode, which was then packed with a composite containing 60% v/v graphite so as to optimize both electrical conductivity and the electrode stability in acetonitrile and methanol solutions. Solution phase voltammetry of ferrocene is nearly ideal, and the responses are dominated by radial diffusion (slow scan rates) and semi-infinite linear diffusion (fast scan rates). The microelectrodes display high signal-to-noise ratios, good sensitivity, and low detection limits. The response times given by the product of the resistance, R, and capacitance, C, are 7.5 x 10(-4) and 1.4 x 10(-1) s for the Araldite M and PY302-2 composites, respectively. Although these response times are significantly slower than those associated with microelectrodes based on carbon fibers or metal wires, they are sufficient for time-resolved electroanalytical applications. The long response times arise from the large composite resistances, 3.1 x 10(11) and 8.3 x 10(11) Omega cm(-2) for Araldite M and PY302-2, respectively. Voltammetry of ferrocene in the absence of deliberately added supporting electrolyte is also reported. Significantly, indistinguishable slopes and intercepts for a calibration curve of peak current vs ferrocene concentration where 2 < [ferrocene] < 50 microM are obtained in the presence and absence of supporting electrolyte.

Journal Article↗

On-chip generated mercury microelectrode for heavy metal ion detection.

In this paper, the on-chip generated mercury microelectrode for heavy metal ion detection has been presented. A mercury droplet (approximately 150 microm in diameter) is on-chip generated to form a microelectrode through the mercury microfluidics control. The mercury microelectrode is used to electrochemically detect the heavy metal ions. The sample solutions with different concentration of heavy metal ions (Pb(2+) and Cd(2+)) have been successfully detected on the mercury droplet microelectrode using the square wave stripping voltammetry.

Journal Article↗

Underpotential deposition and anodic stripping voltammetry at mesoporous microelectrodes.

Using the technique of liquid crystal templating a series of high surface area mesoporous platinum microelectrodes was fabricated. The underpotential deposition of metal ions at such electrodes was found to be similar to that at conventional platinum electrodes. The phenomena of underpotential deposition, in combination with the intrinsic properties of mesoporous microelectrodes (i.e. a high surface area and efficient mass transport) was exploited for the purpose of anodic stripping voltammetry. In particular the underpotential deposition of Ag(+), Pb(2+) and Cu(2+) ions was investigated and it was found that mesoporous microelectrodes were able to quantify the concentration of ions in solution down to the ppb range. The overall behaviour of the mesoporous electrodes was found to be superior to that of conventional microelectrodes and the effects of interference by surfactants were minimal.

Journal Article↗

Measurement of the Cytoplasmic pH in Nitella translucens: Comparison of Values Obtained by Microelectrode and Weak Acid Methods.

A comparison has been made between the use of two types of pH microelectrode and the weak acid method for determining the cytoplasmic pH of Nitella translucens at an external pH of 6. There was good agreement between the value obtained with glass pH microelectrodes (7.54 +/- 0.15 se) and that obtained using the weak acid 5,5-dimethyloxazolidine-2,4-dione (7.42 +/- 0.07 se). Plastic-insulated antimony microelectrodes gave a significantly lower value (6.74 +/- 0.15 se) possibly due to disruption of the insulation by the cell wall. The addition of 1 mM NaN(3) rapidly reduced the pH recorded by the glass pH microelectrodes to about 5.3. A smaller change was observed using the weak acid method. The relevance of this observation to recent work on indoleacetic acid transport is discussed.

Journal Article↗

Microelectrode measurements of the activity distribution in nitrifying bacterial aggregates.

Microelectrodes for ammonium, oxygen, nitrate, and pH were used to study nitrifying aggregates grown in a fluidized-bed reactor. Local reactant fluxes and distribution of microbial activity could be determined from the microprofiles. The interfacial fluxes of the reactants closely reflected the stoichiometry of bacterial nitrification. Both ammonium consumption and nitrate production were localized in the outer shells, with a thickness of approximately 100 to 120 mum, of the aggregates. Under conditions in which ammonium and oxygen penetrated the whole aggregate, nitrification was restricted to this zone; oxygen was consumed in the central parts of the aggregates as well, probably because of oxidation of dead biomass. A sudden increase of the oxygen concentration to saturation (pure oxygen) was inhibitory to nitrification. The pH profiles showed acidification in the aggregates, but not to an inhibitory level. The distribution of activity was determined by the penetration depth of oxygen during aggregate development in the reactor. Mass transfer was significantly limited by the boundary layer surrounding the aggregates. Microelectrode measurements showed that the thickness of this layer was correlated with the diffusion coefficient of the species. Determination of the distribution of nitrifying activity required the use of ammonium or nitrate microelectrodes, whereas the use of oxygen microelectrodes alone would lead to erroneous results.

Journal Article↗

Five topographically organized fields in the somatosensory cortex of the flying fox: microelectrode maps, myeloarchitecture, and cortical modules.

Five somatosensory fields were defined in the grey-headed flying fox by using microelectrode mapping procedures. These fields are: the primary somatosensory area, SI or area 3b; a field caudal to area 3b, area 1/2; the second somatosensory area, SII; the parietal ventral area, PV; and the ventral somatosensory area, VS. A large number of closely spaced electrode penetrations recording multiunit activity revealed that each of these fields had a complete somatotopic representation. Microelectrode maps of somatosensory fields were related to architecture in cortex that had been flattened, cut parallel to the cortical surface, and stained for myelin. Receptive field size and some neural properties of individual fields were directly compared. Area 3b was the largest field identified and its topography was similar to that described in many other mammals. Neurons in 3b were highly responsive to cutaneous stimulation of peripheral body parts and had relatively small receptive fields. The myeloarchitecture revealed patches of dense myelination surrounded by thin zones of lightly myelinated cortex. Microelectrode recordings showed that myelin-dense and sparse zones in 3b were related to neurons that responded consistently or habituated to repetitive stimulation respectively. In cortex caudal to 3b, and protruding into 3b, a complete representation of the body surface adjacent to much of the caudal boundary of 3b was defined. Neurons in this area habituated rapidly to repetitive stimulation. We termed this caudal field area 1/2 because it had properties of both area 1 and area 2 of primates. In cortex caudolateral to 3b and lateral to area 1/2 (cortex traditionally defined as SII) we describe three separate representations of the body surface coextensive with distinct myeloarchitectonic appearances. The second somatosensory area, SII, shared a congruent border with 3b at the representation of the nose. In SII, the overall orientation of the body representation was erect. The lips were represented rostrolaterally, the digits were represented laterally, and the toes were caudolateral to the digits. The trunk was represented caudally and the head was represented medially. A second complete representation, PV, had an inverted body representation with respect to SII and bordered SII at the representation of the distal limbs. The proximal body parts were represented rostrolaterally in PV. Finally, caudal to both SII and PV, an additional representation, VS, shared a congruent border with the distal hindlimb representation of both SII and PV. VS had a crude topography, and receptive fields of neurons in VS were relatively large. Many neurons in VS responded to both somatosensory and auditory stimulation.

Animals↗

Fabrication of a gold microelectrode for amperometric detection on a polycarbonate electrophoresis chip by photodirected electroless plating.

A novel method of photoresist-free micropatterning coupled with electroless gold plating is described for the fabrication of an integrated gold electrode for electrochemical detection (ED) on a polycarbonate (PC) electrophoresis microchip. The microelectrode layout was photochemically patterned onto the surface of a PC plate by selective exposure of the surface coated without photoresist to 254 nm UV light through a chromium/quartz photomask. Thus, the PC plate was selectively sensitized by formation of reactive chemical moieties in the exposed areas. After a series of wet chemistry reactions, the UV-exposed area was activated with a layer of gold nanoparticles that served as a seed to catalyze the electroless plating. The gold microelectrode was then selectively plated onto the activated area by using an electroless gold plating bath. Nonselective gold deposition on the unwanted areas was eliminated by sonication of the activated PC plate in a KSCN solution before electroless plating, and the adhesion of the plated electrodes to the PC surface was strengthened with thermal annealing. Compared with the previously reported electroless plating technique for fabrication of microelectrodes on a microchip, the present method avoided the use of a membrane stencil with an electrode pattern to restrict the area to be wet-chemically sensitized. The CE with integrated ED (CE-ED) microchip was assembled by thermal bonding an electrode-plated PC cover plate to a microchannel-embossed PC substrate. The novel method allows one to fabricate low-cost, electrode-integrated, complete PC CE-ED chips with no need of a clean room. The fabricated CE-ED microchip was demonstrated for separation and detection of model analytes, including dopamine (DA) and catechol (CA). Detection limits of 0.65 and 1.03 microM were achieved for DA and CA, respectively, and theoretical plate number of 1.4 x 10(4) was obtained for DA. The plated gold electrode can be used for about 4 h, bearing usually more than 100 runs before complete failure.

Catechols↗