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[Microelectrode-guided posteroventral medial pallidotomy for Parkinson's disease].

OBJECTIVE: To introduce the methods and effects of microelectrode-guided posteroventral pallidotomy (PVP) for Parkinson's disease. METHODS: 109 patients underwent PVP by microelectrode electrophysiological recording for intraoperative target localization. 45 patients underwent unilateral PVP, 21 patients simultaneous bilateral PVP, 43 patients combined thalamotomy and PVP. Modified Webster Scale was used for objective assessments before and after operation. Postoperative CT scan or MRI was performed to localise lesions. RESULTS: Microelectrode recording usually led to a final pallidotomy lesion position that deviated from the CT stereotactically defined target point. The change rate of targets was 84.7%. PVP significantly and immediately improved all Parkinsonian motor signs and reduced drug-induced motor fluctuations and dyskinesia. The mean Webster Scale was improved by (72.7 +/- 11.3)% in the "on"" state, and (89.3 +/- 8.1)% in the "off" state. No patient showed permanent complications. CONCLUSIONS: Microelectrode-guided PVP was proved to be safe and effective. Bilateral PVP and combined PVP can significantly and immediately abolish all Parkinsonian motor signs. Physiological methods of microelectrode recording can significantly improve the safety and efficacy of PVP, and decrease the rate of complication.

Adult↗

[Microelectrodes and their application in diagnostic medicine].

Microelectrodes are in common use in medical detection systems. The binding of two complementary nucleic acid sequences is called hybridization. Today the major obstacle of large-scale hybridization approaches is the large time-dependency of a single reaction, which is up to 16 hours. As the DNA molecules can be electronically charged, the binding could be facilitated and confirmed using an electronic control system. The authors' team aimed to develop a microelectrode system capable for the detection and control of hybridization. A microelectrode head is immersed in small liquid drop. Here, the platinum counterelectrode is surrounded by a non-conducting quartz capillary. The reference electrode is chloridized silver immersed in saturated Ag/Cl dilution. The Ag/AgCl/1 M KCl +AgCl microelectrode in stabilized against the calomel electrode in the first hours, and remains stable between 7th and 30th hours. This can be verified by the minimal drop in the potential difference. Thus the AgCl saturated KCl electrode is usable for several days for actual measurements. The detector is controlled by an attached computer. The system can be used to detect hybridization in a micro-cell located on a gold-plate. The electrode can be dismounted and reused after repeated chloridization of the Ag wire. The microelectrode is simple, cheap; thus is best suited for application in future automated diagnostic detection systems.

Diagnostic Techniques and Procedures↗

The physical state of potassium in frog skeletal muscle studied by ion-sensitive microelectrodes and by electron microscopy: interpretation of seemingly incompatible results.

According to the commonly accepted membrane pump theory most of cellular K+ ions are freely dissolved in free cellular water; the alternative association-induction hypothesis postulates that the bulk of cellular K+ is adsorbed (weakly bound) to cellular proteins which are maintained in a specific labile state in the cytoplasm of a living cell. K+ activities measured with ion-sensitive microelectrodes in the cytoplasm of frog skeletal muscle seem to confirm the claim that most of cellular K+ ions are free in cellular water. On the other hand, it is evident from electron microscopic ion binding studies that in frog skeletal muscle most of cellular K+ ions are adsorbed to cellular proteins. The conflicting results can be explained with the assumption that a damage of the cytoplasm caused by the impaling microelectrode leads to a liberation of adsorbed ions. The possibility that microelectrodes damage the muscle cytoplasm is tested by using the light microscope. It is found that microelectrodes produce visible traumas which increase with time. Electron microscopic ion binding studies with damaged muscle support the view that monovalent cations are liberated in the disturbed area of a muscle fiber. It is concluded that a K(+)-sensitive microelectrode is not suited to determine the concentration of free K+ ions in intact frog skeletal muscle.

Animals↗

Ion-selective microelectrodes: theory and technique.

This report reviews the use of ion-selective microelectrodes to measure intracellular ionic activities and ionic electrochemical potential differences across cell membranes. Particular emphasis is placed on the electrochemical characteristics of liquid ion-exchanger microelectrodes. Methods for assessing the effect of interfering ions on the electrode potential are discussed and analyzed. An equivalent electrical circuit model is proposed in which deviations from their theoretical values of the slopes and selectivities of liquid ion-exchanger microelectrodes are analyzed in terms of surface conductance phenomena. A quantitative expression is developed that permits the transmembrane electrochemical potential difference for an ion, which is measured with an ion-selective microelectrode, to be corrected for deviations from ideality in the slope of the electrode response. The effect of lipophilic anions, dissolved in the organic ion-exchanger solution, on the electrochemical characteristics of cation selective liquid ion-exchanger microelectrodes containing neutral ionophores is discussed.

Animals↗

Voltammetric procedure for trace metal analysis in polluted natural waters using homemade bare gold-disk microelectrodes.

Voltammetric procedures for trace metals analysis in polluted natural waters using homemade bare gold-disk microelectrodes of 25- and 125-microm diameters have been determined. In filtered seawater samples, square wave anodic stripping voltammetry (SWASV) with a frequency of 25 Hz is applied for analysis, whereas in unfiltered contaminated river samples, differential pulse anodic stripping voltammetry (DPASV) gave more reliable results. The peak potentials of the determined trace metals are shifted to more positive values compared to mercury drop or mercury-coated electrodes, with Zn always displaying 2 peaks, and Pb and Cd inversing their positions. For a deposition step of 120 s at -1.1 V, without stirring, the 25-microm gold-disk microelectrode has a linear response for Cd, Cu, Mn, Pb and Zn from 0.2 microg L(-1) (1 microg L(-1) for Mn) to 20 microg L(-1) (30 microg L(-1) for Zn, Pb and 80 microg L(-1) for Mn). Under the same analytical conditions, the 125-microm gold-disk microelectrode shows linear behaviour for Cd, Cu, Pb and Zn from 1 microg L(-1) (5 microg L(-1) for Cd) to 100 microg L(-1) (200 microg L(-1) for Pb). The sensitivity of the 25-microm electrode varied for different analytes from 0.23 (+/-0.5%, Mn) to 4.83 (+/-0.9%, Pb) nA L micromol(-1), and sensitivity of the 125-microm electrode varied from 1.48 (+/-0.7%, Zn) to 58.53 (+/-1.1%, Pb nA L micromol(-1). These microelectrodes have been validated for natural sample analysis by use in an on-site system to monitor Cu, Pb and Zn labile concentrations in the Deûle River (France), polluted by industrial activities. First results obtained on sediment core issued from the same location have shown the ability of this type of microelectrode for in situ measurements of Pb and Mn concentrations in anoxic sediments.

Journal Article↗

Neuronal cell loss accompanies the brain tissue response to chronically implanted silicon microelectrode arrays.

Implantable silicon microelectrode array technology is a useful technique for obtaining high-density, high-spatial resolution sampling of neuronal activity within the brain and holds promise for a wide range of neuroprosthetic applications. One of the limitations of the current technology is inconsistent performance in long-term applications. Although the brain tissue response is believed to be a major cause of performance degradation, the precise mechanisms that lead to failure of recordings are unknown. We observed persistent ED1 immunoreactivity around implanted silicon microelectrode arrays implanted in adult rat cortex that was accompanied by a significant reduction in nerve fiber density and nerve cell bodies in the tissue immediately surrounding the implanted silicon microelectrode arrays. Persistent ED1 up-regulation and neuronal loss was not observed in microelectrode stab controls indicating that the phenotype did not result from the initial mechanical trauma of electrode implantation, but was associated with the foreign body response. In addition, we found that explanted electrodes were covered with ED1/MAC-1 immunoreactive cells and that the cells released MCP-1 and TNF-alpha under serum-free conditions in vitro. Our findings suggest a potential new mechanism for chronic recording failure that involves neuronal cell loss, which we speculate is caused by chronic inflammation at the microelectrode brain tissue interface.

Animals↗

Correlation of magnetic resonance and oxygen microelectrode measurements of carbogen-induced changes in tumor oxygenation.

PURPOSE: The aim of this work was to test the hypothesis that decreases in the linewidth of magnetic resonance (MR) water signals in tumors caused by oxygenating treatments are due to increases in capillary and venous oxygen saturation of hemoglobin, which are tightly coupled to increases in extravascular oxygen tension (pO2). To establish this link, changes measured by MR were compared to changes in tissue pO2 measured directly by oxygen microelectrodes during carbogen (95% O2/5% CO2) inhalation. METHODS AND MATERIALS: Mammary adenocarcinomas (R3230AC) in nine rats were imaged at 4.7 Tesla. T1-weighted (TR = 200 ms, flip angle = 45 degrees) spectroscopic images of the water resonance in a single slice through each tumor were acquired with spectral resolution of 3.9 Hz and bandwidth of +/-1000 Hz. In the same slices in these tumors, microelectrode measurements were made using a non-Clark style oxygen electrode with a 350-micron tip. MR and microelectrode measurements were made during alternating periods of air and carbogen inhalation. RESULTS: Water resonance linewidth decreased significantly during carbogen-induced hyperoxia. Paired Student's t-test analysis of microelectrode data indicated that pO2 was significantly (p < 0.05) increased as a result of carbogen inhalation. MR and microelectrode data averaged over each tumor demonstrated that decreased MR water signal linewidth is strongly correlated (r = 0.92, p < 0.05) with increased tumor pO2 levels. CONCLUSION: Although tumor oxygenating agents increase response to radiation in rodent tumors, clinical studies have shown only marginal effects on the radiosensitivity of human tumors. This may be, in part, because the effects of tumor oxygenating treatments are highly heterogeneous both within each tumor and among a population of tumors. The noninvasive, high-resolution MR methods that are validated by the present work could guide the design of new and more effective tumor oxygenating agents and optimize treatments for individual patients.

Animals↗

Hot microelectrodes.

Heat generation at disk microelectrodes by a high-amplitude (few volt) and high-frequency (0.1-2 GHz) alternating voltage is described. This method allows changing electrode temperature very rapidly and maintaining it well above the boiling point of solution for a very long time without any indication of boiling. The size of the hot zone in solution is determined by the radius of the electrode. There is no obvious limit in regard to the electrode size, so theoretically, by this method, it should be possible to create hot spots that are much smaller than those created with laser beams. That could lead to potential applications in medicine and biology. The heat-generating waveform does not electrically interfere with normal electroanalytical measurements. The noise level at hot microelectrodes is only slightly higher, as compared to normal microelectodes, but diffusion-controlled currents at hot microelectrodes may be up to 7 times higher, and an enhancement of kinetically controlled currents may be even larger. Hot microelectrodes can be used for end-column detection in capillary electrophoresis and for in-line or in vivo analyses. Temperature gradients at hot microelectrodes may exceed 1.5 x 10(5) K/cm, which makes them useful in studies of Soret diffusion and thermoelectric phenomena.

Journal Article↗

An independent, temperature-controllable microelectrode array.

Rapid, localized temperature control and negligible power consumption are key requisites for realizing effective parallel and sequential processing in the miniaturized, integrated biomedical microdevices where temperature-dependent biochemical reactions and fluid flow occur. In this study, an independent, temperature-controllable microelectrode array, with excellent temperature control rates and minimal power consumption, has been developed using microelectromechanical systems technology. The microfabricated array consists of Pt microelectrodes (100-microm diameter), with n-doped polysilicon microheaters (1.4-k Omega resistance), and vacuum-sealed cavities of depth 6.2 microm and diameter 200 microm. The thermal characteristics of each microelectrode were evaluated electrochemically through surface temperature measurements. The large heater power coefficient (2.1 +/- 0.1 degrees C mW(-1)) and the short heating and cooling times (less than 0.2 s for T(0.95)) are consequences of the vacuum-sealed cavities, which facilitate good thermal isolation and low thermal mass. The temperature of each microelectrode is independently controlled by a dedicated microheater, without thermally influencing the adjacent microelectrodes significantly.

Journal Article↗

Individually addressable recessed gold microelectrode arrays with monolayers of thio-cyclodextrin nanocavities.

Four, individually addressable 30 microm diameter, e-beam deposited, gold microelectrodes recessed by 6 microm were suitably spaced on a single substrate to avoid diffusional overlap between each microelectrode. The single substrate device was functionalised with thiolated alpha-, beta-, and gamma-cyclodextrin nanocavities without spacer groups to ensure close proximity of the cavities to the electrode surface. The microelectrodes were assessed in two stages. The e-beam deposited micron sized electrodes were characterized using models for recessed and inlaid microdisk electrodes. Subsequently, each individually addressable, atomically flat, micro-patterned gold electrode with thiolated CD ensembles was treated as a nanoporous electrode assembly. Theoretical and experimental results were compared using cyclic voltammetry. Atomic force microscopy was also used to characterise the modified microelectrodes. Comparisons were made with thiolated CDs deposited on macroelectrodes. This is the first report of the behaviour of immobilized CD nanocavities ensembles on atomically flat gold microelectrodes.

Journal Article↗

An improved method for constructing and selectively silanizing double-barreled, neutral liquid-carrier, ion-selective microelectrodes.

We describe an improved, efficient and reliable method for the vapour-phase silanization of multi-barreled, ion-selective microelectrodes of which the silanized barrel(s) are to be filled with neutral liquid ion-exchanger (LIX). The technique employs a metal manifold to exclusively and simultaneously deliver dimethyldichlorosilane to only the ion-selective barrels of several multi-barreled microelectrodes. Compared to previously published methods the technique requires fewer procedural steps, less handling of individual microelectrodes, improved reproducibility of silanization of the selected microelectrode barrels and employs standard borosilicate tubing rather than the less-conventional theta-type glass. The electrodes remain stable for up to 3 weeks after the silanization procedure. The efficacy of a double-barreled electrode containing a proton ionophore in the ion-selective barrel is demonstrated in situ in the leaf apoplasm of pea (Pisum) and sunflower (Helianthus). Individual leaves were penetrated to depth of approximately 150 microm through the abaxial surface. Microelectrode readings remained stable after multiple impalements without the need for a stabilizing PVC matrix.

Journal Article↗

[Ion-selective microelectrodes: principle and application of in vivo measurements of ionic concentrations in cochlear endolymph].

OBJECTIVE: To introduce the method of making neutral carrier ion-selective microelectrode and use it for in vivo measurement of ionic concentrations in cochloear endolymph. METHODS: GG-17 glass capillaries with 1.85 mm O.D. were rinsed extensively and dried in an oven. Two capillaries were parallelly connected and pulled with microelectrode puller to make a double-barreled micropipette. One barrel was back-filled with ion exchanger and internal reference solution, which served as ionic potential electrode, while the other barrel filled with 150 mmol/L KCL as reference electrode. Each barrel of microelectrode was connected to differential electrometer via Ag-AgCl wire and output was recorded on a three-channel recorder. Every ion-selective microelectrode was calibrated in a series of standard solutions to determine the required characteristics. Twenty healthy guinea pigs with normal hearing were anesthetized and were artificially respired through the tracheal canal after the intramuscular injection of suxamethonium chloride. The tympanic bulla was exposed and a double-barreled ion-selective electrode was inserted into the scala media through the round window and basilar membrane. The ionic potentials and endocochlear potentials (EP) were simultaneously recorded from the basal turn of the cochlea. Ionic concentrations were then calculated by Nicolsky-Eisenman equation. RESULTS: The concentrations of potassium, sodium and calcium ions in the cochlear endolymph were 146.3 +/- 11.8 mmol/L, 0.36 +/- 0.22 mmol/L and 16.2 +/- 5.7 micromol/L, respectively. CONCLUSIONS: The neutral carrier ion-selective microelectrode made with the above method is reliable and therefore can be used for accurately measuring the ionic concentrations of microenvironment in vivo continuously and transiently.

Animals↗

Use of microelectrodes to investigate the effects of 2-chlorophenol on microbial activities in biofilms.

In order to assess the applicability of using microelectrodes as a tool for inhibition tests, temporal and spatial inhibitory effects of 2-chlorophenol (2-CP) on O(2) respiration and nitrification activities in municipal wastewater biofilms were investigated using microelectrodes for O(2) and NH(4)(+). The time-course microelectrode measurements demonstrated that 2-CP inhibited O(2) respiration and nitrification activities within 6-18 min. The microbial activities were inhibited only in the upper 400 microm of the biofilms by 2-CP, and the bacteria present in the deeper parts of the biofilms were still active, probably due to limited penetration of 2-CP. These results could reasonably explain the difference in inhibitory ratios of the O(2) respiration and nitrification activities in the biofilms. O(2) respiration activity was incompletely inhibited, which was attributed to the presence of O(2) respiration activities in the deeper parts of the biofilm. In contrast, nitrification activity was significantly inhibited because ammonia-oxidizing bacteria were present in the upper parts of the biofilm. These results indicate that the microelectrodes with a very quick response time and a high spatial resolution are useful tools to study temporal and spatial inhibitory effects of inhibitors on in situ microbial activities in biofilms.

Bacterial Physiological Phenomena↗

A liquid ion-exchanger alternative to KCl for filling intracellular reference microelectrodes.

We have developed a filling solution for silanised microelectrodes consisting of potassium tetrakis (p-chlorophenyl) borate in octanol. Microelectrodes filled with this reference liquid ion-exchanger (RLIE) have equal selectivities to Na and K, and give the same membrane potential as do KCl-filled microelectrodes. The RLIE microelectrodes are more stable, less damaging to the cell membrane and do not leak Cl- ions. Their high resistance, however, makes them unsuitable for recording rapid potential changes or for passing current.

Animals↗

Intracellular neutral carrier-based Ca2+ microelectrode with subnanomolar detection limit.

In intracellular electrolyte solutions a Ca2+-selective microelectrode based on the synthetic electrically neutral carrier N,N,N',N'-tetracyclohexyl-3-oxapentanediamide (ETH 129) shows an improved detection limit when compared with the so far widely used Ca2+ microelectrodes based on the neutral carrier ETH 1001. Detection limits are found at pCa = 9.2 in Ca2+ buffers containing an intracellular background of K+ (125 mM). Selectivity studies in mixed solutions show a preference of Ca2+ over Na+ of 6 X 10(5), over K+ of 1.6 X 10(6), and over Mg2+ of 5 X 10(6). The microelectrode does not suffer from significant interference by inorganic and organic inhibitors and by lipophilic cations and anions. The low detection limit is unchanged at least during the first eight hours of continuous contact with Ca2+ solutions. The EMF drift during the first hour of use is between 5 and 10 mV and is then reduced to about 1 mV/h. The changes in EMF induced between solution of pCa = 7 and pCa = 8 are reproducible within 24.7 +/- 0.4 mV (SD, n = 8, about 3 h). These electrode characteristics were found for single-barrelled microelectrodes of one micrometer diameter front-filled with a PVC-containing membrane phase. In the absence of poly(vinyl chloride) in the membrane phase irregular EMF response curves were obtained throughout. Preliminary punctures of ferret ventricular muscle cells indicate that the Ca2+ electrode response is not disturbed by the contact of a cytosolic milieu.

Animals↗

The sensitivity of liquid sensor, ion-selective microelectrodes to changes in temperature and solution level.

Current procedures for the manufacture of ion-selective microelectrodes using liquid ion sensors result in two kinds of electrode, one with a short (10-100 micron) column of sensor at the tip and one with a long column (greater than 1 mm). We report here that the signal from a long-column microelectrode is very sensitive to small fluctuations in bath temperature and solution level when a recording is made above room temperature. This sensitivity is explained by the effects of temperature on the potentials developed at the interfaces between the ion sensor and the bordering aqueous solutions (the internal filling solution of the electrode and the test solution). If the temperature at the two interfaces is changed by different amounts, these interfacial potentials will also change by different amounts resulting in a change in the output of the electrode. Such an effect occurs for long-column electrodes because the column of sensor is usually not completely immersed in the heated test solution. Hence fluctuations in solution temperature and level will induce fluctuations in temperature along the column of sensor producing variations in the electrical signal from the electrode. In contrast, sensitivity to temperature and solution level is virtually absent in short-column microelectrodes since their column of sensor is fully immersed in the test solution and thereby at uniform temperature. In conclusion, in order to produce noise-free recordings of ion activities, the use of short-column microelectrodes is recommended for experiments performed above (or below) room temperature.

Electrophysiology↗

Piezoelectric translator. A simple and inexpensive device to move microelectrodes and micropipettes small distances rapidly.

A device is described that is capable of rapidly moving microelectrodes and micropipettes over distances up to 15 mu. This piezoelectric transLator uses the diaphragm from virtually any available piezoelectric buzzer in combination with simple physical support and drive electronics. All of the necessary details for the construction of this small device are presented. Each finished unit is about 2 cm long with a diameter of 2 cm and can be readily adapted to existing manipulators. The translator has been found useful in aiding the independent penetration by one or more microelectrodes of single cells or of more complicated multicellular preparations (including those that lie behind a connective tissue layer). This new device offers fine control of microelectrode motion that cannot be obtained by the other methods used to aid microelectrode and micropipette penetration of cell membranes (e.g. capacitance overcompensation--"ringing in"' or "tickling"--or tapping the manipulator base). Finally, the device described in this paper is extremely simple and inexpensive to build.

Cytological Techniques↗

Microelectrode study of K+ accumulation by tight epithelia: I. Baseline values of split frog skin and toad urinary bladder.

Toad bladder and split frog skin were impaled with fine-tipped single- and double-barrelled K+-selective microelectrodes. In order to circumvent membrane damage induced by impaling toad bladder, a null point method was developed, involving elevations of mucosal potassium concentration. The results suggest that intracellular potassium activity of short-circuited toad bladder is approximately 82 mM, twice as large as earlier estimates. Far more stable and rigorously defined intracellular measurements were recorded from short-circuited split frog skins. The intracellular positions of the micropipette and microelectrode tips were verified by transient hyperpolarizations of the membrane potential with mucosal amiloride or by transient depolarizations with serosal barium or strophanthidin. Simultaneous impalement of distant cells with separate micropipettes demonstrated that both the baseline membrane potentials and the responses to depolarizing agents were similar, further documenting that frog skin is a functional syncytium. Measurements with double-barrelled microelectrodes and simultaneous single-barrelled microelectrodes and reference micropipettes suggest that the intracellular potassium activity is about 104 mM, lower than previously reported. Taken together with measurements of intracellular potassium concentration, this datum suggests that potassium is uniformly distributed within the epithelial cells.

Animals↗