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PubMed1987

Fluxes of h and k in corn roots : characterization and stoichiometries using ion-selective microelectrodes.

We report here on an experimental system that utilizes ion-selective microelectrodes to measure the electrochemical potential gradients for H(+) and K(+) ions within the unstirred layer near the root surface of both intact 4-day-old corn seedlings and corn root segments. Analysis of the steady state H(+) and K(+) electrochemical potential gradients provided a simultaneous measure of the fluxes crossing a localized region of the root surface. Net K(+) influx values obtained by this method were compared with unidirectional K(+) ((86)Rb(+)) influx kinetic data; at any particular K(+) concentration, similar values were obtained by either technique. The ionspecific microelectrode system was then used to investigate the association between net H(+) efflux and net K(+) influx. Although the computed H(+):K(+) stoichiometry is dependent upon the choice of diffusion coefficients, the values obtained were extremely variable, and net K(+) influx rarely appeared to be charge-balanced by H(+) efflux. In contrast to earlier studies, we found the cortical membrane potential to be highly K(+) sensitive within the micromolar K(+) concentration range. Simultaneous measurements of membrane potential and K(+) influx, as a function of K(+) concentration, revealed similar K(m) values for the depolarization of the potential (K(m) 6-9 micromolar K(+)) and net K(+) influx (K(m) 4-7 micromolar K(+)). These data suggest that K(+) may enter corn roots via a K(+)-H(+) cotransport system rather than a K(+)/H(+) antiporter.

Journal Article↗

PubMed1995

The Termite Gut Microflora as an Oxygen Sink: Microelectrode Determination of Oxygen and pH Gradients in Guts of Lower and Higher Termites.

Clark-type oxygen microelectrodes and glass pH microelectrodes, each with a tip diameter of <=10 (mu)m, were used to obtain high-resolution profiles of oxygen concentrations and pH values in isolated termite guts. Radial oxygen profiles showed that oxygen penetrated into the peripheral hindgut contents up to about 150 to 200 (mu)m below the epithelial surface in both the lower termite Reticulitermes flavipes (Kollar) and the higher termite Nasutitermes lujae (Wasmann). Only the central portions (comprising less than 40% of the total volume) of the microbe-packed, enlarged hindgut compartments ("paunches") were completely anoxic, indicating that some members of the hindgut microbiota constitute a significant oxygen sink. From the slopes of the oxygen gradients, we estimated that the entire paunches (gut tissue plus resident microbiota) of R. flavipes and N. lujae accounted for 21 and 13%, respectively, of the respiratory activity of the intact animals. Axial oxygen profiles also confirmed that in general, only the paunches were anoxic in their centers, whereas midguts and posterior hindgut regions contained significant amounts of oxygen (up to about 50 and 30% air saturation, respectively). A remarkable exception to this was the posterior portion of an anterior segment (the P1 segment) of the hindgut of N. lujae, which was completely anoxic despite its small diameter ((apprx=)250 (mu)m). Axial pH profiles of the guts of Nasutitermes nigriceps (Haldeman) and Microcerotermes parvus (Haviland) revealed that there were extreme shifts as we moved posteriorly from the midgut proper (pH (apprx=)7) to the P1 segment of the hindgut (pH >10) and then to the P3 segment (paunch; pH (apprx=)7). The latter transition occurred at the short enteric valve (P2 segment) and within a distance of less than 500 (mu)m. In contrast, R. flavipes, which lacks a readily distinguishable P1 segment, did not possess a markedly alkaline region, and the pH around the midgut-hindgut junction was circumneutral. The oxic status of the peripheral hindgut lumen and its substantial oxygen consumption, together with previous reports of large numbers of aerobic and facultatively anaerobic bacteria in the hindgut microflora, challenge the notion that termite hindguts are a purely anoxic environment and, together with the steep axial pH gradients in higher termites, refine our concept of this tiny microbial habitat.

Journal Article↗

PubMed1993

Accuracy and precision in vitro of Beetrode microelectrodes used for intraoral pH measurements.

The accuracy and precision of 12 different microelectrodes were assessed through a series of pH recordings of buffer solutions with known pH values. Some electrodes were quite accurate, yielding an average deviation of recordings from true values of about 0.05 pH units, while other electrodes yielded average deviations of up to 0.20 pH units. The precision of the electrodes also varied between electrodes. While all recordings were within 0.15 pH units from the true values for some electrodes, other electrodes showed a marked dispersion with less than 25% of the recordings being within 0.15 pH units from true values. For some electrodes accuracy depended on true pH values, while no such dependency was noted with respect to the precision of the electrodes tested. Both accuracy and precision may be impaired when pH recordings are made on solutions with pH values outside the pH range encompassed by the calibration buffers. Our results demonstrate that the accuracy and precision of these microelectrodes cannot be considered fixed and known parameters. Electrodes should therefore be checked prior to clinical use, and electrodes with low accuracy and precision should be excluded from further use.

Buffers↗

PubMed2006

Fabrication of small complex-shaped optics by plasma chemical vaporization machining with a microelectrode.

We have developed plasma chemical vaporization machining by using a microelectrode for the fabrication of small complex-shaped optical surfaces. In this method, a 0.5 mm diameter pipe microelectrode, from which processing gas is drawn in, generates a small localized plasma that is scanned over a workpiece under numerical computer control to shape a desired surface. A 12 mm x 12 mm nonaxisymmetric mirror with a maximum depth of approximately 3 microm was successfully fabricated with a peak-to-valley shape accuracy of 0.04 microm in an area excluding the edges of the mirror. The average surface roughness was 0.58 nm, which is smooth enough for optical use.

Journal Article↗

PubMed2001

Glucose sensing based on interdigitated array microelectrode.

A micro glucose sensor consisting of an interdigitated array gold microelectrode was developed. The interdigitated array structure, which has 10 microns band width and 10 microns band gap, was fabricated in a small region (2.5 x 5 mm2) on a quartz substrate. Glucose oxidase was chemically fixed onto the electrode surface through self-assembled monolayer of 11-mercaptoundecanoic acid; ferroceneacetic acid was used as electron mediator. Electrochemical properties of the glucose oxidase-immobilized microelectrode were investigated by cyclic voltammogram measurements. Results confirmed that the reductive ferroceneacetic acid generated at counter electrode diffuses through a narrow band gap (10 microns) and can reach the working electrode surface.

Acetic Acid↗

PubMed2000

Predictors of neuropsychological outcome in patients following microelectrode-guided pallidotomy for Parkinson's disease.

OBJECT: The authors studied neuropsychological performance following microelectrode-guided posteroventral pallidotomy in patients with Parkinson's disease (PD) and evaluated correlations with presurgical and surgical factors. METHODS: Neuropsychological changes 3 months (43 patients) and 12 months (27 patients) after microelectrode-guided pallidotomy for PD are reported in a series of 44 consecutive patients with the disease, who improved neurologically, as measured by the Unified Parkinson's Disease Rating Scale (UPDRS) in both the "off' (p<0.001) and best "on" (p<0.001) states. Findings of the vocabulary subtest of the Wechsler Adult Intelligence Scale-Revised (p<0.01), Letter Fluency (p<0.001), Verbal Fluency for semantic categories (p<0.001), and the Wisconsin Card Sorting Test (p<0.01) showed a significant decline in neuropsychological performance in patients 3 months after undergoing left-sided pallidotomy. Impairment in the language domain (semantic fluency) persisted at the 12-month follow-up examination (p<0.01). Visual memory improved after right-sided pallidotomies (p<0.01 after 3 months), with a nonsignificant trend toward persistent improvement 1 year postsurgery (p<0.02 after 12 months). Preoperative semantic fluency was influenced by patient age (p<0.001) and by the width of the third ventricle (p<0.05), as measured by magnetic resonance imaging. A regression model revealed that semantic fluency 3 months postoperatively was significantly affected by the baseline score (p<0.001), side of surgery (p<0.001), handedness (p<0.01), and patient age (p<0.05). However, postoperative lesion volume, lesion location, number of tracks, number of lesions, distance from anatomical landmarks, or UPDRS score did not significantly contribute to neuropsychological outcome. CONCLUSIONS: Neuropsychological changes in a cohort of patients with PD who underwent pallidotomy and experienced excellent clinical benefits and minimum postoperative complications, emphasize the importance of neuropsychological examinations and further investigation of predictive factors.

Aged↗

PubMed2004

Microelectrode recording revealing a somatotopic body map in the subthalamic nucleus in humans with Parkinson disease.

OBJECT: The subthalamic nucleus (STN) is a key structure for motor control through the basal ganglia. The aim of this study was to show that the STN in patients with Parkinson disease (PD) has a somatotopic organization similar to that in nonhuman primates. METHODS: A functional map of the STN was obtained using electrophysiological microrecording during placement of deep brain stimulation (DBS) electrodes in patients with PD. Magnetic resonance imaging was combined with ventriculography and intraoperative x-ray film to assess the position of the electrodes and the STN units, which were activated by limb movements to map the sensorimotor region of the STN. Each activated cell was located relative to the anterior commissure-posterior commissure line. Three-dimensional coordinates of the cells were analyzed statistically to determine whether those cells activated by movements of the arm and leg were segregated spatially. Three hundred seventy-nine microelectrode tracks were created during placement of 71 DBS electrodes in 44 consecutive patients. Somatosensory driving was found in 288 tracks. The authors identified and localized 1213 movement-related cells and recorded responses from 29 orofacial cells, 480 arm-related cells, 558 leg-related cells, and 146 cells responsive to both arm and leg movements. Leg-related cells were localized in medial (p < 0.0001) and ventral (p < 0.0004) positions and tended to be situated anteriorly (p = 0.063) relative to arm-related cells. CONCLUSIONS: Evidence of somatotopic organization in the STN in patients with PD supports the current theory of highly segregated loops integrating cortex-basal ganglia connections. These loops are preserved in chronic degenerative diseases such as PD, but may subserve a distorted body map. This finding also supports the relevance of microelectrode mapping in the optimal placement of DBS electrodes along the subthalamic homunculus.

Brain Mapping↗

PubMed2002

In situ identification of azo dye inhibition effects on nitrifying biofilms using microelectrodes.

In this study, the inhibitory effects of acid orange 7 (AO7), a common azo dye, on nitrification in biofilms were investigated in situ using microelectrodes. Biofilms were obtained from laboratory rotating drum biofilm reactor after the nitrification process reached a pseudo-steady state. Dissolved oxygen, pH, NH4+, NO3-, and redox potential microelectrodes, with tip diameters ranging from 3-15 microm, were used to monitor the spatial distribution and change of microbial activities within nitrifying biofilms. It was found that at lower concentration (1 mg/L), AO7 had only a slight impact on the NH4+-N concentration profiles. The ammonium consumption rate decreased as higher AO7 concentrations (15 mg/L and 25 mg/L) were exposed to the biofilms. A similar trend was observed for the NO3(-)-N microprofiles. The nitrate production rate decreased as the AO7 concentration in the bulk solution increased. The dissolved oxygen and pH microprofiles also showed oxygen and alkalinity utilization, but at lower rates throughout the biofilms when the nitrification process was inhibited. No significant redox potential differences were observed in the biofilms after AO7 was applied.

Azo Compounds↗

PubMed2003

Effect of dissolved oxygen concentration on the biofilm and in situ analysis by fluorescence in situ hybridization (FISH) and microelectrodes.

A better understanding of microbiology and ecology of nitrifying bacteria in inner biofilms is an important part of improving process performance and control. Microelectrodes and fluorescent in situ hybridization (FISH) in biofilm research have been used to investigate the spatial distributions of various microbial activities in biofilms and have led to new experimental findings as well as modifications of the homogeneous assumptions in the biofilm kinetic models. The objective of this study is to try the combination of two methods, both FISH and microelectrode measurements, and to provide reliable and in situ information on nitrifying bacterial activity in biofilms. The characteristics of biofilm developed on tygon slides were different according to the change of dissolved oxygen (DO). When the DO increased from 2 to 10 mg DO/L, the rate of the biofilm thickness increased and its dry density changed from 50-70 to 25-90 mg/cm3. Ammonia oxidizing bacteria were not uniformly distributed in biofilm, and were found at the deeper layer where oxygen is depleted, they were detected primarily in the upper and middle layers of the biofilm.

Bacteria↗

PubMed2004

Effects of hydroxylamine on microbial community structure and function of autotrophic nitrifying biofilms determined by in situ hybridization and the use of microelectrodes.

Effects of hydroxylamine (NH2OH), an intermediate of NH4+ oxidation, on microbial community structure and function of two autotrophic nitrifying biofilms fed with and without NH2OH were analyzed by a 16S rRNA approach and the use of microelectrodes. In the NH2OH-added biofilm, partial oxidation of NH4+ to NO2- was observed, whereas complete oxidation of NH4+ to NO3- was achieved in the control biofilm. In situ hybridization results revealed that no nitrite-oxidizing bacteria (NOB) hybridized with any specific probes were detected in the NH2OH-added biofilm. Thus, the addition of low concentrations of NH2OH (250 microM) completely inhibited the growth of NOB. Phylogenetic analysis of 16S rDNA indicated that the ammonia-oxidizing bacteria (AOB) detected in both biofilms were closely related to Nitrosomonas europaea, and that the clone sequences from both biofilm libraries have more than 99% similarity to each other. However, in situ hybridization results revealed that the addition of NH2OH changed the form of growth pattern of the dominant Nitrosomonas spp. from dense clusters mode to single scattered cells mode. Microelectrode measurements revealed that the average NH4+ consumption rate calculated in the NH2OH-added biofilm was two times higher than that in the control biofilm. This clearly demonstrated that the oxidation of NH4+ was stimulated by NH2OH addition.

Biofilms↗

PubMed1983

Rhythmic afterpotentials in relation to pre-stimulus barbiturate spindle-like activity. Macro- and microelectrode investigations.

It was proposed that the two types of rhythmic afterreactions (RARs) in the somatosensory cortex following forepaw electrical stimulation, i.e. the classical rhythmic afterpotential (RAP) with monophasic waves progressively decreasing in amplitude and the spindle-like afterpotential (SRAP) composed of waxing and waning phase like the spontaneous spindle (SS) were different electrocortical evoked rhythms. The hypothesis that not the RAP, but the SRAP is a SS elicited by stimulation was tested in the present study investigating the relationships between the SS and the two types of RAR by means of macro- and microelectrode recordings. It was shown that the RAP appeared during the SS, while the development of the SRAP was prevented by the SS. The data obtained from the microelectrodes experiments showed that the larger part of the neurons studied discharged in phase both with the waves of the SS and of SRAP (but not of RAP) having an almost one and the same rhythm, supported also the hypothesis for a common pacemaker of the rhythms of the SS and the SRAP.

Animals↗

PubMed1998

Microelectrode measurements of local mass transport rates in heterogeneous biofilms.

Microelectrodes were used to measure oxygen profiles and local mass transfer coefficient profiles in biofilm clusters and interstitial voids. Both profiles were measured at the same location in the biofilm. From the oxygen profile, the effective diffusive boundary layer thickness (DBL) was determined. The local mass transfer coefficient profiles provided information about the nature of mass transport near and within the biofilm. All profiles were measured at three different average flow velocities, 0.62, 1.53, and 2.60 cm sec-1, to determine the influence of flow velocity on mass transport. Convective mass transport was active near the biofilm/liquid interface and in the upper layers of the biofilm, independent of biofilm thickness and flow velocity. The DBL varied strongly between locations for the same flow velocities. Oxygen and local mass transfer coefficient profiles collected through a 70 micrometer thick cluster revealed that a cluster of that thickness did not present any significant mass transport resistance. In a 350 micrometer thick biofilm cluster, however, the local mass transfer coefficient decreased gradually to very low values near the substratum. This was hypothetically attributed to the decreasing effective diffusivity in deeper layers of biofilms. Interstitial voids between clusters did not seem to influence the local mass transfer coefficients significantly for flow velocities of 1.53 and 2.60 cm sec-1. At a flow velocity of 0.62 cm sec-1, interstitial voids visibly decreased the local mass transfer coefficient near the bottom.

Biofilms↗

PubMed2000

Quantitative assay of metronidazole by capillary zone electrophoresis with amperometric detection at a gold microelectrode.

Capillary zone electrophoresis was employed for the determination of metronidazole using end-column amperometric detection with a gold microelectrode at a constant potential of -0.52V vs. saturated calomel electrode. To overcome interference of oxygen in the solution, a deaeration injector and a deaeration protector at the detection cell were used. The optimum conditions of separation and detection are 1.0 x 10(-3) mol/L potassium dihydrogen citrate (KH2C6H5O7) for the buffer solution, 20 kV for the separation voltage, and 5 kV and 10 S for injection voltage and injection time, respectively. The limit of detection is 6.0 x 10(7) mol/L or 0.78 fmole (S/N = 3). The relative standard deviation is 3.9% for the electrophoretic peak current. The method was applied to the determination of metronidazole in human urine.

Electrophoresis, Capillary↗

PubMed1988

Proton channels in snail neurons studied with surface pH glass microelectrodes.

Surface pH was recorded on voltage-clamped snail neurons with Hinke-type glass microelectrodes. During stepwise depolarization from -50 mV to +40 mV the surface pH usually increased at first and then fell rapidly during each 7.5 sec voltage step. The pH changes appeared to be due to proton passage through voltage-sensitive channels. The pH changes were inhibited by cadmium and zinc, and were not as sensitive to external pH as to internal pH. Changes in internal pH lagged behind surface pH. Changes in external calcium had little effect on the surface pH increases seen with small depolarizations. These results confirm earlier conclusions that depolarization opens proton-permeable channels.

Animals↗

PubMed1992

Microelectrode measurements of low frequency electric field effects in cells and tissues.

The average intensities of electric fields induced into tissue can be calculated if the morphology and conductivities of the tissue are known, and such values provide one estimate of dosage for a given field exposure level. However, the microanatomical structures of living tissue, which include gap junctions, tight junctions, highly charged cell coats, and extracellular matrices, as well as complex cell shapes, precludes a detailed characterization of the field and current distribution near the cells which are actually responding to the electric fields. This suggests that a more useful electric field dose metric may be one based on an induced physical effect on the cells. Electric fields have at least three distinct physical effects on cells: the normal plasma membrane potential will be altered; the ionic currents and ion distributions at the extracellular surface will be modified; and mechanical forces will be imposed at the cell surface. Each of these effects can, in principle, be measured through the application of specific microelectrode techniques. Here, the feasibility of using various intracellular and extracellular recording methods to obtain dosimetric values, as well as the contribution these measurements could make to our understanding of electric field interactions with biological tissue, are discussed.

Animals↗

PubMed2006

Monitoring in real time with a microelectrode the release of reactive oxygen and nitrogen species by a single macrophage stimulated by its membrane mechanical depolarization.

Macrophages are key cells of the immune system. During phagocytosis, the macrophage engulfs a foreign bacterium, virus, or particle into a vacuole, the phagosome, wherein oxidants are produced to neutralize and decompose the threatening element. These oxidants derive from in situ production of superoxide and nitric oxide by specific enzymes. However, the chemical nature and sequence of release of these compounds is far from being completely determined. The aim of the present work was to study the fundamental mechanism of oxidant release by macrophages at the level of a single cell, in real time and quantitatively. The tip of a microelectrode was positioned at a micrometric distance from a macrophage in a culture to measure oxidative-burst release by the cell when it was submitted to physical stimulation. The ensuing release of electroactive reactive oxygen and nitrogen species was detected by amperometry and the exact nature of the compounds was characterized through comparison with in vitro electrochemical oxidation of H2O2, ONOO-, NO*, and NO2(-) solutions. These results enabled the calculation of time variations of emission flux for each species and the reconstruction of the original flux of production of primary species, O2*- and NO*, by the macrophage.

Animals↗

PubMed1976

Opossum somatic sensory cortex: a microelectrode mapping study.

Organization of opossum somatic sensory cortex has been investigated utilizing closely spaced microelectrode penetrations (0.25-0.5 mm apart) and delicate mechanical stimulation of body surfaces including the facial vibrissae. Results may be summarized as follows: (1) the general organization of somatic sensory cortex, as originally defined by Lende ('63a) has been confirmed; (2) a double representation of the contralateral mystacial vibrissae and rhinarium, implicit in Lende's original data, was revealed in detail, the two representations being orderly, adjacent, mirror-images of each other; (3) units at a given cortical locus responded to deflection of between one and five mystacial vibrissae, about half responding to movement of a single vibrissa only; (4) about 40% of mystacial vibrissa units showed a directional specificity to the extent that they responded to deflections in only one or two cardinal directions; (5) units located in the medial vibrissa area showed a greater directional specificity than did units located in the lateral vibrissa area; (6) the surface area of rhinarial receptive fields was about ten times the area of first-order rhinarial unit receptive fields (B. Pubols et al., '73); (7) representation of the contralateral forelimb, especially the ventral surface of the forepaw, is extensive, orderly, and precise; (8) representation of the contralateral hindlimb, foot, and tail is minimal, and is confined to the midline convexity; (9) the presence of a small region of bilateral representation, lateral to the regions of contralateral representation, was confirmed. It is suggested that the region of contralateral postcranial representation plus the medial rhinarium and mystacial vibrissa areas are the homologue of SmI in placental mammals, and the region of bilateral representation is homologous to SmII of placental mammals, but that the lateral vibrissa and rhinarium areas are a specialization of somatic sensory cortex unique to the Virginia opossum.

Animals↗

PubMed1986

Flow cytometric estimation of transmembrane potential of macrophages--a comparison with microelectrode measurements.

Potential-dependent accumulation of the lipophilic cationic dye 3,3' dihexyloxacarbocyanine (DiOC6(3)) in macrophages has been investigated. Resulting fluorescence of cells was measured by flow cytometry. Alterations of membrane potential of macrophages were induced by ionophore treatment (valinomycin and gramicidin) in a dose-dependent (10(-5) M-10(-7) M) and time-dependent (0 min-45 min) manner. Resulting changes in relative fluorescence intensity were compared with changes of transmembrane potential measured by intracellular recordings obtained by applying glass microelectrodes. The comparative studies offer the possibility to calibrate the flow cytometric estimate of membrane potential of suspended cells. Equilibration of dye partition between cells and surrounding medium is strictly potential-dependent at dye concentrations between 5 X 10(-8) M and 10(-7) M and within an incubation interval from 10 min up to 30 min after addition of dye. Conclusions are drawn concerning the field of application of the optical method. Dynamics of electrical processes following ionophore treatment are discussed in terms of molecular mechanisms of altered ionic transport.

Animals↗