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PubMed1978

Effects of amphotericin b on the electrical properties of Necturus gallbladder: intracellular microelectrode studies.

Intracellular microelectrode techniques were employed to study the mechanism by which amphotericin B induces a transient mucosa-negative transepithelial potential (deltaVms) in the gallbladder of Necturus. When the tissue was incubated in standard Na-Ringer's solution, the antibiotic reduced the apical membrane potential by about 40 mV, and the basolateral membrane potential by about 35 mV, whereas the transepithelial potential increased by about 5 mV. The electrical resistance of the apical membrane fell by 83%, and that of the basolateral membrane by 40%; the paracellular resistance remained unchanged. Circuit analysis indicated that the equivalent electromotive forces of the apical and basolateral membranes fell by 35 and 11 mV, respectively. Changes in potentials and resistances produced by ionic substitutions in the mucosal bathing medium showed that amphotericin B produces a nonselective increase in apical membrane small monovalent cation conductance (K, Na, Li). In the presence of Na-Ringer's on the mucosal side, this resulted in a reduction of the K permselectivity of the membrane, and thus in a fall of its equivalent emf. During short term exposure to amphotericin B, PNa/PCl across the paracellular pathway did not change significantly, whereas PK/PNa doubled. These results indicate that deltaVms is due to an increase of gNa across the luminal membranes of the epithelial cells (Cremaschi et al., 1977. J. Membrane Biol. 34:55); the data do not support the alternative hypothesis (Rose & Nahrwold, 1976. J. Membrane Biol 29:1) that deltaVms results from a reduction in shunt PNa/PCl acting in combination with a rheogenic basolateral Na pump.

Amphotericin B↗

PubMedDate not supplied

Intracellular sodium activity and Bretschneider's cardioplegia: continuous measurement by ion-selective microelectrodes at initial equilibration.

Intracellular sodium activity (aiNa), intracellular pH (pHi) and membrane potential were directly and continuously measured in sheep cardiac Purkinje fibers using neutral carrier liquid membrane ion-selective microelectrodes. Changing the superfusing medium from normal Tyrode's solution to the cardioplegic solution "HTK" according to Bretschneider (6) a depolarization from -73.7 +/- 7.2 mV to -55.0 +/- 9.5 mV (n = 25), a decrease of aiNa from 9.1 +/- 1.9 mM to 4.0 +/- 1.4 mM (n = 25) and an intracellular acidification from pHi 7.18 +/- 0.06 to pHi 7.01 +/- 0.06 (n = 11, mean +/- S.D.) occurred at 35 degrees C. The decrease of intracellular sodium activity was not effected by replacement of K, Mg, or histidine by mannitol in the cardioplegic solution. Addition of 4 mM Ca somewhat enhanced aiNa decline. Inhibition of the sodium pump with the cardiac steroid dihydroouabain (10(-4) M) lowered the effect of "HTK" on intracellular sodium by approximately 35% (n = 5). Sodium decline was also sensitive to equilibration temperature, giving a Q10 of 1.54 for the initial decrease velocity (temperature range 20 to 35 degrees C), which is less than that found by other investigators for pure sodium pump activity. It is suggested that although the electrochemical sodium gradient remains inward throughout, sodium may leave myocardial cells on induction of Bretschneider's cardioplegia because of a reduction of inward fluxes by simultaneously increasing sodium pump activity, thus increasing Na efflux. Na/Ca exchange is assumed to be of minor importance and the Na/H exchange may be involved. With respect to the clinical application of the low Na and nominally Ca-free cardioplegic solution "HTK" lowering of intracellular sodium activity is interpreted as a factor minimizing the risk of a "calcium paradox" on reperfusion with Ca at serum levels, as well as a possible mechanism preventing early development of cellular edema.

Animals↗

PubMed1983

Transmembrane potential of J774.2 mouse macrophage cells measured by microelectrode and ion distribution methods.

The transmembrane potential (Em) of J774.2 macrophage cells measured by microelectrodes was -24.1 +/- 0.7 mV (mean +/- SEM). Em measured by lipophilic ion distribution was -35 +/- 2 mV or -40 +/- 2 mV, using a cation or anion, respectively. By any method, colchicine reduced Em by approximately 3 mV.

Animals↗

PubMed1987

The effects of chloride ions on electrodiffusion in the membrane of a leaky epithelium. Studies of intact tissue by microelectrodes.

The electrodiffusive permeability for Cl-, its dependence on low extracellular Cl--concentrations and the interaction between the movements of Cl- and K+ were investigated in the ventricular membrane of epithelial cells from the choroid plexus of Necturus maculosus. Cells were probed with ion-selective microelectrodes sensitive to Cl-, K+ and H+. The initial effects of abrupt changes in the Cl--concentration (Cl-v) and/or the K+-concentration (K+v) of the ventricular solution were investigated. The effect of changing the membrane potential by changing K+v was twofold: It caused an electrodiffusive flux of Cl- via a permeability of 1.3 X 10(-6) cm s-1. This permeability together with the K+-permeability of the ventricular membrane (24 X 10(-6) cm s-1) determined the membrane potential in the given steady state within a few mV. The other effect of the depolarization was an increase in the intracellular concentration of HCO-3 which in turn caused an influx of Cl- via electroneutral Cl-/HCO-3 exchange. The Cl--permeability was reduced by more than 60% and the neutral exchange by more than 90% by furosemide. The effect of decreases in Clv was a tenfold increase of the electrodiffusive Cl--permeability of the ventricular membrane to 12.2 X 10(-6) cm s-1 and also a tenfold increase in the permeability to K+. This activation was reduced by two thirds by furosemide, and by depolarizations of the cell by high K+v. In the given steady state the HCO-3/Cl- exchanger at the ventricular membrane transports at a rate of 300 pmol cm-2 s-1 and moves Cl- into the cell and HCO-3 into the ventricular solution. Thus the epithelium alkalinizes the cerebrospinal fluid at a rate which is about three times faster than the net transport rate of Na+.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗

PubMed2000

Silicon-based microelectrodes for neurophysiology, micromachined from silicon-on-insulator wafers.

A process is described for the fabrication of silicon-based microelectrodes for neurophysiology using bonded and etched-back silicon-on-insulator (BESOI) wafers. The probe shapes are defined without high levels of boron doping in the silicon; this is considered as a step towards producing probes with active electronics integrated directly beneath the electrodes. Gold electrodes, of 4 microns by 4 microns to 50 microns by 50 microns are fabricated on shanks (cantilever beams) 6 microns thick and which taper to an area approximately 100 microns wide and 200 microns long, which are inserted into the tissue under investigation. The passive probes fabricated have been successfully employed to make acute recordings from locust peripheral nerve.

Animals↗

PubMed1998

Preliminary study on the suitability of a pharmacological bio-assay based on cardiac myocytes cultured over microfabricated microelectrode arrays.

There are a range of techniques that can be used to assay bioactive compound. One potentially promising technique is a system consisting of microfabricated extracellular recording devices over which electrogenic cells can be grown. To date, research in this area has concentrated on the use of neurons as an electrogenic cell type. However, these cells have limitations. Only small extracellular potentials have been recorded from mammalian neurons cultured over microfabricated electrode arrays. Although such potentials may be of use in assays examining the effects of bio-active compound analogues on firing frequency, they are of little use for more detailed pharmacological studies involving analyses of signal shape. What is required is a system from which much larger extracellular potentials can be recorded. This preliminary study reports on a system based on cardiac myocytes cultured over microfabricated metal microelectrode arrays, from which potentials with a mean amplitude of 16.9 microV can be reliably recorded, which can be reversibly blocked with mumoll-1 concentrations of the sodium ion channel blocker lidocaine. Less common potentials with amplitudes of up to 3.5 mV were also recorded. It is demonstrated that cardiac myocytes cultured over microfabricated micro-electrode arrays can be used in assays of cardioactive compound analogues.

Animals↗

PubMed2000

In vivo measurements of the internal pH of Hediste (Nereis) diversicolor (Annelida, Polychaeta) exposed to ambient sulphidic conditions using pH microelectrodes.

The effect of different ambient sulphide concentrations on the internal pH regime of Hediste (Nereis) diversicolor was studied under in vivo conditions using liquid membrane pH microelectrodes, a method which is new to marine sciences. As a case study, the hypothesis was tested whether organisms exposed to ambient sulphidic conditions are able to lower their internal pH which, in effect, would reduce sulphide influx into the animals and thus could represent an effective detoxification mechanism. It was shown that a significant lowering of the internal pH occurred within only 20 min after adding sulphide. This pH lowering appeared to be dependent on the external sulphide concentration of the ambient medium and showed a saturation beyond a threshold level of about 130 microM. It is discussed whether this sulphide-induced pH drop is an active regulatory mechanism and acts as an effective protection mechanism against sulphide during short-term exposures.

Animals↗

PubMed2003

Biological application of microelectrode arrays in drug discovery and basic research.

Electrical activity of electrogenic cells in neuronal and cardiac tissue can be recorded by means of microelectrode arrays (MEAs) that offer the unique possibility for non-invasive extracellular recording from as many as 60 sites simultaneously. Since its introduction 30 years ago, the technology and the related culture methods for electrophysiological cell and tissue assays have been continually improved and have found their way into many academic and industrial laboratories. Currently, this technology is attracting increased interest owing to the industrial need to screen selected compounds against ion channel targets in their native environment at organic, cellular, and sub-cellular level. As the MEA technology can be applied to any electrogenic tissue (i.e., central and peripheral neurons, heart cells, and muscle cells), the MEA biosensor is an ideal in vitro system to monitor both acute and chronic effects of drugs and toxins and to perform functional studies under physiological or induced pathophysiological conditions that mimic in vivo damages. By recording the electrical response of various locations on a tissue, a spatial map of drug effects at different sites can be generated, providing important clues about a drug's specificity. In this survey, examples of MEA biosensor applications are described that have been developed for drug screening and discovery and safety pharmacology in the field of cardiac and neural research. Additionally, biophysical basics of recording and concepts for analysis of extracellular electrical signals are presented.

Action Potentials↗

PubMed2005

The detection of formaldehyde in textiles using interdigitated microelectrode array diffusion layer titration with electrogenerated hypobromite.

An interdigitated microelectrode array (IDA) was applied to the determination of formaldehyde released from textiles produced in industry. The proposed method is based on formaldehyde reaction with hypobromite which is formed in weakly basic media by control current electrooxidation of bromide on the generator segment of the IDA array. The unreacted hypobromite diffuses through the gap between individually polarisable IDA segments and it is amperometrically detected on the collector segment of the IDA. The efficiency of this nonconvective transfer process in the absence of formaldehyde was substantially higher (78%) in comparison with that when using the rotating ring disc electrode. The influence of the added formaldehyde on the transfer process can be utilised to develop a simple and sensitive analytical procedure for formaldehyde detection with a detection limit of 4 x 10(-6) mol dm(-3).

Bromine Compounds↗

PubMed2005

Electroanalytical method for determination of the pesticide dichlorvos using gold-disk microelectrodes.

This paper reports the use of laboratory-prepared gold microelectrodes and square-wave voltammetry for analytical determination of low concentrations of the pesticide dichlorvos in pure and natural water samples. After optimization of the experimental and voltammetric conditions, the best voltammetric responses-current intensity and voltammetric profile-were obtained in 0.1 mol L(-1) NaClO4 with f=100 s(-1), a=50 mV, and DeltaE(s)=2 mV. The observed detection and quantification limits in pure water were 7.8 and 26.0 microg L(-1), respectively. The reproducibility and repeatability of the method were also determined; the results were 1.4% (n=5) and 1.2% (n=10), respectively. Possible interfering effects were evaluated in natural water samples collected at different points with different levels of contamination from agricultural, domestic, or industrial waste from an urban stream. Results showed that the detection and quantification limits increased as a function of the quantity of organic matter present in the samples. Nonetheless, the values observed for these method characteristics were below the maximum value allowed by the Brazilian code for organophosphorus pesticides in water samples. Recovery curves constructed using the standard addition method were shown to be satisfactory compared with those obtained from high-performance liquid chromatography, confirming the suitability of the method for analysis of natural water samples. Finally, when the method was used to determine dichlorvos in spiked cows' milk samples, satisfactory recovery and relative standard deviations were obtained.

Animals↗

PubMed2005

Characteristics of ischaemic human myocardium: a transoesophageal echocardiographic and voltammetric microelectrode study.

Delayed local myocardial power development (primary asynchrony) has been suggested as a marker of ischaemic ventricular dysfunction in humans. However, to prove this, microcirculatory perfusion, microcirculatory oxygenation, and intrinsic mechanical function of the same asynchronous myocardial segment should be studied simultaneously before and after revascularisation. We performed a prospective intraoperative study of 15 patients (age 67 [SD 5] years) at baseline and 30 min after left anterior descending artery grafting. Local tissue perfusion and oxygenation of the anterior left ventricular wall were quantified with a voltammetric microelectrode technique. Transesophageal M-mode echocardiograms and simultaneous high-fidelity left ventricular pressure were measured. Eight patients showed primary asynchrony and 7 did not. Patients with primary asynchrony had local mechanical depression with lower resting values of myocardial work and peak power which increased with surgery. In this group, resting perfusion consistently increased with surgery (32.1 [13] to 54 [31] ml min(-1) 100 g(-1), P < 0.05). In the remaining patients, local work and power were normal, and resting perfusion was consistently higher (90 [9] Ml min(-1) 100 g(-1), P < 0.05 vs primary asynchrony), and fell with surgery. Local tissue oxygen tension was similar in both groups (38 vs 44 mmHg) and did not change with surgery. In patients with chronic coronary artery disease, microcirculatory perfusion, but not pO2, is reduced in regions showing primary asynchrony and impaired mechanical function. Abnormalities in both mechanical function and perfusion normalise within 30 min of revascularisation. These data provide further evidence that primary asynchrony is not only a marker of chronic ischemic ventricular dysfunction, but is associated with a modified contraction pattern in which normal oxygen tension coexists with reduced perfusion.

Aged↗

PubMed2006

Fabrication and testing of microelectrodes for small-field cortical surface recordings.

A microfabrication approach to produce a microelectrode array that is suitable for use with human patients has been developed. The device is comprised of materials that are consistent with those of clinically used macroelectrodes (platinum electrode contacts suspended within a biomedical grade polydimethylsiloxane, PDMS). Photolithography, metal deposition, wire bonding, and PDMS encapsulation were used to fabricate the device. Cytotoxicity testing with both mammalian and human cortical cells suggests that the device is suitable for use with human patients and implementation of the device in animal studies revealed that reliable evoked potentials could be acquired with the designed spatial resolution.

Action Potentials↗

PubMed2006

Application of a PDMS microstencil as a replaceable insulator toward a single-use planar microelectrode array.

Here we present a novel idea for a replaceable insulator, and thus advance toward the goal of a single-use planar microelectrode array (MEA) for the study of electrogenic tissues. The concept of a replaceable insulator is motivated by insulator degradation after repeated usage of an MEA. Instead of fabricating a more durable insulator for repeated MEA usage, we propose replacing the insulator and effectively producing a fresh MEA for each experiment. We chose a polydimethylsiloxane (PDMS) microstencil as a candidate for the replaceable insulator as it is biocompatible, shows reversible adhesion to surfaces, and can be easily and controllably fabricated. As a proof-of-concept, we demonstrate two applications using microstencils: the rejuvenation of an old MEA and the fabrication of a single-use MEA. These MEAs were tested with dissociated neural cell cultures and neural recordings were performed at 14 days in vitro. Inexpensive and quick supply of insulators with micrometer-sized holes provides a way of constructing an MEA that can be treated as a disposable component in high throughput cell-based biosensor applications.

Animals↗

PubMed2006

An automatic algorithm for stationary segmentation of extracellular microelectrode recordings.

Extracellular microelectrode recordings (MER) often contain artifact from a variety of sources that confound traditional signal-processing techniques that require stationary signal segments. We designed an algorithm to locate the longest stationary segment of MER signals. In this paper we provide a description of the segmentation algorithm and its performance assessment. Simulation results demonstrate that the automatic segmentation algorithm we proposed is capable of accurately identifying the boundaries of the longest stationary segments in MER signals. In our simulation study the segmentation algorithm correctly identified the boundaries of the longest MER stationary segments in 99.5% of the cases.

Algorithms↗

PubMed1986

pH microelectrode: modified Thomas recessed-tip configuration.

Through the use of a glass-membrane pH electrode and a water-tight seal a modified Thomas pH microelectrode has been developed. The modified Thomas electrode has a relatively low electrical resistance (10(11) omega), a small sensing chamber (10 microns3), and a rapid response time (10 s) and can be manufactured in both single- and double-barreled configurations. The modified Thomas electrode is designed to measure the intracellular pH of small cells such as those found in the mammalian kidney tubule.

Animals↗

PubMed1986

Sodium-sensitive glass microelectrode: modified Thomas recessed-tip configuration.

Using a glass-membrane, sodium-sensitive microelectrode, a modified Thomas sodium-sensitive electrode has been developed. The modified Thomas electrode possesses a high sensitivity (57.9 mV/log aNa), a high selectivity (KpotNa,K less than 0.005), a relatively low electrical resistance (7.65 X 10(11) ohms), a small sensing chamber (10 microns3), and can be made in the double-barreled configuration. The modified Thomas electrode is designed to directly measure the intracellular sodium concentration of epithelial cells.

Glass↗

PubMed1987

Microelectrode of the Thomas type using a liquid membrane electrode.

By replacing the glass-based pH electrode (L. R. Pucacco, S. K. Corona, H. R. Jacobson, and N. W. Carter (1986) Anal. Biochem. 153, 251-261) with a liquid membrane-based pH electrode, a relatively easy-to-manufacture modified Thomas electrode has been developed. The liquid membrane-based modified Thomas electrode can be manufactured without the special equipment (forge) and materials (glass) required to make the glass membrane pH microelectrode (L. R. Pucacco and N. W. Carter (1976) Anal. Biochem. 73, 501-512). The sensitivity (57.4 +/- 0.22 mV/pH unit), response time (20.0 +/- 2.67 s), and electrical resistance (3.48 +/- 0.67 X 10(11) ohm) of this electrode are similar to those of the glass-based version.

Electronics, Medical↗

PubMed1988

An ammonia-sensing air gap microelectrode.

An ammonia-sensing air gap microelectrode has been designed on the basis of a neutral carrier pH-sensing inner electrode. This electrode has a tip diameter of 2 to 5 microns, has a simple design, is easy to fabricate, and has a long shelf life. Its response to ammonium is linear in the range 3 x 10(-5) to 10(-2) M and its response time (95%) is 10 to 15 s. The electrode was converted to a microsensor for urea by immobilization of urease within its tip. The linear response to urea ranged from 3 x 10(-4) to 10(-2) M and the response time was 15 to 20 s.

Ammonia↗