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Glass microelectrodes filled with a saturated solution of methyl blue or fast green in 1.0M potassium acetate can be used to mark penetrated ne. urons from which intracellular recordings have been made. Many cells can be marked in one experiment and can easily be located in subsequent histologic section.
A novel oxygen microelectrode with a tip diameter of 2 to 20 mum was constructed which could function satisfactorily under a variety of environmental conditions and in a variety of media, including human blood serum, citric acid at pH 2.5, moist air, and paraffin oil. Measurement of oxygen by this electrode does not require stirring of the medium. Electrodes could be made so that the 90% response time necessary to detect changes in oxygen concentration was less than 0.2 s, and response was linear with oxygen concentration. To demonstrate the performance of the electrode, oxygen and photosynthesis profiles of an acid microbial mat (pH 2.8) dominated by the eucaryotic alga Cyanidium caldarium were made. Photosynthetic rates as high as 95 mmol of O(2) dm h were measured within the most active 0.1-mm layer, which was ca. 0.2 mm below the surface of the microbial mat. The total photosynthetic activity was 47 mmol of O(2) m h. Vertical profiles of photosynthesis at different light intensities showed that the microalgae within the mat were not photoinhibited at bright sunlight (2,090 muEinsteins m s).
Microelectrodes were used to measure oxygen, pH, and oxygenic photosynthetic activity in a hot spring microbial mat (Octopus Spring, Yellowstone National Park), where the cyanobacterium Synechococcus lividus and the filamentous bacterium Chloroflexus aurantiacus are the only known phototrophs. The data showed very high biological activities in the topmost layers of the microbial mat, resulting in extreme values for oxygen and pH. At a 1-mm depth at a 55 degrees C site, oxygen and pH reached 900 muM and 9.4, respectively, just after solar noon, whereas anoxic conditions with a pH of 7.2 were measured before sunrise. Although diurnal changes between these extremes occurred over hours during a diurnal cycle, microbial activity was great enough to give the same response in 1 to 2 min after artificial shading. Oxygenic photosynthesis was confined to a 0.5- to 1.1-mm layer at sites with temperatures at or above about 50 degrees C, with maximum activities in the 55 to 60 degrees C region. The data suggest that S. lividus is the dominant primary producer of the mat.
Beggiatoa spp. grow optimally in media containing opposed gradients of oxygen and soluble sulfide, although some strains also require an organic substrate. By using microelectrodes, we characterized oxygen and sulfide gradients during their initial development in uninoculated media and in cultures of marine and freshwater strains. In gradient media, Beggiatoa strains always grew some distance below the air/agar interface as a dense "plate" of constantly gliding filaments with sharply demarcated upper and lower boundaries. Within established plates, the maximum oxygen partial pressure was 0.6 to 6.0% of air saturation and not significantly lower if filaments were fixing nitrogen. Oxygen penetrated only 100 to 300 mum into the plate, and the anoxic fraction increased from less than 10% to approximately 90% during later stages of growth. For lithoautotrophically grown marine strains, the linearity of the oxygen profile above the plate plus its drop to zero therein indicated that oxygen uptake for the entire tube occurred only within the Beggiatoa plate. Consequently, oxygen consumption could be predicted solely from the distance between the air/agar interface and the top of a plate, given the diffusion coefficient for oxygen. By contrast, for freshwater strains grown heterotrophically (with sulfide also in the medium), oxygen profiles were frequently nonlinear because of nonbiological reaction with sulfide which had diffused past the aggregated filaments. For all strains tested, microoxic aggregation also occurred in the absence of sulfide, apparently reflecting a step-up phobic response to oxygen.
NO(3) concentration profiles were measured in the sediments of a meso-eutrophic lake with a newly developed microelectrode. The depth of penetration of NO(3) varied from only 1.3 mm in organic-rich profundal silty sediments to 5 mm in organic-poor littoral sandy sediments. The thickness of the zone of denitrification in the organic-rich sediments was <500 mum. Oxygen profiles measured simultaneously revealed that the zone of denitrification was directly adjacent to the aerobic zone. The results demonstrate high denitrification rates (0.26 to 1.31 mmol m day) at in situ nitrate concentrations in the overlying water (0.030 mmol liter) and limitation of denitrification by nitrate availability.
Cutaneous afferent activity was recorded in fascicles of the median and radial nerves of normal subjects using percutaneous microelectrodes. Multi-unit fascicular responses were dominated by activity in large myelinated fibres. Easily tolerated electrical stimuli evoked the full spectrum of fast and slow myelinated fibre activity but more intense levels were required to activate unmyelinated fibres. Correlation of the evoked potentials and the sensations reported by the subject suggested that fast myelinated fibres mediate tactile sensations. Pricking pain appeared with the activation of slow myelinated fibres. The only sensations induced by electrical stimuli were tactile or painful.
Eight of the first 15 patients with advanced Parkinson's disease who underwent microelectrode guided posteroventral pallidotomy developed transient abnormal involuntary movements during thermolesion, four of whom also did so during high frequency macrostimulation. Abnormal involuntary movements found before thermolesion were choreic, ballistic, or choreoathetoid in nature, usually persisted less than 60 minutes, and were contralateral to the site of thermolesion in six and bilateral in two of them. The appearance of abnormal involuntary movements during macrostimulation or thermolesion of the internal globus pallidus correlated with better surgical outcome as measured by UPDRS motor items and CAPIT timed test, so that they seem to be of prognostic value.
Ion-selective microelectrodes (ISMs) were used to measure the turnover of intracellular K+ (Ki+) in rods in the isolated retina of the toad, Bufo marinus. The light-evoked hyperpolarization of rods decreases their passive K+ efflux, which in combination with active K+ uptake, decreases extracellular K+ concentration, Ko+.Rb+ substitutes for K+ in these processes. The turnover of Ki+ was measured as Rb+ and K+ were exchanged, using ISMs that were approximately five times more sensitive to Rb+ than to K+. When Ko+ was replaced by Rbo+, the light-evoked decrease in K+ efflux produced only a small change in ISM voltage, delta VISM, owing to the background of Rbo+. As Rbi+ replaced Ki+, the efflux shifted from K+ to Rb+ and delta VISM grew in amplitude. After loading the rods with Rbi+, Rbo+ was replaced by Ko+. The light-evoked decrease in Rb+ efflux lead transiently to a large delta VISM, since the change in Rbo+ was superimposed upon a background of Ko+. As Ki+ replaced Rbi+, the amplitude of delta VISM declined. When measured using this technique, the turnover of Ki+ was 95% complete in approximately 15 min. In low Ca2+ solutions, transmembrane fluxes of K+ (Rb+) increased and turnover of Ki+ occurred more rapidly. During background illumination, transmembrane fluxes of K+ (Rb+) decreased and turnover of Ki+ was slowed. These experiments have provided independent corroboration of earlier observations concerning rod K+ fluxes. This ISM-based technique also may be useful in measuring K+ turnover in other cell types.
Using pH-sensitive microelectrodes, we evaluated pH dynamics of extracellular fluid (ECF) within the ventrolateral medulla (VLM) beneath the central chemoceptive areas in anesthetized, spontaneously breathing cats. Static ECF pH was acid in the superficial layers (less than 1 mm), compared with the overlying cerebrospinal fluid pH that became alkaline gradually during the experiments. In the deeper VLM areas (1-3 mm), no systematic gradients of ECF pH were observed. We found various, isolated regions where intravertebral artery injections of CO2-saturated saline evoked acidic shift of ECF pH in the time course analogous to ventilatory augmentation. Those responsive regions were found to be scattered not only in the superficial layers but also in the deeper VLM areas, although many nonresponsive regions were also intermingled among them. Occlusions of the principal vessels supplying the tested VLM regions diminished but failed to abolish the ECF pH responses to the CO2 loadings, suggesting a collateral blood flow by fine pial vessels. The present study suggests a possibility that the pH-dependent central chemoreceptors, if any, would be scattered in the deeper VLM areas as well as the superficial layers.
Periods of tachycardia were induced in isolated segments (15 X 15 mm) of rabbit left atrium by local application of a properly timed premature stimulus. We used a special device for multiple synchronous microelectrode recordings of responses of more than 100 fibers during the initiation of tachycardia. We clearly demonstrated circus movement of the impulse through a small area of atrial muscle as the underlying mechanism. The premature impulse was conducted antegrade in only one direction, whereas in the other directions antegrade conduction failed. The local responses of the fibers in the blocked area served as a temporary obstacle for return of the premature impulse. When these fibers recovered their excitability before extinction of the premature impulse, they were reentered in a retrograde direction, and the impulse traveled in a circular route. During the propagation of a premature beat, local block, which set the stage for circus movement, was caused by nonuniform recovery of excitability of the atrium. We related the spread of activation of a premature impulse to the naturally occurring spatial dispersion in refractory periods and found that local conduction block invariably was associated with an area of delayed restoration of excitability. Artificial induction of differences in refractory periods by regional application of carbamylcholine made it clear that a disparity in refractory periods of only 11-6 msec between adjacent areas may be sufficient to cause local conduction block of a properly timed premature impulse.
Intracellular microelectrode recordings were made from smooth muscle cells in cross-sectional preparations of equine ileum, superfused in vitro. Membrane potential oscillations and spike potentials were recorded in all preparations, but recordings were made more readily from cells in the longitudinal muscle layer than from cells in the circular layer. The mean (se) resting membrane potential (RMP) of smooth muscle cells in the longitudinal muscle layer was -51.9 (1.2) mV, and the membrane potential oscillations in this layer had a mean amplitude of 4.8 (0.4) mV, a frequency of 9.0 (0.1) cycles per minute and a duration of 5.8 (0.2) seconds. The membrane potential oscillations were preserved in the presence of tetrodotoxin. A waxing and waning pattern of membrane potential oscillation activity was observed. Nifedipine abolished the spiking contractile activity of the smooth muscle, did not abolish the membrane potential oscillations but did alter their temporal characteristics.
A simple method was suggested which enabled to estimate the intercapillary PO2 under different conditions. The intercapillary distribution of the cortex tissue oxygen pressure in normal conditions and in hypoxia was studied in unanesthetized and immobilized cats with the aid of microelectrodes. The absolute values obtained varied from 1 to 95 mm Hg. Initially high PO2 underwent maximum changes in hypoxia whereas lower PO2 values did not change significantly.
The phylogenetic diversity of a nitrifying bacterial community of two types of nitrifying biofilms, a domestic wastewater biofilm and an autotrophic nitrifying biofilm grown on rotating disk reactors (RDR), was characterized by 16S ribosomal DNA (rDNA)-cloning analysis. Thereafter, successional development of nitrifying the bacterial community within both biofilms was visualized in situ by fluorescent in situ hybridization (FISH) wih a set of fluorescently labeled 16S rRNA-targeted DNA probes. In situ hybridization revealed that Nitrosomonas ureae was the numerically dominant species of the ammonia-oxidizing population in the domestic wastewater biofilm and that a population shift from N. urea to N. europaea and N. eutropha occurred when the culture medium was switched to the synthetic media from the domestic wastewater. After reaching the steady-state condition, microprofiles of NH4+, NO2-, NO3-, and O2 in the biofilms were measured by use of microsensors, and the spatial distributions of in situ nitrifying activities were determined. The relationship between the spatial organization of nitrifying bacterial populations and the in situ activity of these populations within the biofilms was discussed. Microelectrode measurements revealed that the active ammonia-oxidizing zone was vertically separated from the active nitrite-oxidizing zone. This vertical separation became more evident with increase of the substrate C/N ratio, leading to deterioration of nitrification efficiency. The combined use of these techniques made it possible to relate in situ nitrifying activity directly to the occurrence of nitrifying bacterial populations.
A versatile microelectrode array sensor for water quality monitoring has been developed. The array fabrication, based on batch microelectronic processes, results in a highly stable passivation of the silicon chip surface and provides the possibility to use a backside contact. Packaging was optimized for on-line water operation at high pressures. Examples of applications include chlorine monitoring in drinking water, ozone monitoring in deionized water, dissolved oxygen in activated sludge and preliminary measurements of trace arsenic.
Tension, dT/dt, and extracellular free [Ca] [( Ca]0) were continuously measured in isolated rabbit right papillary muscles at 28 degrees C. Double-barreled Ca-selective microelectrodes incorporating Simon's neutral Ca exchange resin (ETH-1001) were used to monitor [Ca]0. Decreases of [Ca]0 were seen during the course of single twitches, before the development of significant tension. This depletion of Ca0 probably represents Ca influx into the cells from the extracellular space. The magnitude of this Ca0 depletion is decreased by Co, verapamil, long rest intervals, and reduction of [Ca]0. The magnitude is increased by catecholamines, reduction of [Na]0, caffeine, continued pacing, and elevation of [Ca]0. After 3-min rest, stimulation (0.5-1 Hz) produces a biphasic tension response (larger first beat, small second beat, and monotonic rise to control). Caffeine (5 mM) changes the pattern after rest to a monotonic increase. Ca influx increases monotonically in both cases. Addition of 20 mM Co during the rest reduces tension of all beats by similar amounts in the presence of caffeine. In the absence of caffeine, Co has a much weaker effect on the first beat than on subsequent beats. The results suggest that caffeine inhibits an intracellular component of activator Ca that is more important after a rest interval, but that Ca influx becomes increasingly more important during continued pacing under the conditions used here.
Spikes and postsynaptic potentials (PSPs) were recorded in the sensorimotor cortex of awake rabbits. (1) A cellular analog of cortico-cortical conditioned reflex (CR) was studied. Direct cortical stimulation of a remote point (conditional stimulus - CS) was paired with the stimulation near the microelectrode insertion (unconditional stimulus - UCS). The most neurons showed response facilitation when short intertrial intervals and strong UCS were used. These changes were short-lasting (up to 30 s) and were explained by posttetanic potentiation. Of 21 units tested with long (7 to 120 s) intertrial intervals, 9 showed significant response modification of longer duration. (2) The analog of cortico-cortical CR was modified using stimulation of homolateral cortex as the CS. The UCS was complimented by lateral hypothalamic reinforcement (LHR) in instrumental or classical paradigm. Of 37 units tested, 5 showed an increase in the probability of short-latency ( less than 25 ms) spike. (3) Extracellular activity of 17 neurons was followed through the entire cycle of elaboration and extinction of a "local conditioned startle response" established by pairing click CS with cortical UCS and LHR. Responses of 7 neurons showed significant modifications. The latency distributions of averaged PSPs were compared for naive (N), conditioned (C) and extinguished (E) states. Latencies in N and E groups were similar but were different from those in C group. Of 41 C group neurons, 26 responded at a latency of less than 17 ms; and 9 responded after less than or equal to 7 ms. A pathway for this simple CR may pass through the cortex. Thus PSP changes were observed in all models. Augmentation of short-latency EPSPs support the idea that an increase in efficacy of excitatory synapses underlies the conditioning. Interneuronal excitatory connections within the sensorimotor cortex are, presumably changed during elaboration of simplest motor CRs. These connections can be analyzed with intracellular recordings triggered by spikes of a neighbouring neuron.
The Electronic Clinical Brain Atlas is a CD-ROM containing several classic brain atlases and a real-time yet simple registration function that deforms the atlases to match them with specific patient studies. This article presents the use of this registration function for functional neurosurgery planning. We first propose the CD-ROM assisted planning procedure, then illustrate it with two cases: a pallidal stimulation and a thalamic stimulation. The Schaltenbrand-Wahren atlas is registered and scaled to conform with an actual patient's data by means of two-dimensional (2-D) local deformations performed in multiple orientations. First a rectangular region of interest (ROI), which is set between any clearly visible landmarks chosen by the neurosurgeon, is measured on the film or scanner console. The corresponding atlas plate with the target is then deformed in real time for the same landmarks, such that the dimensions of this ROI are the same on the film and on the deformed atlas plate. Next the target is set on the deformed (individualized) atlas plate and its coordinates are read. The individualized atlas plate can also be printed on transparent foil and superimposed on the film or, alternatively, this superimposition can be done electronically. The planning steps can be repeated for all available orientations. The proposed atlas-assisted planning procedure extends the traditional use of printed stereotactic atlases by individualizing them to specific patients. The preliminary results show that this procedure may improve the definition of the target and may have several advantages over other approaches, such as indirect measurements based on the AC-PC line or 1-D (intercommissural distance based) scaling. It provides the neurosurgeon with a convenient and immediate means of accessing ancillary data that is usually only available in the printed form.
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