Dielectrophoretic trapping of dissociated fetal cortical rat neurons

Recording and stimulating neuronal activity at multiple sites can be realized with planar microelectrode arrays. Efficient use of such arrays requires each site to be covered by at least one neuron. By application of dielectrophoresis (DEP), neurons can be trapped onto these sites. This study investigates negative dielectrophoretic trapping of fetal cortical rat neurons. A planar quadrupole microelectrode structure was used for the creation of a nonuniform electric field. The field was varied in amplitude (1, 3, and 5 V) and frequency (10 kHz-50 MHz). Experimental results were compared with a theoretical model to investigate the yield (the number of neurons trapped in the center of the electrode structure) with respect to time, amplitude and frequency of the field. The yield was a function of time1/3 according to theory. However, unlike the model predicted, an amplitude-dependent frequency behavior was present and unexpected peaks occurred in the DEP-spectra above 1 MHz. Gain/phase measurements showed a rather unpredictable behavior of the electrode plate above 1 MHz, and temperature measurement showed that heating of the medium influenced the trapping effect, especially for larger amplitudes and higher frequencie

Extracellular stimulation window explained by a geometry-based model of the neuron-electrode contact

Extracellular stimulation of single cultured neurons which are completely sealing a microelectrode is usually performed using anodic or biphasic currents of at least 200 nA. However, recently obtained experimental data demonstrate the possibility to stimulate a neuron using cathodic current pulses with less amplitude. Also, a stimulation window is observed. These findings can be explained by a finite-element model which permits geometry-based electrical representation of the neuron-electrode interface and can be used to explore the required conditions for extracellular stimulation in detail. Modulation of the voltage sensitive channels in the sealing part of the membrane appears to be the key to successful cathodic stimulation. Furthermore, the upper limit of the stimulation window can be explained as a normal consequence of the neuronal membrane electrophysiology

Changes within bursts during learning in dissociated neural networks

We have studied the effect of imprinting a new stimulus-response (SR) relationship into a neuronal network cultured on a multi electrode array (MEA). We have used the Conditional Repetitive Stimulation (CRS) algorithm introduced by Shahaf et al in 2004. In this algorithm focal electrical stimulation is delivered at a low rate (≪1 Hz) and is withdrawn when a desired response is observed. We confirmed that CRS could train the network to strengthen an initially weak SR relationship. With the acquisition of a new SR relationship, we studied its effect on network activity. Specifically, spontaneously occurring network bursts measured before, during and after training were analyzed. The total firing rate within bursts was estimated with a temporal resolution of milliseconds (burst profiles). We have shown earlier that these profiles change shape on a time base of several hours during spontaneous development. We show that the rate of change of the profiles during training (i.e. CRS) was higher than when no stimulation was applied

Geometry-based finite-element modeling of the electrical contact between a cultured neuron and a microelectrode

The electrical contact between a substrate embedded microelectrode and a cultured neuron depends on the geometry of the neuron-electrode interface. Interpretation and improvement of these contacts requires proper modeling of all coupling mechanisms. In literature, it is common practice to model the neuron-electrode contact using lumped circuits in which large simplifications are made in the representation of the interface geometry. In this paper, the finite-element method is used to model the neuron-electrode interface, which permits numerical solutions for a variety of interface geometries. The simulation results offer detailed spatial and temporal information about the combined electrical behavior of extracellular volume, electrode-electrolyte interface and neuronal membrane

Modeled channel distributions explain extracellular recordings from cultured neurons sealed to microelectrodes

Amplitudes and shapes of extracellular recordings from single neurons cultured on a substrate embedded microelectrode depend not only on the volume conducting properties of the neuron-electrode interface, but might also depend on the distribution of voltage-sensitive channels over the neuronal membrane. In this paper, finite-element modeling is used to quantify the effect of these channel distributions on the neuron-electrode contact. Slight accumulation or depletion of voltage-sensitive channels in the sealing membrane of the neuron results in various shapes and amplitudes of simulated extracellular recordings. However, estimation of channel-specific accumulation factors from extracellular recordings can be obstructed by co-occuring ion currents and defect sealing. Experimental data from cultured neuron-electrode interfaces suggest depletion of sodium channels and accumulation of potassium channels

Endoneural selective stimulating using wire-microelectrode arrays

In acute experiments eight 5- to 24-wire-microelectrode arrays were inserted into the common peroneal nerve of the rat, to investigate whether the electrodes could selectively stimulate motor units of the extensor digitorum longus (EDL) muscle. Twitch-force-recruitment curves were measured from the EDL for each array electrode. The curves were plotted on a double-logarithmic scale and parameterized by the low-force slope (which represents the power p in the power-law relationship of force F versus stimulus current I, or F~Ip) and the threshold current. The slopes and threshold currents measured with array electrodes did not differ significantly from those obtained with randomly inserted single wire-microelectrodes. This indicates that, although involving a more invasive insertion procedure, electrode arrays provide neural contacts with low-force recruitment properties similar to those of single wires. Array results revealed partial blocking of neural conduction, similar to that reported with microneurographic insertion with single needles. The efficiency of the array was defined as the fraction of array electrodes selectively contacting a motor unit and evoking the corresponding threshold force. Efficiency thus expresses the practical value of the used electrode array in terms of the total number of distinct threshold forces that can be stimulated by selecting the appropriate electrodes. The eight arrays were capable of evoking threshold forces selectively with an average efficiency of 0.81 (or 81%

Intraneural stimulation using wire-microelectrode arrays: analysis of force steps in recruitment curves

In acute experiments on six Wistar rats, a wire-microelectrode array was inserted into the common peroneal nerve. A 5-channel array and a 24-channel array were available. Each electrode in the array was used to generate a twitch contraction force recruitment curve for the extensor digitorum longus muscle. We constructed a histogram of the pooled force steps in all recruitment curves. From a comparison of this experimental histogram with one estimated from literature data, we found that the force steps encountered in our experiments are in the same range as those from the literature-based estimated distribution. Discrepancies between the experimental and the literature-based histogram might be ascribed to an approximation used in the estimated distribution. We conclude that force step histograms appear to provide a simple means for estimating motor unit twitch force distributions, and thus are of value in studies of intraneural selective stimulatio