Extracellular Recording System Research Articles

The response of the neuronal activity in the somatosensory cortex after high-intensity intermediate-frequency magnetic field exposure to the spinal cord in rats under anesthesia and waking states

Novel technologies using the intermediate-frequency magnetic field (IF-MF) in living environments are becoming popular with the advance in electricity utilization. However, the biological effects induced by the high-intensity and burst-type IF-MF exposure used in the wireless power transfer technologies for electric vehicles or medical devices, such as the magnetic stimulation techniques, are not well understood. Here, we developed an experimental platform using rats, that combined an 18 kHz, high-intensity (Max. 88 mT), Gaussian-shaped burst IF-MF exposure system with an in vivo extracellular recording system. In this paper, we aimed to report the qualitative differences in stimulus responses in the regions of the somatosensory cortex and peripheral nerve fibers that were induced by the IF-MF exposure to the rat spinal cord. We also report the modulation of the stimulus responses in the somatosensory cortex under anesthesia or waking states. Using this experimental platform, we succeeded in the detection of the motor evoked potentials or the neuronal activity in the somatosensory cortex that was induced by the IF-MF exposure to the spinal cord in rats. Compared to the state of anesthesia, the neuronal activities in the somatosensory cortex was enhanced during the waking state. On the other hand, these neuronal responses could not be confirmed by the IF-MF exposure-related coil sound only. Our experimental results indicated the basic knowledge of the biological responses and excitation mechanisms of the spinal cord stimulation by the IF-MF exposure.

A Novel Algorithm for Online Spike Detection

Recordings of extracellular spikes have been widely used in various fields ranging from basic neuroscientific research to clinical applications. However, in the extracellular recording system, how to accurately detect spikes from the recorded signal in real time is still a major challenging work. Although the existing algorithms for online spike detection have made great progress, there still remains much room for improvement in terms of accuracy. In this paper, we propose a new method for high accuracy and real time spike detection. Concretely, differential operator is firstly employed to accentuate spikes in the signal for its simplicity and strong capacity to detect significant changes. Then, by exploiting the structural features of spikes, the resolution parameter is introduced to improve the performance of differential operator. Finally, a simple and effective measure is utilized to further reduce the influence of background noise, which makes spike detection more accurate. The results of simulated and real data show that the proposed method is able to precisely detect spikes while maintaining low computational complexity.

Optimization and pharmacological characterization of iCell cardiomyocytes for electrophysiological assessment in a multi-electrode extracellular recording system

Optimization and pharmacological characterization of iCell cardiomyocytes for electrophysiological assessment in a multi-electrode extracellular recording system

Electrical interaction between cardiomyocyte sheets separated by non-cardiomyocyte sheets in heterogeneous tissues

Electrical coupling between cardiomyocytes is important in synchronous beating and normal heart functions. Cardiomyocytes are also electrically coupled to non-cardiomyocytes. The electrical interactions between cardiomyocytes and non-cardiomyocytes, or those between separated cardiomyocytes, are important for normal heart function because abnormalities of the coupling and variation of the cell population induce pathological heart functions and arrhythmias. In this study the three-dimensional time course of the electrical interaction between two rat neonatal cardiomyocyte sheets separated by non-cardiomyocyte sheets was analysed by a multiple-electrode extracellular recording system. The two cardiomyocyte sheets separated by a single- or double-layered mouse fibroblast NIH3T3 cell sheet coupled electrically at 113 +/- 28 or 287 +/- 87 min after layering, respectively. The time course of the electrical coupling, when the single-layer NIH3T3 cell sheet was inserted, is similar to that of a layered cardiomyocyte sheet. Immunocytological analysis and dye transfer assay suggested the formation of gap junctions at heterocellular junctions of cardiomyocytes and NIH3T3 cells. On the other hand, when a double-layered NIH3T3 cell sheet was inserted, an incomplete electrical coupling of two cardiomyocyte sheets, including a conduction delay, was observed. The electrical coupling of cardiomyocyte sheets was completely blocked (conduction block) by insertion of a triple-layered NIH3T3 cell sheet, a communication-defective HeLa cell sheet or a Ca(2+)-antagonist LaCl(3)-treated cell sheet. These electrophysiological analyses of heterogeneously stacked cell sheets might provide insights into complex electrical conduction systems that resemble those of native or damaged heart and transplanted tissues.

Electrical coupling of cardiomyocyte sheets occurs rapidly via functional gap junction formation

Previously, we have successfully created pulsatile myocardial tissue grafts using our novel technology, "cell sheet engineering", that layers cell sheets fabricated on temperature-responsive culture dishes to form three-dimensional (3-D) structures. Electrical coupling is established between layered neonatal rat cardiomyocyte sheets, resulting in the synchronized beating of 3-D myocardial tissues. However, the mechanism by which these layered cardiomyocyte sheets communicate electrically is not well-understood. In this study, we used a multiple-electrode extracellular recording system and demonstrated that bilayer cardiomyocyte sheets coupled electrically with slight delays 34+/-2 min (mean+/-SEM) after layering. These delays gradually decreased and the electrical actions of layered cell sheets were completely coupled 46+/-3 min (mean+/-SEM) after initial layering. Immunohistological analysis showed that connexin43, a gap junction (GJ)-related protein, existed not only at cell-to-cell interfaces but also on the free cell membrane in the cardiomyocyte sheet. Additionally, neither connexin40 nor connexin45, but only connexin43 was detected between bilayer cardiomyocyte sheets within 30 min after layering. Dye transfer assay demonstrated that the exchange of small molecules via GJs occurred within 30 min. The cell sheet manipulation technique using the temperature-responsive culture dishes has substantial advances and the exciting potential in the fields of cell and tissue physiology, as well as tissue engineering.

64-Channel extended gate electrode arrays for extracellular signal recording

A 64-channel amplifier system for the recording of extracellular signals with planar metal microelectrodes is presented. Gold metal microelectrodes on glass wafers were fabricated using standard photolithographic techniques. The measurement system was divided into a headstage preamplifier and a main amplifier. The inherent noise of the extracellular recording system was minimized by using an independent battery supply. The metal electrodes were directly connected to the gates of low noise junction field effect transistors (JFETs) using a specially designed electronic circuit. With this set-up, it was possible to record extracellular signals with planar metal microelectrodes without any surface modification for impedance reduction. A feedback circuit in the first amplification stage compensated slow drifts of the gold microelectrodes, which made online sampling of all 64 channels with a sampling rate of 10 kHz possible. Recordings were taken from rat cardiac myocytes cultured on fibronectin coated sensor chips. The system exhibited a good signal-to-noise ratio. It was able to detect the signal propagation within the cardiac cell layer and it could be used for pharmacological investigations involving the heart.

Cardiomyocyte-transistor-hybrids for sensor application

An extracellular recording system has been designed for the detection of electrical cell signals using p-channel or n-channel field-effect transistor (FET) arrays with non-metallized gates. Signals from rat heart muscle cell were recorded by these devices and the results described on the basis of an equivalent circuit. This technique is sensitive enough to detect minute changes of the extracellular membrane voltage and has potential applications in drug screening. We show that known cardiac stimulants (isoproterenol, norepinephrine) and relaxants (verapamil, carbamylcholine) have characteristic effects on the heart cells in terms of the changes of beat frequencies in the absence or presence of corresponding agents.

Polysynaptic excitatory pathways induce heterosynaptic depression in the rat auditory cortex.

Short-term plasticity, the effect of a preceding synaptic response on the following response in a pair, in layers II/III of the rat auditory cortex slice after application of repetitive stimuli to layer IV, was investigated using a multichannel extracellular recording system. Paired-pulse depression, which is induced to a moderate degree in standard artificial cerebrospinal fluid, was markedly facilitated in the presence of bicuculline, a GABA(A) receptor antagonist, concurrent with the emergence of a polysynaptic component of the EPSP (polyEPSP) in the first response in a pair. This depression (bicuculline-facilitated synaptic depression, BFSD) was maximal at the minimum interval tested (50 ms), reduced as the interval was increased, and persisted beyond an interval of 2 s. The occurrence of BFSD was dependent on the presence of a polyEPSP regardless of the presence of the monosynaptic component of the EPSP, indicating that BFSD is induced by a heterosynaptic mechanism. D-AP5, an NMDA receptor antagonist, partially eliminated polyEPSPs and reversed BFSD. These results suggest that activation of polysynaptic excitatory pathways induces a heterosynaptic depression in the range of a few seconds and that NMDA receptor activity is involved in this heterosynaptic depression.

Description and demonstration of a CMOS amplifier-based-system with measurement and stimulation capability for bioelectrical signal transduction

An extracellular recording system incorporating an electrode array and an amplifier/stimulator CMOS chip is described and characterized. Important features of this custom VLSI chip include 16 instrumentation amplifiers with a gain of 50 and the incorporation of a cross-point array allowing designation of an extracellular microelectrode as either a stimulator or sensor. The planar array consisted of 32 microelectrodes, 14 μm in diameter, and four larger reference electrodes. Microelectrodes, interconnecting traces, and bond pads were patterned with a 500-nm layer of gold. The interconnecting traces were passivated with a 1-μm thick layer of silicon nitride to provide chemical and electrical insulation and microelectrode impedance was lowered utilizing electrode position of platinum black. The amplifier exhibited a nearly flat frequency response with high pass and low pass corner frequencies of 0.7 Hz and 50 kHz, respectively. The input referred noise over the 50 kHz bandwidth was 12–16 μV RMS, well below the magnitude of previously reported extracellular potentials. Crosstalk between neighboring channels resulted in an output signal below the amplifier noise level, even for relatively large extracellular potentials. Using this system, extracellular recordings were demonstrated yielding typical peak-to-peak biopotentials of magnitude 0.9–2.1 mV and 100–400 μV for chick cardiac myocytes and rat spinal cord neurons, respectively. The key components of this extracellular recording system can be manufactured using industry standard thin film photolithographic techniques.