Flexible Neural Electrode Array Based-on Porous Graphene for Cortical Microstimulation and Sensing.

Neural sensing and stimulation have been the backbone of neuroscience research, brain-machine interfaces and clinical neuromodulation therapies for decades. To-date, most of the neural stimulation systems have relied on sharp metal microelectrodes with poor electrochemical properties that induce extensive damage to the tissue and significantly degrade the long-term stability of implantable systems. Here, we demonstrate a flexible cortical microelectrode array based on porous graphene, which is capable of efficient electrophysiological sensing and stimulation from the brain surface, without penetrating into the tissue. Porous graphene electrodes show superior impedance and charge injection characteristics making them ideal for high efficiency cortical sensing and stimulation. They exhibit no physical delamination or degradation even after 1 million biphasic stimulation cycles, confirming high endurance. In in vivo experiments with rodents, same array is used to sense brain activity patterns with high spatio-temporal resolution and to control leg muscles with high-precision electrical stimulation from the cortical surface. Flexible porous graphene array offers a minimally invasive but high efficiency neuromodulation scheme with potential applications in cortical mapping, brain-computer interfaces, treatment of neurological disorders, where high resolution and simultaneous recording and stimulation of neural activity are crucial

Design, fabrication, and test of a sensor array for perspective biosensing in chronic pathologies

Biosensing for chronic pathologies requires the simultaneous monitoring of different parameters such as drug concentrations, inflammation status, temperature and pH. In this paper we discuss the design, fabrication and test of a sensor array hosting five biosensor platforms, a pH electrode and a temperature sensor. Different and reproducible nano-bio- functionalization can be obtained with high spatial resolution via selective electrodeposition of chitosan/MWCNT/enzyme solutions at the various electrodes. The array, completely fabricated with biocompatible materials, can be integrated with a CMOS integrated circuit and a remote powering coil for the realization of a fully implantable device

Poly(3,4-ethylenedioxythiophene) as a Micro-Neural Interface Material for Electrostimulation

Chronic microstimulation-based devices are being investigated to treat conditions such as blindness, deafness, pain, paralysis, and epilepsy. Small-area electrodes are desired to achieve high selectivity. However, a major trade-off with electrode miniaturization is an increase in impedance and charge density requirements. Thus, the development of novel materials with lower interfacial impedance and enhanced charge storage capacity is essential for the development of micro-neural interface-based neuroprostheses. In this report, we study the use of conducting polymer poly(3,4-ethylenedioxythiophene) (PEDOT) as a neural interface material for microstimulation of small-area iridium electrodes on silicon-substrate arrays. Characterized by electrochemical impedance spectroscopy, electrodeposition of PEDOT results in lower interfacial impedance at physiologically relevant frequencies, with the 1 kHz impedance magnitude being 23.3 ± 0.7 kΩ, compared to 113.6 ± 3.5 kΩ for iridium oxide (IrOx) on 177 μm2 sites. Further, PEDOT exhibits enhanced charge storage capacity at 75.6 ± 5.4 mC/cm2 compared to 28.8 ± 0.3 mC/cm2 for IrOx, characterized by cyclic voltammetry (50 mV/s). These improvements at the electrode interface were corroborated by observation of the voltage excursions that result from constant current pulsing. The PEDOT coatings provide both a lower amplitude voltage and a more ohmic representation of the applied current compared to IrOx. During repetitive pulsing, PEDOT-coated electrodes show stable performance and little change in electrical properties, even at relatively high current densities which cause IrOx instability. These findings support the potential of PEDOT coatings as a micro-neural interface material for electrostimulation

Iridium Oxide Microelectrode Arrays for In Vitro Stimulation of Individual Rat Neurons from Dissociated Cultures

We present the first in vitro extracellular stimulation of individual neurons from dissociated cultures with iridium oxide (IrOx) electrodes. Microelectrode arrays with sputtered IrOx films (SIROF) were developed for electrophysiological investigations with electrogenic cells. The microelectrodes were characterized with scanning electron and atomic force microscopy, revealing rough and porous electrodes with enlarged surface areas. As shown by cyclic voltammetry and electrochemical impedance spectroscopy, the large surface area in combination with the good electrochemical properties of SIROF resulted in high charge storage capacity and low electrode impedance. Thus, we could transfer the good properties of IrOx as material for in vivo stimulation electrodes to multi-electrode arrays with electrode diameters as small as 10 μm for in vitro applications. Single rat cortical neurons from dissociated cultures were successfully stimulated to fire action potentials using single or trains of biphasic rectangular voltage-controlled stimulation pulses. The stimulated cell's membrane potential was simultaneously monitored using whole-cell current-clamp recordings. This experimental configuration allowed direct evaluation of the influence of pulse phase sequence, amplitude, and number on the stimulation success ratio and action potential latency. Negative phase first pulses were more effective for extracellular stimulation and caused reduced latency in comparison to positive phase first pulses. Increasing the pulse amplitude also improved stimulation reliability. However, in order to prevent cell or electrode damage, the pulse amplitude is limited to voltages below the threshold for irreversible electrochemical reactions at the electrode. As an alternative to increasing the amplitude, a higher number of stimulation pulses was also shown to increase stimulation success

Ion Beam Assisted E-Beam Deposited TiN Microelectrodes—Applied to Neuronal Cell Culture Medium Evaluation

Microelectrode material and cell culture medium have significant roles in the signal-to-noise ratio and cell well-being in in vitro electrophysiological studies. Here, we report an ion beam assisted e-beam deposition (IBAD) based process as an alternative titanium nitride (TiN) deposition method for sputtering in the fabrication of state-of-the-art TiN microelectrode arrays (MEAs). The effects of evaporation and nitrogen flow rates were evaluated while developing the IBAD TiN deposition process. Moreover, the produced IBAD TiN microelectrodes were characterized by impedance, charge transfer capacity (CTC) and noise measurements for electrical properties, AFM and SEM for topological imaging, and EDS for material composition. The impedance (at 1 kHz) of brand new 30 μm IBAD TiN microelectrodes was found to be double but still below 100 kΩ compared with commercial reference MEAs with sputtered TiN microelectrodes of the same size. On the contrary, the noise level of IBAD TiN MEAs was lower compared with that of commercial sputtered TiN MEAs in equal conditions. In CTC IBAD TiN electrodes (3.3 mC/cm2) also outperformed the sputtered counterparts (2.0 mC/cm2). To verify the suitability of IBAD TiN microelectrodes for cell measurements, human pluripotent stem cell (hPSC)-derived neuronal networks were cultured on IBAD TiN MEAs and commercial sputtered TiN MEAs in two different media: neural differentiation medium (NDM) and BrainPhys (BPH). The effect of cell culture media to hPSC derived neuronal networks was evaluated to gain more stable and more active networks. Higher spontaneous activity levels were measured from the neuronal networks cultured in BPH compared with those in NDM in both MEA types. However, BPH caused more problems in cell survival in long-term cultures by inducing neuronal network retraction and clump formation after 1–2 weeks. In addition, BPH was found to corrode the Si3N4 insulator layer more than NDM medium. The developed IBAD TiN process gives MEA manufacturers more choices to choose which method to use to deposit TiN electrodes and the medium evaluation results remind that not only electrode material but also insulator layer and cell culturing medium have crucial role in successful long term MEA measurements

Single-Cell Electrical Stimulation Using CMOS-Based High-Density Microelectrode Arrays

Non-invasive electrical stimulation can be used to study and control neural activity in the brain or to alleviate somatosensory dysfunctions. One intriguing prospect is to precisely stimulate individual targeted neurons. Here, we investigated single-neuron current and voltage stimulation in vitro using high-density microelectrode arrays featuring 26,400 bidirectional electrodes at a pitch of 17.5 μm and an electrode area of 5 × 9 μm2. We determined optimal waveforms, amplitudes and durations for both stimulation modes. Owing to the high spatial resolution of our arrays and the close proximity of the electrodes to the respective neurons, we were able to stimulate the axon initial segments (AIS) with charges of less than 2 pC. This resulted in minimal artifact production and reliable readout of stimulation efficiency directly at the soma of the stimulated cell. Stimulation signals as low as 70 mV or 100 nA, with pulse durations as short as 18 μs, yielded measurable action potential initiation and propagation. We found that the required stimulation signal amplitudes decreased with cell growth and development and that stimulation efficiency did not improve at higher electric fields generated by simultaneous multi-electrode stimulation

Metallized printed microstructures for precision biomedical recording and stimulation

Implantable electrodes are the central tool for many techniques and treatments in biomedical research and medicine. There is a trend in these tools towards arrays of tissue-penetrating microelectrodes with low geometric surface areas for purposes of both increasing the specificity of recording/stimulation and reducing tissue damage due to insertion trauma and reactive immune responses. However, smaller electrode sizes present new constraints – both difficulty in fabrication as well as significant limitations on effective charge storage/injection capacities as well as higher impedances, making smaller electrodes less capable of easily passing charge safely and efficiently. Fabricating structures on the scale of tens of microns and below poses significant challenges compared to well established machining at larger sizes. Established sets of techniques such as classic MEMS processes are limited to relatively specific shapes, with significant limitations in their ability to produce curved surfaces and surfaces which are not composed of highly distinct stepped layers. We developed a method for improvement of impedance and charge storage capacity of flat electrodes without affecting geometric surface area (footprint) using Resonant Direct Laser Writing (rDLW) 3D printing to fabricate high surface area 3D structures, which were then rendered conductive. The ability to perform DLW printing at a range of laser powers on opaque reflective surfaces is demonstrated, previously a known limitation of direct laser writing. This is demonstrated through a variety of example prints. This capability opens the door to many new possibilities in micron resolution polymer printing which were previously inaccessible, with potentially far reaching ramifications for microfabrication

In vitro feasibility testing of floating light-activated minroelectrical stimulators

One of the major challenges of neural stimulation is the mechanical stress and resulting trauma induced on the implanted electrodes by the constant movement of the interconnects. A potential way of eliminating interconnects is to use floating micro-stimulators that can be activated through optical means. As a method of energy transfer to the micro-stimulator, we propose to use a laser beam at near infrared (NIR) wavelengths. There are two main objectives in this project to test the feasibility of the main approach; investigate the charge injection capacity of titanium nitride (TiN) and iridium oxide (IrOx) as potential contact materials, and measure the transmitted light power through the neural tissue for various implantation depths. The charge injection capacity of TiN electrodes for an extended range of cathodic voltages was also investigated. Because the microstimulator will be implanted into the neural tissue, the laser beam must penetrate a few millimeters before reaching the device. The transmitted light power was measured for various types of neural tissue. The transmitted light power through rat brain gray matter was much higher than that of the white matter and the sciatic nerve. Penetration depth and reflectance were calculated according to Lambert-Beer’s law from measurements of transmission for various tissue thicknesses. The results suggest that FLAMES approach is feasible for implantation depths of a few millimeters in the peripheral and central nervous system. Both IrOx and TiN allow sufficient charge injection for this application. TiN is preferred for future experimentation since TiN does not require a bias voltage to achieve useful charge injection rates, and thus is a good choice as an electrode material in this application

Direct growth of carbon nanotubes on new high-density 3D pyramid-shaped microelectrode arrays for brain-machine interfaces

Silicon micromachined, high-density, pyramid-shaped neural microelectrode arrays (MEAs) have been designed and fabricated for intracortical 3D recording and stimulation. The novel architecture of this MEA has made it unique among the currently available micromachined electrode arrays, as it has provided higher density contacts between the electrodes and targeted neural tissue facilitating recording from different depths of the brain. Our novel masking technique enhances uniform tip-exposure for variable-height electrodes and improves process time and cost significantly. The tips of the electrodes have been coated with platinum (Pt). We have reported for the first time a selective direct growth of carbon nanotubes (CNTs) on the tips of 3D MEAs using the Pt coating as a catalyzer. The average impedance of the CNT-coated electrodes at 1 kHz is 14 k. The CNT coating led to a 5-fold decrease of the impedance and a 600-fold increase in charge transfer compared with the Pt electrode