An electroencephalogram microdisplay to visualize neuronal activity on the brain surface.

Functional mapping during brain surgery is applied to define brain areas that control critical functions and cannot be removed. Currently, these procedures rely on verbal interactions between the neurosurgeon and electrophysiologist, which can be time-consuming. In addition, the electrode grids that are used to measure brain activity and to identify the boundaries of pathological versus functional brain regions have low resolution and limited conformity to the brain surface. Here, we present the development of an intracranial electroencephalogram (iEEG)-microdisplay that consists of freestanding arrays of 2048 GaN light-emitting diodes laminated on the back of micro-electrocorticography electrode grids. With a series of proof-of-concept experiments in rats and pigs, we demonstrate that these iEEG-microdisplays allowed us to perform real-time iEEG recordings and display cortical activities by spatially corresponding light patterns on the surface of the brain in the surgical field. Furthermore, iEEG-microdisplays allowed us to identify and display cortical landmarks and pathological activities from rat and pig models. Using a dual-color iEEG-microdisplay, we demonstrated coregistration of the functional cortical boundaries with one color and displayed the evolution of electrical potentials associated with epileptiform activity with another color. The iEEG-microdisplay holds promise to facilitate monitoring of pathological brain activity in clinical settings.

A 0.00179 mm2/Ch Chopper-Stabilized TDMA Neural Recording System with Dynamic EOV Cancellation and Predictive Mixed-Signal Impedance Boosting.

This article presents a digitally-assisted multi-channel neural recording system. The system uses a 16-channel chopper-stabilized Time Division Multiple Access (TDMA) scheme to record multiplexed neural signals into a single shared analog front end (AFE). The choppers reduce the total integrated noise across the modulated spectrum by 2.4× and 4.3× in Local Field Potential (LFP) and Action Potential (AP) bands, respectively. In addition, a novel impedance booster based on Sign-Sign least mean squares (LMS) adaptive filter (AF) predicts the input signal and pre-charges the AC-coupling capacitors. The impedance booster module increases the AFE input impedance by a factor of 39× with a 7.1% increase in area. The proposed system obviates the need for on-chip digital demodulation, filtering, and remodulation normally required to extract Electrode Offset Voltages (EOV) from multiplexed neural signals, thereby achieving 3.6× and 2.8× savings in both area and power, respectively, in the EOV filter module. The Sign-Sign LMS AF is reused to determine the system loop gain, which relaxes the feedback DAC accuracy requirements and saves 10.1× in power compared to conventional oversampled DAC truncation-error ΔΣ-modulator. The proposed SoC is designed and fabricated in 65 nm CMOS, and each channel occupies 0.00179 mm2 of active area. Each channel consumes 5.11 µW of power while achieving 2.19 µVrms and 2.4 µVrms of input referred noise (IRN) over AP and LFP bands. The resulting AP band noise efficiency factor (NEF) is 1.8. The proposed system is verified with acute in-vivo recordings in a Sprague-Dawley rat using parylene C based thin-film platinum nanorod microelectrodes.

Flexible, scalable, high channel count stereo-electrode for recording in the human brain.

Over the past decade, stereotactically placed electrodes have become the gold standard for deep brain recording and stimulation for a wide variety of neurological and psychiatric diseases. Current electrodes, however, are limited in their spatial resolution and ability to record from small populations of neurons, let alone individual neurons. Here, we report on an innovative, customizable, monolithically integrated human-grade flexible depth electrode capable of recording from up to 128 channels and able to record at a depth of 10 cm in brain tissue. This thin, stylet-guided depth electrode is capable of recording local field potentials and single unit neuronal activity (action potentials), validated across species. This device represents an advance in manufacturing and design approaches which extends the capabilities of a mainstay technology in clinical neurology.

Modular Phoneme Processing in Human Superior Temporal Gyrus.

Modular organization is fundamental to cortical processing, but its presence is human association cortex is unknown. We characterized phoneme processing with 128-1024 channel micro-arrays at 50-200µm pitch on superior temporal gyrus of 7 patients. High gamma responses were highly correlated within ~1.7mm diameter modules, sharply delineated from adjacent modules with distinct time-courses and phoneme-selectivity. We suggest that receptive language cortex may be organized in discrete processing modules.

Intraoperative application and early experience with novel high-resolution, high-channel-count thin-film electrodes for human microelectrocorticography.

OBJECTIVE: The study objective was to evaluate intraoperative experience with newly developed high-spatial-resolution microelectrode grids composed of poly(3,4-ethylenedioxythiophene) with polystyrene sulfonate (PEDOT:PSS), and those composed of platinum nanorods (PtNRs). METHODS: A cohort of patients who underwent craniotomy for pathological tissue resection and who had high-spatial-resolution microelectrode grids placed intraoperatively were evaluated. Patient demographic and baseline clinical variables as well as relevant microelectrode grid characteristic data were collected. The primary and secondary outcome measures of interest were successful microelectrode grid utilization with usable resting-state or task-related data, and grid-related adverse intraoperative events and/or grid dysfunction. RESULTS: Included in the analysis were 89 cases of patients who underwent a craniotomy for resection of neoplasms (n = 58) or epileptogenic tissue (n = 31). These cases accounted for 94 grids: 58 PEDOT:PSS and 36 PtNR grids. Of these 94 grids, 86 were functional and used successfully to obtain cortical recordings from 82 patients. The mean cortical grid recording duration was 15.3 ± 1.15 minutes. Most recordings in patients were obtained during experimental tasks (n = 52, 58.4%), involving language and sensorimotor testing paradigms, or were obtained passively during resting state (n = 32, 36.0%). There were no intraoperative adverse events related to grid placement. However, there were instances of PtNR grid dysfunction (n = 8) related to damage incurred by suboptimal preoperative sterilization (n = 7) and improper handling (n = 1); intraoperative recordings were not performed. Vaporized peroxide sterilization was the most optimal sterilization method for PtNR grids, providing a significantly greater number of usable channels poststerilization than did steam-based sterilization techniques (median 905.0 [IQR 650.8-935.5] vs 356.0 [IQR 18.0-597.8], p = 0.0031). CONCLUSIONS: High-spatial-resolution microelectrode grids can be readily incorporated into appropriately selected craniotomy cases for clinical and research purposes. Grids are reliable when preoperative handling and sterilization considerations are accounted for. Future investigations should compare the diagnostic utility of these high-resolution grids to commercially available counterparts and assess whether diagnostic discrepancies relate to clinical outcomes.

Synthesis of PEDOT:PSS Brushes Grafted from Gold Using ATRP for Increased Electrochemical and Mechanical Stability.

We report PEDOT:PSS brushes grafted from gold using surface-initiated atom-transfer radical polymerization (SI-ATRP) which demonstrate significantly enhanced mechanical stability against sonication and electrochemical cycling compared to spin-coated analogues as well as lower impedances than bare gold at frequencies from 0.1 to 105 Hz. These results suggest SI-ATRP PEDOT:PSS to be a promising candidate for use in microelectrodes for neural activity recording. Spin-coated, electrodeposited, and drop-cast PEDOT:PSS have already been shown to reduce impedance and improve biocompatibility of microelectrodes, but the lack of strong chemical bonds of the physisorbed polymer film to the metal leads to disintegration under required operational stresses including cyclic mechanical loads, abrasion, and electrochemical cycling. Rather than modifying the metal electrode or introducing cross-linkers or other additives to improve the stability of the polymer film, this work chemically tethers the polymer to the surface, offering a simple, scalable solution for functional bioelectronic interfaces.

The Ten Commandments of Everyday Leadership.

The editorial offers principles of everyday leadership, with hope for the new year.

Conductive electrospun polymer improves stem cell-derived cardiomyocyte function and maturation.

Despite numerous efforts to generate mature human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs), cells often remain immature, electrically isolated, and may not reflect adult biology. Conductive polymers are attractive candidates to facilitate electrical communication between hPSC-CMs, especially at sub-confluent cell densities or diseased cells lacking cell-cell junctions. Here we electrospun conductive polymers to create a conductive fiber mesh and assess if electrical signal propagation is improved in hPSC-CMs seeded on the mesh network. Matrix characterization indicated fiber structure remained stable over weeks in buffer, scaffold stiffness remained near in vivo cardiac stiffness, and electrical conductivity scaled with conductive polymer concentration. Cells remained adherent and viable on the scaffolds for at least 5 days. Transcriptomic profiling of hPSC-CMs cultured on conductive substrates for 3 days showed upregulation of cardiac and muscle-related genes versus non-conductive fibers. Structural proteins were more organized and calcium handling was improved on conductive substrates, even at sub-confluent cell densities; prolonged culture on conductive scaffolds improved membrane depolarization compared to non-conductive substrates. Taken together, these data suggest that blended, conductive scaffolds are stable, supportive of electrical coupling in hPSC-CMs, and promote maturation, which may improve our ability to model cardiac diseases and develop targeted therapies.

A Comprehensive Large Signal, Small Signal, and Noise Model for IGZO Thin Film Transistor Circuits.

We report a new physics-based model for dual-gate amorphous-indium gallium zinc oxide (a-IGZO) thin film transistors (TFTs) which we developed and fine-tuned through experimental implementation and benchtop characterization. We fabricated and characterized a variety of test patterns, including a-IGZO TFTs with varying gate widths (100-1000 µm) and channel lengths (5-50 µm), transmission-line-measurement patterns and ground-signal-ground (GSG) radio frequency (RF) patterns. We modeled the contact resistance as a function of bias, channel area, and temperature, and captured all operating regimes, used physics-based modeling adjusted for empirical data to capture the TFT characteristics including ambipolar subthreshold currents, graded interbias-regime current changes, threshold and flat-band voltages, the interface trap density, the gate leakage currents, the noise, and the relevant small signal parameters. To design high-precision circuits for biosensing, we validated the dc, small signal, and noise characteristics of the model. We simulated and fabricated a two-stage common source amplifier circuit with a common drain output buffer and compared the measured and simulated gain and phase performance, finding an excellent fit over a frequency range spanning 10 kHz-10 MHz.

The Brain Electroencephalogram Microdisplay for Precision Neurosurgery.

Brain surgeries are among the most delicate clinical procedures and must be performed with the most technologically robust and advanced tools. When such surgical procedures are performed in functionally critical regions of the brain, functional mapping is applied as a standard practice that involves direct coordinated interactions between the neurosurgeon and the clinical neurology electrophysiology team. However, information flow during these interactions is commonly verbal as well as time consuming which in turn increases the duration and cost of the surgery, possibly compromising the patient outcomes. Additionally, the grids that measure brain activity and identify the boundaries of pathological versus functional brain regions suffer from low resolution (3-10 mm contact to contact spacing) with limited conformity to the brain surface. Here, we introduce a brain intracranial electroencephalogram microdisplay (Brain-iEEG-microdisplay) which conforms to the brain to measure the brain activity and display changes in near real-time (40 Hz refresh rate) on the surface of the brain in the surgical field. We used scalable engineered gallium nitride (GaN) substrates with 6" diameter to fabricate, encapsulate, and release free-standing arrays of up to 2048 GaN light emitting diodes (µLEDs) in polyimide substrates. We then laminated the µLED arrays on the back of micro-electrocorticography (µECoG) platinum nanorod grids (PtNRGrids) and developed hardware and software to perform near real-time intracranial EEG analysis and activation of light patterns that correspond to specific cortical activities. Using the Brain-iEEG-microdisplay, we precisely ideFSntified and displayed important cortical landmarks and pharmacologically induced pathological activities. In the rat model, we identified and displayed individual cortical columns corresponding to individual whiskers and the near real-time evolution of epileptic discharges. In the pig animal model, we demonstrated near real-time mapping and display of cortical functional boundaries using somatosensory evoked potentials (SSEP) and display of responses to direct electrical stimulation (DES) from the surface or within the brain tissue. Using a dual-color Brain-iEEG-microdisplay, we demonstrated co-registration of the functional cortical boundaries with one color and displayed the evolution of electrical potentials associated with epileptiform activity with another color. The Brain-iEEG-microdisplay holds the promise of increasing the efficiency of diagnosis and possibly surgical treatment, thereby reducing the cost and improving patient outcomes which would mark a major advancement in neurosurgery. These advances can also be translated to broader applications in neuro-oncology and neurophysiology.

Electrochemical and electrophysiological considerations for clinical high channel count neural interfaces.

Electrophysiological recording and stimulation are the gold standard for functional mapping during surgical and therapeutic interventions as well as capturing cellular activity in the intact human brain. A critical component probing human brain activity is the interface material at the electrode contact that electrochemically transduces brain signals to and from free charge carriers in the measurement system. Here, we summarize state-of-the-art electrode array systems in the context of translation for use in recording and stimulating human brain activity. We leverage parametric studies with multiple electrode materials to shed light on the varied levels of suitability to enable high signal-to-noise electrophysiological recordings as well as safe electrophysiological stimulation delivery. We discuss the effects of electrode scaling for recording and stimulation in pursuit of high spatial resolution, channel count electrode interfaces, delineating the electrode-tissue circuit components that dictate the electrode performance. Finally, we summarize recent efforts in the connectorization and packaging for high channel count electrode arrays and provide a brief account of efforts toward wireless neuronal monitoring systems.

Scalable Thousand Channel Penetrating Microneedle Arrays on Flex for Multimodal and Large Area Coverage BrainMachine Interfaces.

The Utah array powers cutting-edge projects for restoration of neurological function, such as BrainGate, but the underlying electrode technology has itself advanced little in the last three decades. Here, advanced dual-side lithographic microfabrication processes is exploited to demonstrate a 1024-channel penetrating silicon microneedle array (SiMNA) that is scalable in its recording capabilities and cortical coverage and is suitable for clinical translation. The SiMNA is the first penetrating microneedle array with a flexible backing that affords compliancy to brain movements. In addition, the SiMNA is optically transparent permitting simultaneous optical and electrophysiological interrogation of neuronal activity. The SiMNA is used to demonstrate reliable recordings of spontaneous and evoked field potentials and of single unit activity in chronically implanted mice for up to 196 days in response to optogenetic and to whisker air-puff stimuli. Significantly, the 1024-channel SiMNA establishes detailed spatiotemporal mapping of broadband brain activity in rats. This novel scalable and biocompatible SiMNA with its multimodal capability and sensitivity to broadband brain activity will accelerate the progress in fundamental neurophysiological investigations and establishes a new milestone for penetrating and large area coverage microelectrode arrays for brain-machine interfaces.

A universal model of electrochemical safety limits in vivo for electrophysiological stimulation.

Electrophysiological stimulation has been widely adopted for clinical diagnostic and therapeutic treatments for modulation of neuronal activity. Safety is a primary concern in an interventional design leveraging the effects of electrical charge injection into tissue in the proximity of target neurons. While modalities of tissue damage during stimulation have been extensively investigated for specific electrode geometries and stimulation paradigms, a comprehensive model that can predict the electrochemical safety limits in vivo doesn't yet exist. Here we develop a model that accounts for the electrode geometry, inter-electrode separation, material, and stimulation paradigm in predicting safe current injection limits. We performed a parametric investigation of the stimulation limits in both benchtop and in vivo setups for flexible microelectrode arrays with low impedance, high geometric surface area platinum nanorods and PEDOT:PSS, and higher impedance, planar platinum contacts. We benchmark our findings against standard clinical electrocorticography and depth electrodes. Using four, three and two contact electrochemical impedance measurements and comprehensive circuit models derived from these measurements, we developed a more accurate, clinically relevant and predictive model for the electrochemical interface potential. For each electrode configuration, we experimentally determined the geometric correction factors that dictate geometry-enforced current spreading effects. We also determined the electrolysis window from cyclic-voltammetry measurements which allowed us to calculate stimulation current safety limits from voltage transient measurements. From parametric benchtop electrochemical measurements and analyses for different electrode types, we created a predictive equation for the cathodal excitation measured at the electrode interface as a function of the electrode dimensions, geometric factor, material and stimulation paradigm. We validated the accuracy of our equation in vivo and compared the experimentally determined safety limits to clinically used stimulation protocols. Our new model overcomes the design limitations of Shannon's equation and applies to macro- and micro-electrodes at different density or separation of contacts, captures the breakdown of charge-density based approaches at long stimulation pulse widths, and invokes appropriate power exponents to current, pulse width, and material/electrode-dependent impedance.

Constructing 2D maps of human spinal cord activity and isolating the functional midline with high-density microelectrode arrays.

Intraoperative neuromonitoring (IONM) is a widely used practice in spine surgery for early detection and minimization of neurological injury. IONM is most commonly conducted by indirectly recording motor and somatosensory evoked potentials from either muscles or the scalp, which requires large-amplitude electrical stimulation and provides limited spatiotemporal information. IONM may inform of inadvertent events during neurosurgery after they occur, but it does not guide safe surgical procedures when the anatomy of the diseased spinal cord is distorted. To overcome these limitations and to increase our understanding of human spinal cord neurophysiology, we applied a microelectrode array with hundreds of channels to the exposed spinal cord during surgery and resolved spatiotemporal dynamics with high definition. We used this method to construct two-dimensional maps of responsive channels and define with submillimeter precision the electrophysiological midline of the spinal cord. The high sensitivity of our microelectrode array allowed us to record both epidural and subdural responses at stimulation currents that are well below those used clinically and to resolve postoperative evoked potentials when IONM could not. Together, these advances highlight the potential of our microelectrode arrays to capture previously unexplored spinal cord neural activity and its spatiotemporal dynamics at high resolution, offering better electrophysiological markers that can transform IONM.

Ultra-Sharp Nanowire Arrays Natively Permeate, Record, and Stimulate Intracellular Activity in Neuronal and Cardiac Networks.

We report innovative scalable, vertical, ultra-sharp nanowire arrays that are individually addressable to enable long-term, native recordings of intracellular potentials. Stable amplitudes of intracellular potentials from 3D tissue-like networks of neurons and cardiomyocytes are obtained. Individual electrical addressability is necessary for high-fidelity intracellular electrophysiological recordings. This study paves the way toward predictive, high-throughput, and low-cost electrophysiological drug screening platforms.

Human brain mapping with multithousand-channel PtNRGrids resolves spatiotemporal dynamics.

Electrophysiological devices are critical for mapping eloquent and diseased brain regions and for therapeutic neuromodulation in clinical settings and are extensively used for research in brain-machine interfaces. However, the existing clinical and experimental devices are often limited in either spatial resolution or cortical coverage. Here, we developed scalable manufacturing processes with a dense electrical connection scheme to achieve reconfigurable thin-film, multithousand-channel neurophysiological recording grids using platinum nanorods (PtNRGrids). With PtNRGrids, we have achieved a multithousand-channel array of small (30 µm) contacts with low impedance, providing high spatial and temporal resolution over a large cortical area. We demonstrated that PtNRGrids can resolve submillimeter functional organization of the barrel cortex in anesthetized rats that captured the tissue structure. In the clinical setting, PtNRGrids resolved fine, complex temporal dynamics from the cortical surface in an awake human patient performing grasping tasks. In addition, the PtNRGrids identified the spatial spread and dynamics of epileptic discharges in a patient undergoing epilepsy surgery at 1-mm spatial resolution, including activity induced by direct electrical stimulation. Collectively, these findings demonstrated the power of the PtNRGrids to transform clinical mapping and research with brain-machine interfaces.

Considerations and recent advances in nanoscale interfaces with neuronal and cardiac networks.

Nanoscale interfaces with biological tissue, principally made with nanowires (NWs), are envisioned as minimally destructive to the tissue and as scalable tools to directly transduce the electrochemical activity of a neuron at its finest resolution. This review lays the foundations for understanding the material and device considerations required to interrogate neuronal activity at the nanoscale. We first discuss the electrochemical nanoelectrode-neuron interfaces and then present new results concerning the electrochemical impedance and charge injection capacities of millimeter, micrometer, and nanometer scale wires with Pt, PEDOT:PSS, Si, Ti, ITO, IrO x , Ag, and AgCl materials. Using established circuit models for NW-neuron interfaces, we discuss the impact of having multiple NWs interfacing with a single neuron on the amplitude and temporal characteristics of the recorded potentials. We review state of the art advances in nanoelectrode-neuron interfaces, the standard control experiments to investigate their electrophysiological behavior, and present recent high fidelity recordings of intracellular potentials obtained with ultrasharp NWs developed in our laboratory that naturally permeate neuronal cell bodies. Recordings from arrays and individually addressable electrically shorted NWs are presented, and the long-term stability of intracellular recording is discussed and put in the context of established techniques. Finally, a perspective on future research directions and applications is presented.

An Analytical Model for Dual Gate Piezoelectrically Sensitive ZnO Thin Film Transistors.

We report the experimental realization of piezoelectric ZnO dual-gate thin film transistors (TFTs) as highly sensitive force sensors and discuss the physical origins of its electrically tunable piezoelectric response using a simple analytical model. A dual gate TFT is fabricated on a polyimide substrate using radio-frequency (RF) magnetron sputtering of piezoelectric ZnO thin film as a channel. The ZnO TFTs exhibited a field effect mobility of ~ 5 cm2/Vs, I max to I min ratio of 107, and a subthreshold slope of 700 mV/dec. Notably, the TFT exhibited static and transient current changes under external force stimuli, with varying amplitude and polarity for different gate bias regimes. To understand the current modulation of the dual-gate TFT with independently biased top and bottom gates, an analytical model is developed. The model includes accumulation channels at both surfaces and a bulk channel within the film and accounts for the force-induced piezoelectric charge density. The microscopic piezoelectric response that modulates the energy-band edges and correspondent current-voltage characteristics are accurately portrayed by our model. Finally, the field-tunable force response in single TFT is demonstrated as a function of independent bias for the top and bottom gates with a force response range from -0.29 nA/mN to 22.7 nA/mN. This work utilizes intuitive analytical models to shed light on the correlation between the material properties with the force response in piezoelectric TFTs.

Microscale dynamics of electrophysiological markers of epilepsy.

OBJECTIVE: Interictal discharges (IIDs) and high frequency oscillations (HFOs) are established neurophysiologic biomarkers of epilepsy, while microseizures are less well studied. We used custom poly(3,4-ethylenedioxythiophene) polystyrene sulfonate (PEDOT:PSS) microelectrodes to better understand these markers' microscale spatial dynamics. METHODS: Electrodes with spatial resolution down to 50 µm were used to record intraoperatively in 30 subjects. IIDs' degree of spread and spatiotemporal paths were generated by peak-tracking followed by clustering. Repeating HFO patterns were delineated by clustering similar time windows. Multi-unit activity (MUA) was analyzed in relation to IID and HFO timing. RESULTS: We detected IIDs encompassing the entire array in 93% of subjects, while localized IIDs, observed across < 50% of channels, were seen in 53%. IIDs traveled along specific paths. HFOs appeared in small, repeated spatiotemporal patterns. Finally, we identified microseizure events that spanned 50-100 µm. HFOs covaried with MUA, but not with IIDs. CONCLUSIONS: Overall, these data suggest that irritable cortex micro-domains may form part of an underlying pathologic architecture which could contribute to the seizure network. SIGNIFICANCE: These results, supporting the possibility that epileptogenic cortex comprises a mosaic of irritable domains, suggests that microscale approaches might be an important perspective in devising novel seizure control therapies.

Electrochemical safety limits for clinical stimulation investigated using depth and strip electrodes in the pig brain.

Objective. Diagnostic and therapeutic electrical stimulation are increasingly utilized with the rise of neuromodulation devices. However, systematic investigations that depict the practical clinical stimulation paradigms (bipolar, two-electrode configuration) to determine the safety limits are currently lacking. Further, safe charge densities that were classically determined from conical sharp electrodes are generalized for cylindrical (depth) and flat (surface grid) electrodes completely ignoring geometric factors that govern current spreading and trajectories in tissue.Approach. This work reports the first investigations comparing stimulation limits for clinically used electrodes in two mediums: in benchtop experiments in saline andin vivoin a single acute experiment in the pig brain. We experimentally determine the geometric factors, the water electrolysis windows, and the current safety limits from voltage transients, for the sEEG, depth and surface strip electrodes in both mediums. Using four-electrode and three-electrode configuration measurements and comprehensive circuit models that accurately depict our measurements, we delineate the various elements of the stimulation medium, including the tissue-electrode interface impedance spectra, the medium impedance and the bias-dependent change in the interface impedance as a function of stimulation parameters.Main results. The results of our systematics studies suggest that safe currents in clinical bipolar stimulation determinedin vivocan be as much as 24 times smaller than those determined from benchtop experiments (for depth electrodes at a 1 ms pulse duration). Our detailed circuit modeling attributes this drastic difference in safe limits to the greatly dissimilar electrode/tissue and electrode/saline impedances.Significance. We established the electrochemical safety limits for commonly used clinical electrodesin vivoand revealed by detailied electrochemical modeling how they differ from benchtop evaluation. We argue that electrochemical limits and currents are unique for each electrode, should be measuredin vivoaccording to the protocols established in this work, and should be accounted for while setting the stimulation parameters for clinical applications including for chronic applications.