Ultraflexible and Stretchable Intrafascicular Peripheral Nerve Recording Device with Axon-Dimension, Cuff-Less Microneedle Electrode Array.

Peripheral nerve mapping tools with higher spatial resolution are needed to advance systems neuroscience, and potentially provide a closed-loop biomarker in neuromodulation applications. Two critical challenges of microscale neural interfaces are 1) how to apply them to small peripheral nerves, and 2) how to minimize chronic reactivity. A flexible microneedle nerve array (MINA) is developed, which is the first high-density penetrating electrode array made with axon-sized silicon microneedles embedded in low-modulus thin silicone. The design, fabrication, acute recording, and chronic reactivity to an implanted MINA, are presented. Distinctive units are identified in the rat peroneal nerve. The authors also demonstrate a long-term, cuff-free, and suture-free fixation manner using rose bengal as a light-activated adhesive for two time-points. The tissue response is investigated at 1-week and 6-week time-points, including two sham groups and two MINA-implanted groups. These conditions are quantified in the left vagus nerve of rats using histomorphometry. Micro computed tomography (micro-CT) is added to visualize and quantify tissue encapsulation around the implant. MINA demonstrates a reduction in encapsulation thickness over previously quantified interfascicular methods. Future challenges include techniques for precise insertion of the microneedle electrodes and demonstrating long-term recording.

A Materials Roadmap to Functional Neural Interface Design.

Advancement in neurotechnologies for electrophysiology, neurochemical sensing, neuromodulation, and optogenetics are revolutionizing scientific understanding of the brain while enabling treatments, cures, and preventative measures for a variety of neurological disorders. The grand challenge in neural interface engineering is to seamlessly integrate the interface between neurobiology and engineered technology, to record from and modulate neurons over chronic timescales. However, the biological inflammatory response to implants, neural degeneration, and long-term material stability diminish the quality of interface overtime. Recent advances in functional materials have been aimed at engineering solutions for chronic neural interfaces. Yet, the development and deployment of neural interfaces designed from novel materials have introduced new challenges that have largely avoided being addressed. Many engineering efforts that solely focus on optimizing individual probe design parameters, such as softness or flexibility, downplay critical multi-dimensional interactions between different physical properties of the device that contribute to overall performance and biocompatibility. Moreover, the use of these new materials present substantial new difficulties that must be addressed before regulatory approval for use in human patients will be achievable. In this review, the interdependence of different electrode components are highlighted to demonstrate the current materials-based challenges facing the field of neural interface engineering.

Sensing local field potentials with a directional and scalable depth electrode array.

Objective. A variety of electrophysiology tools are available to the neurosurgeon for diagnosis, functional therapy, and neural prosthetics. However, no tool can currently address these three critical needs: (a) access to all cortical regions in a minimally invasive manner; (b) recordings with microscale, mesoscale, and macroscale resolutions simultaneously; and (c) access to spatially distant multiple brain regions that constitute distributed cognitive networks.Approach.We modeled, designed, and demonstrated a novel device for recording local field potentials (LFPs) with the form factor of a stereo-electroencephalographic electrode and combined with radially distributed microelectrodes.Main results. Electro-quasistatic models demonstrate that the lead body amplifies and shields LFP sources based on direction, enablingdirectional sensitivity andscalability, referred to as thedirectional andscalable (DISC) array.In vivo,DISC demonstrated significantly improved signal-to-noise ratio, directional sensitivity, and decoding accuracy from rat barrel cortex recordings during whisker stimulation. Critical for future translation, DISC demonstrated a higher signal to noise ratio (SNR) than virtual ring electrodes and a noise floor approaching that of large ring electrodes in an unshielded environment after common average referencing. DISC also revealed independent, stereoscopic current source density measures whose direction was verified after histology.Significance. Directional sensitivity of LFPs may significantly improve brain-computer interfaces and many diagnostic procedures, including epilepsy foci detection and deep brain targeting.

Decoding articulatory and phonetic components of naturalistic continuous speech from the distributed language network.

Objective.The speech production network relies on a widely distributed brain network. However, research and development of speech brain-computer interfaces (speech-BCIs) has typically focused on decoding speech only from superficial subregions readily accessible by subdural grid arrays-typically placed over the sensorimotor cortex. Alternatively, the technique of stereo-electroencephalography (sEEG) enables access to distributed brain regions using multiple depth electrodes with lower surgical risks, especially in patients with brain injuries resulting in aphasia and other speech disorders.Approach.To investigate the decoding potential of widespread electrode coverage in multiple cortical sites, we used a naturalistic continuous speech production task. We obtained neural recordings using sEEG from eight participants while they read aloud sentences. We trained linear classifiers to decode distinct speech components (articulatory components and phonemes) solely based on broadband gamma activity and evaluated the decoding performance using nested five-fold cross-validation.Main Results.We achieved an average classification accuracy of 18.7% across 9 places of articulation (e.g. bilabials, palatals), 26.5% across 5 manner of articulation (MOA) labels (e.g. affricates, fricatives), and 4.81% across 38 phonemes. The highest classification accuracies achieved with a single large dataset were 26.3% for place of articulation, 35.7% for MOA, and 9.88% for phonemes. Electrodes that contributed high decoding power were distributed across multiple sulcal and gyral sites in both dominant and non-dominant hemispheres, including ventral sensorimotor, inferior frontal, superior temporal, and fusiform cortices. Rather than finding a distinct cortical locus for each speech component, we observed neural correlates of both articulatory and phonetic components in multiple hubs of a widespread language production network.Significance.These results reveal the distributed cortical representations whose activity can enable decoding speech components during continuous speech through the use of this minimally invasive recording method, elucidating language neurobiology and neural targets for future speech-BCIs.

HectoSTAR µLED Optoelectrodes for Large-Scale, High-Precision In Vivo Opto-Electrophysiology.

Dynamic interactions within and across brain areas underlie behavioral and cognitive functions. To understand the basis of these processes, the activities of distributed local circuits inside the brain of a behaving animal must be synchronously recorded while the inputs to these circuits are precisely manipulated. Even though recent technological advances have enabled such large-scale recording capabilities, the development of the high-spatiotemporal-resolution and large-scale modulation techniques to accompany those recordings has lagged. A novel neural probe is presented in this work that enables simultaneous electrical monitoring and optogenetic manipulation of deep neuronal circuits at large scales with a high spatiotemporal resolution. The "hectoSTAR" micro-light-emitting-diode (µLED) optoelectrode features 256 recording electrodes and 128 stimulation µLEDs monolithically integrated on the surface of its four 30-µm thick silicon micro-needle shanks, covering a large volume with 1.3-mm × 0.9-mm cross-sectional area located as deep as 6 mm inside the brain. The use of this device in behaving mice for dissecting long-distance network interactions across cortical layers and hippocampal regions is demonstrated. The recording-and-stimulation capabilities hectoSTAR µLED optoelectrodes enables will open up new possibilities for the cellular and circuit-based investigation of brain functions in behaving animals.

Novel multi-sided, microelectrode arrays for implantable neural applications.

A new parylene-based microfabrication process is presented for neural recording and drug delivery applications. We introduce a large design space for electrode placement and structural flexibility with a six mask process. By using chemical mechanical polishing, electrode sites may be created top-side, back-side, or on the edge of the device having three exposed sides. Added surface area was achieved on the exposed edge through electroplating. Poly(3,4-ethylenedioxythiophene) (PEDOT) modified edge electrodes having an 85-µm(2) footprint resulted in an impedance of 200 kΩ at 1 kHz. Edge electrodes were able to successfully record single unit activity in acute animal studies. A finite element model of planar and edge electrodes relative to neuron position reveals that edge electrodes should be beneficial for increasing the volume of tissue being sampled in recording applications.

High-density neural recordings from feline sacral dorsal root ganglia with thin-film array.

Objective. Dorsal root ganglia (DRG) are promising sites for recording sensory activity. Current technologies for DRG recording are stiff and typically do not have sufficient site density for high-fidelity neural data techniques.Approach. In acute experiments, we demonstrate single-unit neural recordings in sacral DRG of anesthetized felines using a 4.5µm thick, high-density flexible polyimide microelectrode array with 60 sites and 30-40µm site spacing. We delivered arrays into DRG with ultrananocrystalline diamond shuttles designed for high stiffness affording a smaller footprint. We recorded neural activity during sensory activation, including cutaneous brushing and bladder filling, as well as during electrical stimulation of the pudendal nerve and anal sphincter. We used specialized neural signal analysis software to sort densely packed neural signals.Main results. We successfully delivered arrays in five of six experiments and recorded single-unit sensory activity in four experiments. The median neural signal amplitude was 55µV peak-to-peak and the maximum unique units recorded at one array position was 260, with 157 driven by sensory or electrical stimulation. In one experiment, we used the neural analysis software to track eight sorted single units as the array was retracted ∼500µm.Significance. This study is the first demonstration of ultrathin, flexible, high-density electronics delivered into DRG, with capabilities for recording and tracking sensory information that are a significant improvement over conventional DRG interfaces.

Sharpened and Mechanically Durable Carbon Fiber Electrode Arrays for Neural Recording.

Bioelectric medicine treatments target disorders of the nervous system unresponsive to pharmacological methods. While current stimulation paradigms effectively treat many disorders, the underlying mechanisms are relatively unknown, and current neuroscience recording electrodes are often limited in their specificity to gross averages across many neurons or axons. Here, we develop a novel, durable carbon fiber electrode array adaptable to many neural structures for precise neural recording. Carbon fibers ( [Formula: see text] diameter) were sharpened using a reproducible blowtorchmethod that uses the reflection of fibers against the surface of a water bath. The arrays were developed by partially embedding carbon fibers in medical-grade silicone to improve durability. We recorded acute spontaneous electrophysiology from the rat cervical vagus nerve (CVN), feline dorsal root ganglia (DRG), and rat brain. Blowtorching resulted in fibers of 72.3 ± 33.5-degree tip angle with [Formula: see text] exposed carbon. Observable neural clusters were recorded using sharpened carbon fiber electrodes fromrat CVN ( [Formula: see text]), feline DRG ( [Formula: see text]), and rat brain ( [Formula: see text]). Recordings from the feline DRG included physiologically relevant signals from increased bladder pressure and cutaneous brushing. These results suggest that this carbon fiber array is a uniquely durable and adaptable neural recordingdevice. In the future, this device may be useful as a bioelectric medicine tool for diagnosis and closed-loop neural control of therapeutic treatments and monitoring systems.

Artifact-free and high-temporal-resolution in vivo opto-electrophysiology with microLED optoelectrodes.

The combination of in vivo extracellular recording and genetic-engineering-assisted optical stimulation is a powerful tool for the study of neuronal circuits. Precise analysis of complex neural circuits requires high-density integration of multiple cellular-size light sources and recording electrodes. However, high-density integration inevitably introduces stimulation artifact. We present minimal-stimulation-artifact (miniSTAR) µLED optoelectrodes that enable effective elimination of stimulation artifact. A multi-metal-layer structure with a shielding layer effectively suppresses capacitive coupling of stimulation signals. A heavily boron-doped silicon substrate silences the photovoltaic effect induced from LED illumination. With transient stimulation pulse shaping, we reduced stimulation artifact on miniSTAR µLED optoelectrodes to below 50 µVpp, much smaller than a typical spike detection threshold, at optical stimulation of >50 mW mm-2 irradiance. We demonstrated high-temporal resolution (<1 ms) opto-electrophysiology without any artifact-induced signal quality degradation during in vivo experiments. MiniSTAR µLED optoelectrodes will facilitate functional mapping of local circuits and discoveries in the brain.

Novel diamond shuttle to deliver flexible neural probe with reduced tissue compression.

The ability to deliver flexible biosensors through the toughest membranes of the central and peripheral nervous system is an important challenge in neuroscience and neural engineering. Bioelectronic devices implanted through dura mater and thick epineurium would ideally create minimal compression and acute damage as they reach the neurons of interest. We demonstrate that a three-dimensional diamond shuttle can be easily made with a vertical support to deliver ultra-compliant polymer microelectrodes (4.5-µm thick) through dura mater and thick epineurium. The diamond shuttle has 54% less cross-sectional area than an equivalently stiff silicon shuttle, which we simulated will result in a 37% reduction in blood vessel damage. We also discovered that higher frequency oscillation of the shuttle (200 Hz) significantly reduced tissue compression regardless of the insertion speed, while slow speeds also independently reduced tissue compression. Insertion and recording performance are demonstrated in rat and feline models, but the large design space of these tools are suitable for research in a variety of animal models and nervous system targets.

Multi-channel intraneural vagus nerve recordings with a novel high-density carbon fiber microelectrode array.

Autonomic nerves convey essential neural signals that regulate vital body functions. Recording clearly distinctive physiological neural signals from autonomic nerves will help develop new treatments for restoring regulatory functions. However, this is very challenging due to the small nature of autonomic nerves and the low-amplitude signals from their small axons. We developed a multi-channel, high-density, intraneural carbon fiber microelectrode array (CFMA) with ultra-small electrodes (8-9 µm in diameter, 150-250 µm in length) for recording physiological action potentials from small autonomic nerves. In this study, we inserted CFMA with up to 16 recording carbon fibers in the cervical vagus nerve of 22 isoflurane-anesthetized rats. We recorded action potentials with peak-to-peak amplitudes of 15.1-91.7 µV and signal-to-noise ratios of 2.0-8.3 on multiple carbon fibers per experiment, determined conduction velocities of some vagal signals in the afferent (0.7-4.4 m/s) and efferent (0.7-8.8 m/s) directions, and monitored firing rate changes in breathing and blood glucose modulated conditions. Overall, these experiments demonstrated that CFMA is a novel interface for in-vivo intraneural action potential recordings. This work is considerable progress towards the comprehensive understanding of physiological neural signaling in vital regulatory functions controlled by autonomic nerves.

Ultracompliant Carbon Nanotube Direct Bladder Device.

The bladder, stomach, intestines, heart, and lungs all move dynamically to achieve their purpose. A long-term implantable device that can attach onto an organ, sense its movement, and deliver current to modify the organ function would be useful in many therapeutic applications. The bladder, for example, can suffer from incomplete contractions that result in urinary retention with patients requiring catheterization. Those affected may benefit from a combination of a strain sensor and electrical stimulator to better control bladder emptying. The materials and design of such a device made from thin layer carbon nanotube (CNT) and Ecoflex 00-50 are described and demonstrate its function with in vivo feline bladders. During bench-top characterization, the resistive and capacitive sensors exhibit stability throughout 5000 stretching cycles under physiology conditions. In vivo measurements with piezoresistive devices show a high correlation between sensor resistance and volume. Stimulation driven from platinum-silicone composite electrodes successfully induce bladder contraction. A method for reliable connection and packaging of medical grade wire to the CNT device is also presented. This work is an important step toward the translation of low-durometer elastomers, stretchable CNT percolation, and platinum-silicone composite, which are ideal for large-strain bioelectric applications to sense or modulate dynamic organ states.

A High-Resolution Opto-Electrophysiology System With a Miniature Integrated Headstage.

This work presents a fully integrated neural interface system in a small form factor (1.9 g), consisting of a µLED silicon optoelectrode (12 µLEDs and 32 recording sites in a 4-shank configuration), an Intan 32-channel recording chip, and a custom optical stimulation chip for controlling 12 µLEDs. High-resolution optical stimulation with approximately 68.5 nW radiant flux resolution is achieved by a custom LED driver ASIC, which enables individual control of up to 48 channels with a current precision of 1 µA, a maximum current of 1.024 mA, and an update rate of >10 kHz. Recording is performed by an off-the-shelf 32-channel digitizing front-end ASIC from Intan. Two compact custom interface printed circuit boards were designed to link the headstage with a PC. The prototype system demonstrates precise current generation, sufficient optical radiant flux generation , and fast turn-on of µLEDs . Single animal in vivo experiments validated the headstage's capability to precisely modulate single neuronal activity and independently modulate activities of separate neuronal populations near neighboring optoelectrode shanks.

Neural probe design for reduced tissue encapsulation in CNS.

This study investigated relationships between a microscale neural probe's size and shape and its chronic reactive tissue response. Parylene-based probes were microfabricated with a thick shank (48 microm by 68 microm) and an integrated thin lateral platform (5 microm by 100 microm, either solid or one of three lattice sizes). Devices were implanted in rat cerebral cortex for 4 weeks before immunostaining for neurons, astrocytes, microglia, fibronectin, laminin, and neurofilament. While nonneuronal density (NND) generally increased and neuronal density decreased within 75 microm of a probe interface compared to unimplanted control regions, there were significant differential tissue responses within 25 microm of the platform's lateral edge compared to the shank. The NND in this region of the lateral edge was less than one-third of the corresponding region of the shank (129% and 425% increase, respectively). Moreover, neuronal density around the platform lateral edge was about one-third higher than at the shank (0.70 and 0.52 relative to control, respectively). Also, microglia reactivity and extracellular protein deposition was reduced at the lateral edge. There were no significant differences among platform designs. These results suggest that neural probe geometry is an important parameter for reducing chronic tissue encapsulation.

State-of-the-art MEMS and microsystem tools for brain research.

Mapping brain activity has received growing worldwide interest because it is expected to improve disease treatment and allow for the development of important neuromorphic computational methods. MEMS and microsystems are expected to continue to offer new and exciting solutions to meet the need for high-density, high-fidelity neural interfaces. Herein, the state-of-the-art in recording and stimulation tools for brain research is reviewed, and some of the most significant technology trends shaping the field of neurotechnology are discussed.

Investigation of the photoelectrochemical effect in optoelectrodes and potential uses for implantable electrode characterization.

The combination of optical stimulation and electrical recording is commonly employed in neuroscience research. Researchers using optogenetics are familiar with the photo-induced "artifacts" that arise from illumination of an electrode array. We sought to characterize this photoelectrochemical (PEC) effect to understand the underlying mechanism seen on NeuroNexus optoelectrodes. In doing so, we discovered that the phenomenon is inversely proportional to electrode site area in the same manner as electrical impedance measurements. We have applied the PEC effect as a method of electrode evaluation and show that a PEC measurement system can be both faster and more effective than impedance at sensing defects in high-throughput biomedical device testing.

The insulation performance of reactive parylene films in implantable electronic devices.

Parylene-C (poly-chloro-p-xylylene) is an appropriate material for use in an implantable, microfabricated device. It is hydrophobic, conformally deposited, has a low dielectric constant, and superb biocompatibility. Yet for many bioelectrical applications, its poor wet adhesion may be an impassable shortcoming. This research contrasts parylene-C and poly(p-xylylene) functionalized with reactive group X (PPX-X) layers using long-term electrical soak and adhesion tests. The reactive parylene was made of complementary derivatives having aldehyde and aminomethyl side groups (PPX-CHO and PPX-CH2NH2 respectively). These functional groups have previously been shown to covalently react together after heating. Electrical testing was conducted in saline at 37 degrees C on interdigitated electrodes with either parylene-C or reactive parylene as the metal layer interface. Results showed that reactive parylene devices maintained the highest impedance. Heat-treated PPX-X device impedance was 800% greater at 10kHz and 70% greater at 1Hz relative to heated parylene-C controls after 60 days. Heat treatment proved to be critical for maintaining high impedance of both parylene-C and the reactive parylene. Adhesion measurements showed improved wet metal adhesion for PPX-X, which corresponds well with its excellent high frequency performance.

Fabrication of polymer neural probes with sub-cellular features for reduced tissue encapsulation.

Intracortical microelectrodes currently have great potential as a neural prosthesis in patients with neurodegenerative disease or spinal cord injury. In an effort to improve the consistency of neural probe performance, many modifications to probe design are focused on reducing the tissue encapsulation. Since researchers have shown that small polymer fibers (less than 7-microm diameter) induce a small to non-existent encapsulation layer in the rat subcutis, we have proposed a neural probe design with similarly small diameter structures. This paper discusses the fabrication and design parameters of a microscale neural probe with a sub-cellular lattice structure. We developed a microfabrication process using SU-8 and parylene-C to create the relatively thick probe shank and thin lateral arms. The stiff penetrating shank (70-microm by 42-microm) had an SU-8 core that allowed control over stiffness and simplified the process. Parylene-only structures lateral to the shank could be defined with a 4-microm feature-size to meet our sub-cellular criterion. We fabricated four varying geometries for implantation into the neocortex of seven Sprague-Dawley rats. Our in vivo testing verifies that despite the flexible substrate and small dimensions (4-microm x 5-microm), these devices are mechanically robust and practical as neural probes. These devices provide an important tool for neural engineers to investigate the tissue response around sub-cellular structures and potentially improve device efficacy.