Fiber-based Probes for Electrophysiology, Photometry, Optical and Electrical Stimulation, Drug Delivery, and Fast-Scan Cyclic Voltammetry In Vivo.

Recording and modulation of neuronal activity enables the study of brain function in health and disease. While translational neuroscience relies on electrical recording and modulation techniques, mechanistic studies in rodent models leverage genetic precision of optical methods, such as optogenetics and imaging of fluorescent indicators. In addition to electrical signal transduction, neurons produce and receive diverse chemical signals which motivate tools to probe and modulate neurochemistry. Although the past decade has delivered a wealth of technologies for electrophysiology, optogenetics, chemical sensing, and optical recording, combining these modalities within a single platform remains challenging. This work leverages materials selection and convergence fiber drawing to permit neural recording, electrical stimulation, optogenetics, fiber photometry, drug and gene delivery, and voltammetric recording of neurotransmitters within individual fibers. Composed of polymers and non-magnetic carbon-based conductors, these fibers are compatible with magnetic resonance imaging, enabling concurrent stimulation and whole-brain monitoring. Their utility is demonstrated in studies of the mesolimbic reward pathway by simultaneously interfacing with the ventral tegmental area and nucleus accumbens in mice and characterizing the neurophysiological effects of a stimulant drug. This study highlights the potential of these fibers to probe electrical, optical, and chemical signaling across multiple brain regions in both mechanistic and translational studies.

An Open-Source Mouse Chronic EEG Array System with High-Density MXene-Based Skull Surface Electrodes.

Electroencephalography (EEG) is an indispensable tool in epilepsy, sleep, and behavioral research. In rodents, EEG recordings are typically performed with metal electrodes that traverse the skull into the epidural space. In addition to requiring major surgery, intracranial EEG is difficult to perform for more than a few electrodes, is time-intensive, and confounds experiments studying traumatic brain injury. Here, we describe an open-source cost-effective refinement of this technique for chronic mouse EEG recording. Our alternative two-channel (EEG2) and sixteen-channel high-density EEG (HdEEG) arrays use electrodes made of the novel, flexible 2D nanomaterial titanium carbide (Ti3C2T x ) MXene. The MXene electrodes are placed on the surface of the intact skull and establish an electrical connection without conductive gel or paste. Fabrication and implantation times of MXene EEG electrodes are significantly shorter than the standard approach, and recorded resting baseline and epileptiform EEG waveforms are similar to those obtained with traditional epidural electrodes. Applying HdEEG to a mild traumatic brain injury (mTBI) model in mice of both sexes revealed that mTBI significantly increased spike-wave discharge (SWD) preictal network connectivity with frequencies of interest in the ß-spectral band (12-30 Hz). These findings indicate that the fabrication of MXene electrode arrays is a cost-effective, efficient technology for multichannel EEG recording in mice that obviates the need for skull-penetrating surgery. Moreover, increased preictal ß-frequency network connectivity may contribute to the development of early post-mTBI SWDs.

Evaluating and benchmarking the EEG signal quality of high-density, dry MXene-based electrode arrays against gelled Ag/AgCl electrodes.

Objective.To evaluate the signal quality of dry MXene-based electrode arrays (also termed 'MXtrodes') for electroencephalographic (EEG) recordings where gelled Ag/AgCl electrodes are a standard.Approach.We placed 4 × 4 MXtrode arrays and gelled Ag/AgCl electrodes on different scalp locations. The scalp was cleaned with alcohol and rewetted with saline before application. We recorded from both electrode types simultaneously while participants performed a vigilance task.Main results.The root mean squared amplitude of MXtrodes was slightly higher than that of Ag/AgCl electrodes (.24-1.94 uV). Most MXtrode pairs had slightly lower broadband spectral coherence (.05 to .1 dB) and Delta- and Theta-band timeseries correlation (.05 to .1 units) compared to the Ag/AgCl pair (p< .001). However, the magnitude of correlation and coherence was high across both electrode types. Beta-band timeseries correlation and spectral coherence were higher between neighboring MXtrodes in the array (.81 to .84 units) than between any other pair (.70 to .75 units). This result suggests the close spacing of the nearest MXtrodes (3 mm) more densely sampled high spatial-frequency topographies. Event-related potentials were more similar between MXtrodes (ρ⩾ .95) than equally spaced Ag/AgCl electrodes (ρ⩽ .77,p< .001). Dry MXtrode impedance (x̄= 5.15 KΩ cm2) was higher and more variable than gelled Ag/AgCl electrodes (x̄= 1.21 KΩ cm2,p< .001). EEG was also recorded on the scalp across diverse hair types.Significance.Dry MXene-based electrodes record EEG at a quality comparable to conventional gelled Ag/AgCl while requiring minimal scalp preparation and no gel. MXtrodes can record independent signals at a spatial density four times higher than conventional electrodes, including through hair, thus opening novel opportunities for research and clinical applications that could benefit from dry and higher-density configurations.

Effect of the deposition process on the stability of Ti3C2Tx MXene films for bioelectronics.

Ti3C2Tx MXene is emerging as the enabling material in a broad range of wearable and implantable medical technologies, thanks to its outstanding electrical, electrochemical, and optoelectronic properties, and its compatibility with high-throughput solution-based processing. While the prevalence of Ti3C2Tx MXene in biomedical research, and in particular bioelectronics, has steadily increased, the long-term stability and degradation of Ti3C2Tx MXene films have not yet been thoroughly investigated, limiting its use for chronic applications. Here, we investigate the stability of Ti3C2Tx films and electrodes under environmental conditions that are relevant to medical and bioelectronic technologies: storage in ambient atmosphere (shelf-life), submersion in saline (akin to the in vivo environment), and storage in a desiccator (low-humidity). Furthermore, to evaluate the effect of the MXene deposition method and thickness on the film stability in the different conditions, we compare thin (25 nm), and thick (1.0 µm) films and electrodes fabricated via spray-coating and blade-coating. Our findings indicate that film processing method and thickness play a significant role in determining the long-term performance of Ti3C2Tx films and electrodes, with highly aligned, thick films from blade coating remarkably retaining their conductivity, electrochemical impedance, and morphological integrity even after 30 days in saline. Our extensive spectroscopic analysis reveals that the degradation of Ti3C2Tx films in high-humidity environments is primarily driven by moisture intercalation, ingress, and film delamination, with evidence of only minimal to moderate oxidation.

Alerting attention is sufficient to induce a phase-dependent behavior that can be predicted by frontal EEG.

Recent studies suggest that attention is rhythmic. Whether that rhythmicity can be explained by the phase of ongoing neural oscillations, however, is still debated. We contemplate that a step toward untangling the relationship between attention and phase stems from employing simple behavioral tasks that isolate attention from other cognitive functions (perception/decision-making) and by localized monitoring of neural activity with high spatiotemporal resolution over the brain regions associated with the attentional network. In this study, we investigated whether the phase of electroencephalography (EEG) oscillations predicts alerting attention. We isolated the alerting mechanism of attention using the Psychomotor Vigilance Task, which does not involve a perceptual component, and collected high resolution EEG using novel high-density dry EEG arrays at the frontal region of the scalp. We identified that alerting attention alone is sufficient to induce a phase-dependent modulation of behavior at EEG frequencies of 3, 6, and 8 Hz throughout the frontal region, and we quantified the phase that predicts the high and low attention states in our cohort. Our findings disambiguate the relationship between EEG phase and alerting attention.

Wearable High-Density MXene-Bioelectronics for Neuromuscular Diagnostics, Rehabilitation, and Assistive Technologies.

High-density surface electromyography (HDsEMG) allows noninvasive muscle monitoring and disease diagnosis. Clinical translation of current HDsEMG technologies is hampered by cost, limited scalability, low usability, and minimal spatial coverage. Here, this study presents, validates, and demonstrates the broad clinical applicability of dry wearable MXene HDsEMG arrays (MXtrodes) fabricated from safe and scalable liquid-phase processing of Ti3 C2 Tx . The fabrication scheme allows easy customization of array geometry to match subject anatomy, while the gel-free and minimal skin preparation enhance usability and comfort. The low impedance and high conductivity of the MXtrode arrays allow detection of the activity of large muscle groups at higher quality and spatial resolution than state-of-the-art wireless electromyography  sensors, and in realistic clinical scenarios. To demonstrate the clinical applicability of MXtrodes in the context of neuromuscular diagnostics and rehabilitation, simultaneous HDsEMG and biomechanical mapping of muscle groups across the whole calf during various tasks, ranging from controlled contractions to walking is shown. Finally, the integration of HDsEMG acquired with MXtrodes with a machine learning pipeline and the accurate prediction of the phases of human gait are shown. The results underscore the advantages and translatability of MXene-based wearable bioelectronics for studying neuromuscular function and disease, as well as for precision rehabilitation.

Magnetoelectric Nanodiscs Enable Wireless Transgene-Free Neuromodulation.

Deep-brain stimulation (DBS) with implanted electrodes revolutionized treatment of movement disorders and empowered neuroscience studies. Identifying less invasive alternatives to DBS may further extend its clinical and research applications. Nanomaterial-mediated transduction of magnetic fields into electric potentials offers an alternative to invasive DBS. Here, we synthesize magnetoelectric nanodiscs (MENDs) with a core-double shell Fe3O4-CoFe2O4-BaTiO3 architecture with efficient magnetoelectric coupling. We find robust responses to magnetic field stimulation in neurons decorated with MENDs at a density of 1 µg/mm2 despite individual-particle potentials below the neuronal excitation threshold. We propose a model for repetitive subthreshold depolarization, which combined with cable theory, corroborates our findings in vitro and informs magnetoelectric stimulation in vivo. MENDs injected into the ventral tegmental area of genetically intact mice at concentrations of 1 mg/mL enable remote control of reward behavior, setting the stage for mechanistic optimization of magnetoelectric neuromodulation and inspiring its future applications in fundamental and translational neuroscience.

Multimodal, Multiscale Insights into Hippocampal Seizures Enabled by Transparent, Graphene-Based Microelectrode Arrays.

Hippocampal seizures are a defining feature of mesial temporal lobe epilepsy (MTLE). Area CA1 of the hippocampus is commonly implicated in the generation of seizures, which may occur because of the activity of endogenous cell populations or of inputs from other regions within the hippocampal formation. Simultaneously observing activity at the cellular and network scales in vivo remains challenging. Here, we present a novel technology for simultaneous electrophysiology and multicellular calcium imaging of CA1 pyramidal cells (PCs) in mice enabled by a transparent graphene-based microelectrode array (Gr MEA). We examine PC firing at seizure onset, oscillatory coupling, and the dynamics of the seizure traveling wave as seizures evolve. Finally, we couple features derived from both modalities to predict the speed of the traveling wave using bootstrap aggregated regression trees. Analysis of the most important features in the regression trees suggests a transition among states in the evolution of hippocampal seizures.

Emerging approaches for sensing and modulating neural activity enabled by nanocarbons and carbides.

Devices that can record or modulate neural activity are essential tools in clinical diagnostics and monitoring, basic research, and consumer electronics. Realizing stable functional interfaces between manmade electronics and biological tissues is a longstanding challenge that requires device and material innovations to meet stringent safety and longevity requirements and to improve functionality. Compared to conventional materials, nanocarbons and carbides offer a number of specific advantages for neuroelectronics that can enable advances in functionality and performance. Here, we review the latest emerging trends in neuroelectronic interfaces based on nanocarbons and carbides, with a specific emphasis on technologies developed for use in vivo. We highlight specific applications where the ability to tune fundamental material properties at the nanoscale enables interfaces that can safely and precisely interact with neural circuits at unprecedented spatial and temporal scales, ranging from single synapses to the whole human body.

MXene-infused bioelectronic interfaces for multiscale electrophysiology and stimulation.

Soft bioelectronic interfaces for mapping and modulating excitable networks at high resolution and at large scale can enable paradigm-shifting diagnostics, monitoring, and treatment strategies. Yet, current technologies largely rely on materials and fabrication schemes that are expensive, do not scale, and critically limit the maximum attainable resolution and coverage. Solution processing is a cost-effective manufacturing alternative, but biocompatible conductive inks matching the performance of conventional metals are lacking. Here, we introduce MXtrodes, a class of soft, high-resolution, large-scale bioelectronic interfaces enabled by Ti3C2 MXene (a two-dimensional transition metal carbide nanomaterial) and scalable solution processing. We show that the electrochemical properties of MXtrodes exceed those of conventional materials and do not require conductive gels when used in epidermal electronics. Furthermore, we validate MXtrodes in applications ranging from mapping large-scale neuromuscular networks in humans to cortical neural recording and microstimulation in swine and rodent models. Last, we demonstrate that MXtrodes are compatible with standard clinical neuroimaging modalities.

Multimodal in vivo recording using transparent graphene microelectrodes illuminates spatiotemporal seizure dynamics at the microscale.

Neurological disorders such as epilepsy arise from disrupted brain networks. Our capacity to treat these disorders is limited by our inability to map these networks at sufficient temporal and spatial scales to target interventions. Current best techniques either sample broad areas at low temporal resolution (e.g. calcium imaging) or record from discrete regions at high temporal resolution (e.g. electrophysiology). This limitation hampers our ability to understand and intervene in aberrations of network dynamics. Here we present a technique to map the onset and spatiotemporal spread of acute epileptic seizures in vivo by simultaneously recording high bandwidth microelectrocorticography and calcium fluorescence using transparent graphene microelectrode arrays. We integrate dynamic data features from both modalities using non-negative matrix factorization to identify sequential spatiotemporal patterns of seizure onset and evolution, revealing how the temporal progression of ictal electrophysiology is linked to the spatial evolution of the recruited seizure core. This integrated analysis of multimodal data reveals otherwise hidden state transitions in the spatial and temporal progression of acute seizures. The techniques demonstrated here may enable future targeted therapeutic interventions and novel spatially embedded models of local circuit dynamics during seizure onset and evolution.

Gels, jets, mosquitoes, and magnets: a review of implantation strategies for soft neural probes.

Implantable neuroelectronic interfaces have enabled breakthrough advances in the clinical diagnosis and treatment of neurological disorders, as well as in fundamental studies of brain function, behavior, and disease. Intracranial electroencephalography (EEG) mapping with stereo-EEG (sEEG) depth electrodes is routinely adopted for precise epilepsy diagnostics and surgical treatment, while deep brain stimulation has become the standard of care for managing movement disorders. Intracortical microelectrode arrays for high-fidelity recordings of neural spiking activity have led to impressive demonstrations of the power of brain-machine interfaces for motor and sensory functional recovery. Yet, despite the rapid pace of technology development, the issue of establishing a safe, long-term, stable, and functional interface between neuroelectronic devices and the host brain tissue still remains largely unresolved. A body of work spanning at least the last 15 years suggests that safe, chronic integration between invasive electrodes and the brain requires a close match between the mechanical properties of man-made components and the neural tissue. In other words, the next generation of invasive electrodes should be soft and compliant, without sacrificing biological and chemical stability. Soft neuroelectronic interfaces, however, pose a new and significant surgical challenge: bending and buckling during implantation that can preclude accurate and safe device placement. In this topical review, we describe the next generation of soft electrodes and the surgical implantation methods for safe and precise insertion into brain structures. We provide an overview of the most recent innovations in the field of insertion strategies for flexible neural electrodes such as dissolvable or biodegradable carriers, microactuators, biologically-inspired support structures, and electromagnetic drives. In our analysis, we also highlight approaches developed in different fields, such as robotic surgery, which could be potentially adapted and translated to the insertion of flexible neural probes.

Fabrication of Ti3C2 MXene Microelectrode Arrays for In Vivo Neural Recording.

Implantable microelectrode technologies have been widely used to elucidate neural dynamics at the microscale to gain a deeper understanding of the neural underpinnings of brain disease and injury. As electrodes are miniaturized to the scale of individual cells, a corresponding rise in the interface impedance limits the quality of recorded signals. Additionally, conventional electrode materials are stiff, resulting in a significant mechanical mismatch between the electrode and the surrounding brain tissue, which elicits an inflammatory response that eventually leads to a degradation of the device performance. To address these challenges, we have developed a process to fabricate flexible microelectrodes based on Ti3C2 MXene, a recently discovered nanomaterial that possesses remarkably high volumetric capacitance, electrical conductivity, surface functionality, and processability in aqueous dispersions. Flexible arrays of Ti3C2 MXene microelectrodes have remarkably low impedance due to the high conductivity and high specific surface area of the Ti3C2 MXene films, and they have proven to be exquisitely sensitive for recording neuronal activity. In this protocol, we describe a novel method for micropatterning Ti3C2 MXene into microelectrode arrays on flexible polymeric substrates and outline their use for in vivo micro-electrocorticography recording. This method can easily be extended to create MXene electrode arrays of arbitrary size or geometry for a range of other applications in bioelectronics and it can also be adapted for use with other conductive inks besides Ti3C2 MXene. This protocol enables simple and scalable fabrication of microelectrodes from solution-based conductive inks, and specifically allows harnessing the unique properties of hydrophilic Ti3C2 MXene to overcome many of the barriers that have long hindered the widespread adoption of carbon-based nanomaterials for high-fidelity neural microelectrodes.

A gel-free Ti3C2Tx-based electrode array for high-density, high-resolution surface electromyography.

Wearable sensors for surface electromyography (EMG) are composed of single- to few-channel large-area contacts, which exhibit high interfacial impedance and require conductive gels or adhesives to record high-fidelity signals. These devices are also limited in their ability to record activation across large muscle groups due to poor spatial coverage. To address these challenges, we have developed a novel high-density EMG array based on titanium carbide (Ti3C2Tx) MXene encapsulated in parylene-C. Ti3C2Tx is a two-dimensional nanomaterial with excellent electrical, electrochemical, and mechanical properties, which forms colloidally stable aqueous dispersions, enabling safe, scalable solutions-processing. Leveraging the excellent combination of metallic conductivity, high pseudocapacitance, and ease of processability of Ti3C2Tx MXene, we demonstrate the fabrication of gel-free, high-density EMG arrays which are ~8 µm thick, feature 16 recording channels, and are highly skin-conformable. The impedance of Ti3C2Tx electrodes in contact with human skin is 100-1000x lower than the impedance of commercially-available electrodes which require conductive gels to be effective. Furthermore, our arrays can record high-fidelity, low-noise EMG, and can resolve muscle activation with improved spatiotemporal resolution and sensitivity compared to conventional gelled electrodes. Overall, our results establish Ti3C2Tx-based bioelectronic interfaces as a powerful platform technology for high-resolution, non-invasive wearable sensing technologies.

Biomimetic extracellular matrix coatings improve the chronic biocompatibility of microfabricated subdural microelectrode arrays.

Intracranial electrodes are a vital component of implantable neurodevices, both for acute diagnostics and chronic treatment with open and closed-loop neuromodulation. Their performance is hampered by acute implantation trauma and chronic inflammation in response to implanted materials and mechanical mismatch between stiff synthetic electrodes and pulsating, natural soft host neural tissue. Flexible electronics based on thin polymer films patterned with microscale conductive features can help alleviate the mechanically induced trauma; however, this strategy alone does not mitigate inflammation at the device-tissue interface. In this study, we propose a biomimetic approach that integrates microscale extracellular matrix (ECM) coatings on microfabricated flexible subdural microelectrodes. Taking advantage of a high-throughput process employing micro-transfer molding and excimer laser micromachining, we fabricate multi-channel subdural microelectrodes primarily composed of ECM protein material and demonstrate that the electrochemical and mechanical properties match those of standard, uncoated controls. In vivo ECoG recordings in rodent brain confirm that the ECM microelectrode coatings and the protein interface do not alter signal fidelity. Astrogliotic, foreign body reaction to ECM coated devices is reduced, compared to uncoated controls, at 7 and 30 days, after subdural implantation in rat somatosensory cortex. We propose microfabricated, flexible, biomimetic electrodes as a new strategy to reduce inflammation at the device-tissue interface and improve the long-term stability of implantable subdural electrodes.

Two-Dimensional Ti3C2 MXene for High-Resolution Neural Interfaces.

High-resolution neural interfaces are essential tools for studying and modulating neural circuits underlying brain function and disease. Because electrodes are miniaturized to achieve higher spatial resolution and channel count, maintaining low impedance and high signal quality becomes a significant challenge. Nanostructured materials can address this challenge because they combine high electrical conductivity with mechanical flexibility and can interact with biological systems on a molecular scale. Unfortunately, fabricating high-resolution neural interfaces from nanostructured materials is typically expensive and time-consuming and does not scale, which precludes translation beyond the benchtop. Two-dimensional (2D) Ti3C2 MXene possesses a combination of remarkably high volumetric capacitance, electrical conductivity, surface functionality, and processability in aqueous dispersions distinct among carbon-based nanomaterials. Here, we present a high-throughput microfabrication process for constructing Ti3C2 neuroelectronic devices and demonstrate their superior impedance and in vivo neural recording performance in comparison with standard metal microelectrodes. Specifically, when compared to gold microelectrodes of the same size, Ti3C2 electrodes exhibit a 4-fold reduction in interface impedance. Furthermore, intraoperative in vivo recordings from the brains of anesthetized rats at multiple spatial and temporal scales demonstrate that Ti3C2 electrodes exhibit lower baseline noise, higher signal-to-noise ratio, and reduced susceptibility to 60 Hz interference than gold electrodes. Finally, in neuronal biocompatibility studies, neurons cultured on Ti3C2 are as viable as those in control cultures, and they can adhere, grow axonal processes, and form functional networks. Overall, our results indicate that Ti3C2 MXene microelectrodes have the potential to become a powerful platform technology for high-resolution biological interfaces.

Modulating dopamine release by optogenetics in transgenic mice reveals terminal dopaminergic dynamics.

Dopamine (DA) release and uptake dynamics in the nucleus accumbens (NAc) have important implications for neurological diseases and mammalian animal behaviors. We demonstrate here the use of cell-type-specific optogenetic targeting in conjunction with fast-scan cyclic voltammetry applied to brain slices prepared from specifically tailored transgenic mice, which conditionally express channelrhodopsin-2 (ChR2) through dopamine transporter (DAT)-Cre. Terminal dopaminergic dynamics and the direct manipulation of induced DA release level by controlling light intensity, pulse width, and the shape of stimulation waveforms were studied. Effective cell terminal-targeting optogenetic induction of DA release at physiological levels in NAc is demonstrated and discussed. It was found that delivering more light energy by increasing stimulation intensity and length is not the only way to control DA release; the temporal shape of the stimulus waveform at light onset is also critically related to induced DA concentrations. In addition, DA uptake dynamics as well as the recovery of the presynaptic releasable DA pool are studied and modeled. More broadly, our experimental findings provide important further evidence for effectively applying optogenetics to induce neurotransmitter release in the behaviorally relevant region of the brain in a highly cell-type selective context.

Thromboelastography defines late hypercoagulability after TBI: a pilot study.

INTRODUCTION: Traumatic brain injury (TBI) is associated with a hypercoagulable state, the mechanism and duration of which remain unclear. We sought to determine whether thromboelastography (TEG) analysis could identify the hypercoagulable state after TBI, as defined by elevations in maximal amplitude (MA), thrombus generation (TG), G value (G), and alpha angle (αA). METHODS: Patients with moderate-severe TBI, defined primarily as a GCS <12, admitted between 1/2012 and 8/2013 were eligible for enrolment in this prospective cohort study. TEG profiles were obtained between 0-24 h (T1), 24-48 h (T2), 48-72 h (T3), 72-96 h (T4), and 96-120 h (T5) after admission. Early TEG was defined as 0-48 h, and late TEG was defined as >48 h. RESULTS: Twenty five patients (80 % men) and 7 age- and sex-matched control subjects were studied. Median age was 38 years (range 18-85). Early MA was [63.6 mm (60.5, 67.4)] versus late MA [69.9 mm (65.2,73.9); p = 0.02], early TG was [763.3 mm/min (712.8, 816.2)] versus late TG [835.9 mm/min (791.2,888.3); p = 0.02], and early G was [8.8 d/cm(2) (7.7,10.4)] versus late G [11.6 d/cm(2) (9.4,14.1); p = 0.02]. Study patients had higher MA (p = 0.02), TG (p = 0.03), and G (p = 0.02) values at T5 compared to controls. There was a linear increase per day of MA by 2.6 mm (p = 0.001), TG 31.9 mm/min (p ≤ 0.001), and G value by 1.3 d/cm(2) (p ≤ 0.001) when clustered by pairs in regression analysis. Lower MA values trended toward home discharge (p = 0.08). CONCLUSION: The data suggest a progressive and delayed hypercoagulable state observed days after initial TBI. The hypercoagulable state may reflect excess platelet activity.