Flexible Microelectrode Array Research Articles

Flexible high-density microelectrode arrays for closed-loop brain-machine interfaces: a review.

Flexible high-density microelectrode arrays (HDMEAs) are emerging as a key component in closed-loop brain-machine interfaces (BMIs), providing high-resolution functionality for recording, stimulation, or both. The flexibility of these arrays provides advantages over rigid ones, such as reduced mismatch between interface and tissue, resilience to micromotion, and sustained long-term performance. This review summarizes the recent developments and applications of flexible HDMEAs in closed-loop BMI systems. It delves into the various challenges encountered in the development of ideal flexible HDMEAs for closed-loop BMI systems and highlights the latest methodologies and breakthroughs to address these challenges. These insights could be instrumental in guiding the creation of future generations of flexible HDMEAs, specifically tailored for use in closed-loop BMIs. The review thoroughly explores both the current state and prospects of these advanced arrays, emphasizing their potential in enhancing BMI technology.

A flexible electrode Array for genetic transfection of different layers of the retina by electroporation.

Electroporation (in which the permeability of a cell membrane is increased transiently by exposure to an appropriate electric field) has exhibited great potential of becoming an alternative to adeno-associated virus (AAV)-based retina gene delivery. Electroporation eliminates the safety concerns of employing exogenous viruses and exceeds the limit of AAV cargo size. Unfortunately, several concerns (e.g., relatively high electroporation voltage, poor surgical operability and a lack of spatial selectivity of retina tissue) have prevented electroporation from being approved for clinical application (or even clinical trials). In this study, a flexible micro-electrode array for retina electroporation (FERE) was developed for retina electroporation. A suitably shaped flexible substrate and well-placed micro-electrodes were designed to adapt to the retina curvature and generate an evenly distributed electric field on the retina with a significantly reduced electroporation voltage of 5 V. The FERE provided (for the first time) a capability of controlled gene delivery to the different structural layers of retina tissue by precise control of the distribution of the electrical field. After ensuring the surgical operability of the FERE on rabbit eyeballs, the FERE was verified to be capable of transfecting different layers of retina tissue with satisfactory efficiency and minimum damage. Our method bridges the technical gap between laboratory validation and clinical use of retina electroporation.

Coaxial Electrohydrodynamic Printing of Microscale Core-Shell Conductive Features for Integrated Fabrication of Flexible Transparent Electronics.

Reliable insulation of microscale conductive features is required to fabricate functional multilayer circuits or flexible electronics for providing specific physical/chemical/electrical protection. However, the existing strategies commonly rely on manual assembling processes or multiple microfabrication processes, which is time-consuming and a great challenge for the fabrication of flexible transparent electronics with microscale features and ultrathin thickness. Here, we present a novel coaxial electrohydrodynamic (CEHD) printing strategy for the one-step fabrication of microscale flexible electronics with conductive materials at the core and insulating material at the outer layer. A finite element analysis (FEA) method is established to simulate the CEHD printing process. The extrusion sequence of the conductive and insulating materials during the CEHD printing process shows little effect on the morphology of the core-shell filaments, which can be achieved on different flexible substrates with a minimum conductive line width of 32 ± 3.2 μm, a total thickness of 53.6 ± 4.8 μm, and a conductivity of 0.23 × 107 S/m. The thin insulating layer can provide the inner conductive filament enough protection in 3D, which endows the resultant microscale core-shell electronics with good electrical stability when working in different chemical solvent solutions or under large deformation conditions. Moreover, the presented CEHD printing strategy offers a unique capability to sequentially fabricate an insulating layer, core-shell conductive pattern, and exposed electrodes by simply controlling the material extrusion sequence. The resultant large-area transparent electronics with two-layer core-shell patterns exhibit a high transmittance of 98% and excellent electrothermal performance. The CEHD-printed flexible microelectrode array is successfully used to record the electrical signals of beating mouse hearts. It can also be used to fabricate large-area flexible capacitive sensors to accurately measure the periodical pressure force. We envision that the present CEHD printing strategy can provide a promising tool to fabricate complex three-dimensional electronics with microscale resolution, high flexibility, and multiple functionalities.

Fabrication and characterization of flexible microelectrode arrays for the long-term recording of mammalian brain slices

Abstract A profound understanding of brain disorders is essential for developing more effective treatments, targeted therapies, and reliable diagnoses. Electrical in vitro long-term and repetitive measurements give insight to neural activity of more complex neural circuits and, thus, into malfunctioning caused by illness. Flexible microelectrode arrays (flexMEAs) enable the recording of neural activity with promising spatial and temporal resolution over longer durations. The present study aims to combine the long-term recording by flexMEAs with the cultivation of brain slices inside an incubator. An 8 μm-thin flexMEA with up to 256 electrodes with 30 μm in diameter and interelectrode spacing of 200 μm was fabricated. Characterization showed uniform and reproducible electronic characteristics. Coating with PEDOT:PSS by electropolymerization formed stable low-impedance spectra. The platform is ready for future experiments with brain slices inside an incubator.

Development of a Flexible and Wearable Microelectrode Array Patch Using a Screen-Printed Masking Technique for Accelerated Wound Healing

Development of a Flexible and Wearable Microelectrode Array Patch Using a Screen-Printed Masking Technique for Accelerated Wound Healing

A mosquito mouthpart-like bionic neural probe

Advancements in microscale electrode technology have revolutionized the field of neuroscience and clinical applications by offering high temporal and spatial resolution of recording and stimulation. Flexible neural probes, with their mechanical compliance to brain tissue, have been shown to be superior to rigid devices in terms of stability and longevity in chronic recordings. Shuttle devices are commonly used to assist flexible probe implantation; however, the protective membrane of the brain still makes penetration difficult. Hidden damage to brain vessels during implantation is a significant risk. Inspired by the anatomy of the mosquito mouthparts, we present a biomimetic neuroprobe system that integrates high-sensitivity sensors with a high-fidelity multichannel flexible electrode array. This customizable system achieves distributed and minimally invasive implantation across brain regions. Most importantly, the system’s nonvisual monitoring capability provides an early warning detection for intracranial soft tissues, such as vessels, reducing the potential for injury during implantation. The neural probe system demonstrates exceptional sensitivity and adaptability to environmental stimuli, as well as outstanding performance in postoperative and chronic recordings. These findings suggest that our biomimetic neural-probe device offers promising potential for future applications in neuroscience and brain-machine interfaces.A mosquito mouthpart-like bionic neural probe consisting of a highly sensitive tactile sensor module, a flexible microelectrode array, and implanted modules that mimic the structure of mosquito mouthparts. The system enables distributed implantation of electrode arrays across multiple brain regions while making the implantation minimally invasive and avoiding additional dural removal. The tactile sensor array can monitor the implantation process to achieve early warning of vascular damage. The excellent postoperative short-term recording performance and long-term neural activity tracking ability demonstrate that the system is a promising tool in the field of brain-computer interfaces.

Flexible microelectrode array probe for simultaneous detection of neural discharge and dopamine in striatum of mice aversion system

Simultaneous detection of spike firing and neurotransmitters in mice is conducive to further understanding the mechanism of innate fear induced by aversive stimuli. As the key brain region of the dopamine aversion system, the synchronous changes of neural discharge and neurotransmitter concentration in the striatum under innate fear are less studied. This paper proposed a fabrication method for dual-mode high-sensitive flexible microelectrode array (MEA) probe based on Parylene. The microelectrode sites were modified with platinum black nanoparticles and conductive polymer PEDOT, which can realize single neuron action potential recording and dopamine concentration detection. The electrode sensitivity for dopamine detection can reach 162.3 pA/μM. We used this flexible MEA probe to simultaneously real-time detect electrophysiology activities and dopamine in the striatum of awake mice under aversive stimulation. The results showed that after fear, with the decrease of the mean activity of mice behavior, the action potential firing rate, local field potential (LFP) power in low-frequency (0–30 Hz), and dopamine concentration in the striatum of mice decreased synchronously. The spike firing rate decreased by about 32 %, and the dopamine concentration decreased by about 23.5 %. Our electrodes provide a powerful tool for multimodal neural information detection and our research provides new evidence for striatum involvement in the dopamine aversion system.

Zwitterionic Polymer Coated and Aptamer Functionalized Flexible Micro-Electrode Arrays for In Vivo Cocaine Sensing and Electrophysiology.

The number of people aged 12 years and older using illicit drugs reached 59.3 million in 2020, among which 5.2 million are cocaine users based on the national data. In order to fully understand cocaine addiction and develop effective therapies, a tool is needed to reliably measure real-time cocaine concentration and neural activity in different regions of the brain with high spatial and temporal resolution. Integrated biochemical sensing devices based upon flexible microelectrode arrays (MEA) have emerged as a powerful tool for such purposes; however, MEAs suffer from undesired biofouling and inflammatory reactions, while those with immobilized biologic sensing elements experience additional failures due to biomolecule degradation. Aptasensors are powerful tools for building highly selective sensors for analytes that have been difficult to detect. In this work, DNA aptamer-based electrochemical cocaine sensors were integrated on flexible MEAs and protected with an antifouling zwitterionic poly (sulfobetaine methacrylate) (PSB) coating, in order to prevent sensors from biofouling and degradation by the host tissue. In vitro experiments showed that without the PSB coating, both adsorption of plasma protein albumin and exposure to DNase-1 enzyme have detrimental effects on sensor performance, decreasing signal amplitude and the sensitivity of the sensors. Albumin adsorption caused a 44.4% sensitivity loss, and DNase-1 exposure for 24 hr resulted in a 57.2% sensitivity reduction. The PSB coating successfully protected sensors from albumin fouling and DNase-1 enzyme digestion. In vivo tests showed that the PSB coated MEA aptasensors can detect repeated cocaine infusions in the brain for 3 hrs after implantation without sensitivity degradation. Additionally, the same MEAs can record electrophysiological signals at different tissue depths simultaneously. This novel flexible MEA with integrated cocaine sensors can serve as a valuable tool for understanding the mechanisms of cocaine addiction, while the PSB coating technology can be generalized to improve all implantable devices suffering from biofouling and inflammatory host responses.

Soft Neural Interfacing based on Implantable Graphene Fiber Microelectrode Arrays.

Microelectrode Arrays (MEAs) neural interfaces are considered implantable devices that interact with the nervous system to monitor and/or modulate brain activity. Graphene-based materials are utilized to address some of the current challenges in neural interface design due to their desirable features, such as high conductance, large surface-to-volume ratio, suitable electrochemical properties, biocompatibility, flexibility, and ease of production. In the current study, we fabricated and characterized a type of flexible, ultrasmall, and implantable neurostimulator based on graphene fibers. In this procedure, wet-spinning was employed to create graphene fibers with diameters of 10 to 50 µm. A 10-channel polyimide Printed Circuit Board (PCB) was then custom-designed and manufactured. The fibers were attached to each channel by conductive glue and also insulated by soaking them in a polyurethane solution. The tips were subsequently exposed using a blowtorch. Microstructural information on the fibers was obtained using Scanning Electron Microscopy (SEM), and the measurements of Electrochemical Impedance Spectroscopy (EIS) were conducted for each electrode. Flexible MEAs were created using graphene fibers with diameters ranging from 10 to 50 microns with a spacing of 150 microns. This method leads to producing electrode arrays with any size of fibers and a variety of channel numbers. The flexible neural prostheses can replace conventional electrodes in both neuroscience and biomedical research.

A flexible protruding microelectrode array for neural interfacing in bioelectronic medicine

Recording neural signals from delicate autonomic nerves is a challenging task that requires the development of a low-invasive neural interface with highly selective, micrometer-sized electrodes. This paper reports on the development of a three-dimensional (3D) protruding thin-film microelectrode array (MEA), which is intended to be used for recording low-amplitude neural signals from pelvic nervous structures by penetrating the nerves transversely to reduce the distance to the axons. Cylindrical gold pillars (Ø 20 or 50 µm, ~60 µm height) were fabricated on a micromachined polyimide substrate in an electroplating process. Their sidewalls were insulated with parylene C, and their tips were optionally modified by wet etching and/or the application of a titanium nitride (TiN) coating. The microelectrodes modified by these combined techniques exhibited low impedances (~7 kΩ at 1 kHz for Ø 50 µm microelectrode with the exposed surface area of ~5000 µm²) and low intrinsic noise levels. Their functionalities were evaluated in an ex vivo pilot study with mouse retinae, in which spontaneous neuronal spikes were recorded with amplitudes of up to 66 µV. This novel process strategy for fabricating flexible, 3D neural interfaces with low-impedance microelectrodes has the potential to selectively record neural signals from not only delicate structures such as retinal cells but also autonomic nerves with improved signal quality to study neural circuits and develop stimulation strategies in bioelectronic medicine, e.g., for the control of vital digestive functions.

Manufacturing Processes of Implantable Microelectrode Array for In Vivo Neural Electrophysiological Recordings and Stimulation: A State-Of-the-Art Review.

Electrophysiological recording and stimulation of neuron activities are important for us to understand the function and dysfunction of the nervous system. To record/stimulate neuron activities as voltage fluctuation extracellularly, microelectrode array (MEA) implants are a promising tool to provide high temporal and spatial resolution for neuroscience studies and medical treatments. The design configuration and recording capabilities of the MEAs have evolved dramatically since their invention and manufacturing process development has been a key driving force for such advancement. Over the past decade, since the White House Brain Research Through Advancing Innovative Neurotechnologies (BRAIN) Initiative launched in 2013, advanced manufacturing processes have enabled advanced MEAs with increased channel count and density, access to more brain areas, more reliable chronic performance, as well as minimal invasiveness and tissue reaction. In this state-of-the-art review paper, three major types of electrophysiological recording MEAs widely used nowadays, namely, microwire-based, silicon-based, and flexible MEAs are introduced and discussed. Conventional design and manufacturing processes and materials used for each type are elaborated, followed by a review of further development and recent advances in manufacturing technologies and the enabling new designs and capabilities. The review concludes with a discussion on potential future directions of manufacturing process development to enable the long-term goal of large-scale high-density brain-wide chronic recordings in freely moving animals.

A silk-based self-adaptive flexible opto-electro neural probe

The combination of optogenetics and electrophysiological recording enables high-precision bidirectional interactions between neural interfaces and neural circuits, which provides a promising approach for the study of progressive neurophysiological phenomena. Opto-electrophysiological neural probes with sufficient flexibility and biocompatibility are desirable to match the low mechanical stiffness of brain tissue for chronic reliable performance. However, lack of rigidity poses challenges for the accurate implantation of flexible neural probes with less invasiveness. Herein, we report a hybrid probe (Silk-Optrode) consisting of a silk protein optical fiber and multiple flexible microelectrode arrays. The Silk-Optrode can be accurately inserted into the brain and perform synchronized optogenetic stimulation and multichannel recording in freely behaving animals. Silk plays an important role due to its high transparency, excellent biocompatibility, and mechanical controllability. Through the hydration of the silk optical fiber, the Silk-Optrode probe enables itself to actively adapt to the environment after implantation and reduce its own mechanical stiffness to implant into the brain with high fidelity while maintaining mechanical compliance with the surrounding tissue. The probes with 128 recording channels can detect high-yield well-isolated single units while performing intracranial light stimulation with low optical losses, surpassing previous work of a similar type. Two months of post-surgery results suggested that as-reported Silk-Optrode probes exhibit better implant-neural interfaces with less immunoreactive glial responses and tissue lesions.A silk optical fiber-based Silk-Optrode probe consisting of a natural silk optical fiber and a flexible micro/nano electrode array is reported. The multifunctional soft probe can modify its own Young’s modulus through hydration to achieve accurate implantation into the brain. The low optical loss and single-unit recording abilities allow simultaneous optogenetic stimulation and multichannel readout, which expands the applications in the operation and parsing of neural circuits in behavioral animals.

Spatially controlled, bipolar, cortical stimulation with high-capacitance, mechanically flexible subdural surface microelectrode arrays.

Most neuromodulation approaches rely on extracellular electrical stimulation with penetrating electrodes at the cost of cortical damage. Surface electrodes, in contrast, are much less invasive but are challenged by the lack of proximity to axonal processes, leading to poor resolution. Here, we demonstrate that high-density (40-μm pitch), high-capacitance (>1 nF), single neuronal resolution PEDOT:PSS electrodes can be programmed to shape the charge injection front selectively at depths approaching 300 micrometers with a lateral resolution better than 100 micrometers. These electrodes, patterned on thin-film parylene substrate, can be subdurally implanted and adhere to the pial surface in chronic settings. By leveraging surface arrays that are optically transparent with PEDOT:PSS local interconnects and integrated with depth electrodes, we are able to combine surface stimulation and recording with calcium imaging and depth recording to demonstrate these spatial limits of bidirectional communication with pyramidal neurons in mouse visual cortex both laterally and at depth from the surface.

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.

Nanocone-Array-Based Platinum-Iridium Oxide Neural Microelectrodes: Structure, Electrochemistry, Durability and Biocompatibility Study.

Neural interfaces provide a window for bio-signal modulation and recording with the assistance of neural microelectrodes. However, shrinking the size of electrodes results in high electrochemical impedance and low capacitance, thus limiting the stimulation/recording efficiency. In order to achieve critical stability and low power consumption, here, nanocone-shaped platinum (Pt) with an extensive surface area is proposed as an adhesive layer on a bare Pt substrate, followed by the deposition of a thin layer of iridium oxide (IrOx) to fabricate high-performance nanocone-array-based Pt-IrOx neural microelectrodes (200 μm in diameter). A uniform nanocone-shaped Pt with significant roughness is created via controlling the ratio of NH4+ and Pt4+ ions in the electrolyte, which can be widely applicable for batch production on multichannel flexible microelectrode arrays (fMEAs) and various substrates with different dimensions. The Pt-IrOx nanocomposite-coated microelectrode presents a significantly low impedance down to 0.72 ± 0.04 Ω cm2 at 1 kHz (reduction of ~92.95%). The cathodic charge storage capacity (CSCc) and charge injection capacity (CIC) reaches up to 52.44 ± 2.53 mC cm−2 and 4.39 ± 0.36 mC cm−2, respectively. Moreover, superior chronic stability and biocompatibility are also observed. The modified microelectrodes significantly enhance the adhesion of microglia, the major immune cells in the central nervous system. Therefore, such a coating strategy presents great potential for biomedical and other practical applications.

A flexible implantable microelectrode array for recording electrocorticography signals from rodents.

Electrocorticography signals, the intracranial recording of electrical signatures of the brain, are recorded by non-penetrating planar electrode arrays placed on the cortical surface. Flexible electrode arrays minimize the tissue damage upon implantation. This work shows the design and development of a 32-channel flexible microelectrode array to record electrocorticography signals from the rat's brain. The array was fabricated on a biocompatible flexible polyimide substrate. A titanium/gold layer was patterned as electrodes, and a thin polyimide layer was used for insulation. The fabricated microelectrode array was mounted on the exposed somatosensory cortex of the right hemisphere of a rat after craniotomy and incision of the dura. The signals were recorded using OpenBCI Cyton Daisy Biosensing Boards. The array faithfully recorded the baseline electrocorticography signals, the induced epileptic activities after applying a convulsant, and the recovered baseline signals after applying an antiepileptic drug. The signals recorded by such fabricated microelectrode array from anesthetized rats demonstrate its potential to monitor electrical signatures corresponding to epilepsy. Finally, the time-frequency analyses highlight the difference in spatiotemporal features of baseline and evoked epileptic discharges.

Acute in vivo Recording with a Generic Parylene Microelectrode Array Implanted with Dip-coating Method into the Rat Brain.

Flexible polymer-based microelectrode arrays (MEAs) can reduce tissue inflammation and foreign body response and greatly prolong the lifetime of neural implants. However, standard and customized polymer devices are only accessible to limited groups. To better promote the development and application of polymer MEAs, we have launched the Polymer Implantable Electrode (PIE) Foundry and developed a 64-channel Parylene C-based MEA with generic electrodes layout that can be used to record from both cortical and sub-cortical regions in rodents. In addition, a practical dip-coating protocol for the insertion of the flexible standard Parylene MEA is developed.

High-performance Flexible Microelectrode Array with PEDOT:PSS Coated 3D Micro-cones for Electromyographic Recording.

High signal-to-noise ratio (SNR) electromyography (EMG) recordings are essential for identifying and analyzing single motor unit activity. While high-density electrodes allow for greater spatial resolution, the smaller electrode area translates to a higher impedance and lower SNR. In this study, we developed an implantable and flexible 3D microelectrode array (MEA) with low impedance that enables high-quality EMG recording. With polyimide micro-cones realized by standard photolithography process and PEDOT:PSS coating, this design can increase effective surface area by up to 250% and significantly improve electrical performance for electrode sites with various geometric surface areas, where the electrode impedance is at most improved by 99.3%. Acute EMG activity from mice was recorded by implanting the electrodes in vivo, and we were able to detect multiple individual motor units simultaneously and with high resolution ([Formula: see text]). The charge storage capacity was measured to be 34.2 mC/cm2, indicating suitability of the electrodes for stimulation applications as well.

Distributed implantation of a flexible microelectrode array for neural recording

Flexible multichannel electrode arrays (fMEAs) with multiple filaments can be flexibly implanted in various patterns. It is necessary to develop a method for implanting the fMEA in different locations and at various depths based on the recording demands. This study proposed a strategy for reducing the microelectrode volume with integrated packaging. An implantation system was developed specifically for semiautomatic distributed implantation. The feasibility and convenience of the fMEA and implantation platform were verified in rodents. The acute and chronic recording results provied the effectiveness of the packaging and implantation methods. These methods could provide a novel strategy for developing fMEAs with more filaments and recording sites to measure functional interactions across multiple brain regions.

Flexible Multichannel Neural Probe Developed by Electropolymerization for Localized Stimulation and Sensing

AbstractMiniaturization and minimization of mechanical mismatch in neural probes have been two well‐proven directions in suppressing immune response and improving spatial resolution for neuronal stimulations and recordings. While the high impedance brought by the miniaturization of electrodes has been addressed by using conductive polymers coatings in multiple reports, the stiffness of such coatings remains orders of magnitude higher than that of the brain tissue. Here, a flat neural probe based on a highly flexible microelectrode array with electrodeposited hydrogel coatings poly(2‐hydroxyethyl methacrylate) (pHEMA) and conductive polymer poly(3,4‐ethylenedioxythiophene) polystyrene sulfonate (PEDOT/PSS), with a cross‐section area at only 300 µm × 2.5 µm is presented. The PEDOT/PSS coating provides a low interfacial impedance, and the pHEMA deposition bridges the mechanical mismatch between the probe and the brain tissues. The two layers of polymers modification enhance the signal‐to‐noise ratio and allow the microelectrodes array to be engineered for both recording and stimulation purposes. Besides, in vivo testing of microelectrode arrays implanted in rat hippocampus confirms a high senstivity in neural signal recording and excellent charge injection capacity which can induce long‐term potentiation in neural activities in the hippocampus. The probes provide a robust and low‐cost solution to the brain interfaces problem.