Fabrication Of Microelectrodes Research Articles

Experimental investigations and optimization of non-electrical parameters during microelectrode fabrication

Experimental investigations and optimization of non-electrical parameters during microelectrode fabrication

General In Situ Engineering of Carbon-Based Materials on Carbon Fiber for In Vivo Neurochemical Sensing.

Developing real-time, dynamic, and in situ analytical methods with high spatial and temporal resolutions is crucial for exploring biochemical processes in the brain. Although in vivo electrochemical methods based on carbon fiber (CF) microelectrodes are effective in monitoring neurochemical dynamics during physiological and pathological processes, complex post modification hinders large-scale productions and widespread neuroscience applications. Herein, we develop a general strategy for the in situ engineering of carbon-based materials to mass-produce functional CFs by introducing polydopamine to anchor zeolitic imidazolate frameworks as precursors, followed by one-step pyrolysis. This strategy demonstrates exceptional universality and design flexibility, overcoming complex post-modification procedures and avoiding the delamination of the modification layer. This simplifies the fabrication and integration of functional CF-based microelectrodes. Moreover, we design highly stable and selective H+, O2, and ascorbate microsensors and monitor the influence of CO2 exposure on the O2 content of the cerebral tissue during physiological and ischemia-reperfusion pathological processes.

Bessel beam fabrication of graphitic micro electrodes in diamond using laser bursts

We present the fabrication of conductive graphitic microelectrodes in diamond by using pulsed Bessel beams in the burst mode laser writing regime. The graphitic wires are created in the bulk of a 500 μm thick monocrystalline HPHT diamond (with (100) orientation) perpendicular to the sample surface, without beam scanning or sample translation. In particular, the role of different burst features in the resistivity of such electrodes is investigated for two very different sub-pulse durations namely 200 fs and 10 ps, together with the role of thermal annealing. Micro-Raman spectroscopy is implemented to investigate the laser-induced crystalline modification, and the results obtained by using two different laser repetition rates, namely 20 Hz and 200 kHz, are compared. A comparison of the micro-Raman spectra and of the resistivity of the electrodes fabricated respectively with 10 ps single pulses and with bursts (of sub-pulses) of similar total duration has also been made, and we show that the burst mode writing regime allows to fabricate more conductive micro electrodes, thanks to the heat accumulation process leading to stronger graphitization. Moreover, the microfabrication of diamond by means of the longest available bursts (~ 46.7 ps duration) featured by 32 sub-pulses of 200 fs duration, with intra-burst time delay of 1.5 ps (sub-THz bursts), leads to graphitic wires with the lowest resistivity values obtained in this work, especially at low repetition rate such as 20 Hz. Indeed, micro electrodes with resistivity on the order of 0.01 Ω cm can be fabricated by Bessel beams in the burst mode regime even when the bursts are constituted by femtosecond laser sub-pulses, in contrast with the results of the standard writing regime with single fs pulses typically leading to less conductive micro electrodes.

A PEDOT: PSS/GO fiber microelectrode fabricated by microfluidic spinning for dopamine detection in human serum and PC12 cells.

Rapid and accurate in situ determinationof dopamine is of great significance in the study of neurological diseases. In this work, poly (3,4-ethylenedioxythiophene): poly (styrenesulfonic acid) (PEDOT: PSS)/graphene oxide (GO) fibers were fabricated by an effective method based on microfluidic wet spinning technology. The composite microfibers with stratified and dense arrangement were continuously prepared by injecting PEDOT: PSS and GO dispersion solutions into a microfluidic chip. PEDOT: PSS/GO fiber microelectrodes with high electrochemical activity and enhanced electrochemical oxidation activity of dopamine were constructed by controlling the structure composition of the microfibers with varying flow rate. The fabricated fiber microelectrode had a low detection limit (4.56 nM) and wide detection range (0.01-8.0 µM) for dopamine detection with excellent stability, repeatability, and reproducibility. In addition, the PEDOT: PSS/GO fiber microelectrode prepared was successfully used for the detection of dopamine in human serum and PC12 cells. The strategy for the fabrication of multi-component fiber microelectrodes isa new and effective approach for monitoring the intercellular neurotransmitter dopamine and has highpotential as an implantable neural microelectrode.

Assessment of chemotherapeutic effects on cancer cells using adhesion noise spectroscopy.

With cancer as one of the leading causes of death worldwide, there is a need for the development of accurate, cost-effective, easy-to-use, and fast drug-testing assays. While the NCI 60 cell-line screening as the gold standard is based on a colorimetric assay, monitoring cells electrically constitutes a label-free and non-invasive tool to assess the cytotoxic effects of a chemotherapeutic treatment on cancer cells. For decades, impedance-based cellular assays extensively investigated various cell characteristics affected by drug treatment but lack spatiotemporal resolution. With progress in microelectrode fabrication, high-density Complementary Metal Oxide Semiconductor (CMOS)-based microelectrode arrays (MEAs) with subcellular resolution and time-continuous recording capability emerged as a potent alternative. In this article, we present a new cell adhesion noise (CAN)-based electrical imaging technique to expand CMOS MEA cell-biology applications: CAN spectroscopy enables drug screening quantification with single-cell spatial resolution. The chemotherapeutic agent 5-Fluorouracil exerts a cytotoxic effect on colorectal cancer (CRC) cells hampering cell proliferation and lowering cell viability. For proof-of-concept, we found sufficient accuracy and reproducibility for CAN spectroscopy compared to a commercially available standard colorimetric biological assay. This label-free, non-invasive, and fast electrical imaging technique complements standardized cancer screening methods with significant advances over established impedance-based approaches.

Fabrication of pyrolytic carbon interdigitated microelectrodes by maskless UV photolithography with epoxy-based photoresists SU-8 and mr-DWL

Maskless UV photolithography is increasingly used, especially in research environments where low turn-around time for new designs improves productivity. Here, we fabricate pyrolytic carbon interdigitated microelectrodes with small interelectrode gaps, good adhesion to the carrier substrate, high surface area and excellent electrochemical properties using maskless UV photolithography with two negative epoxy-based photoresists, namely the commonly used SU-8 and the recently developed mr-DWL. The minimum realizable trench width in 15 μm thick photoresist films is 2.4 ± 0.15 μm for mr-DWL 5 and 3.1 ± 0.10 μm for SU-8 2035. After pyrolysis, the two resulting pyrolytic carbon materials show similar electrochemical properties. However, shrinkage during pyrolysis is significantly lower for mr-DWL compared to SU-8, which is beneficial for the fabrication of interdigitated microelectrodes. Furthermore, delamination of the electrodes during processing and operation is prevented due to the introduction of poly silicon adhesion structures. This work provides valuable insights into maskless UV lithography as well as into the pyrolytic carbon process to increase the yield, performance and productivity for fabrication of microelectrodes.

Fabrication of Microelectrodes, Suction Electrodes, and Focal Electrodes for Electrophysiological Recording in Drosophila.

Electrophysiological recording is a group of techniques used to record electrical field potentials generated by cells. These techniques rely on several types of electrodes, which can be manufactured in the laboratory. In intracellular recording, glass microelectrodes are used to pierce the cell membrane, and then to measure the electrical potential difference between the inside and the outside of the cell. Another technique, called loose patch or focal recording, is similar to intracellular recording but the electrode tip does not pierce into the cell membrane. Rather, the electrode tip is placed near a nerve or the postsynaptic side of the neuromuscular junction (NMJ) to record extracellular changes in local potentials. A third technique involves a suction electrode, which is used to draw part of the motor nerve into the electrode so that electrical pulses can be applied to elicit action potentials of the nerve. Suction electrodes are specifically used to evoke synaptic potentials at the Drosophila larval NMJ. This protocol details some basic methods for manufacturing microelectrodes used for intracellular recording and two-electrode voltage-clamp and loose patch electrodes used for focal recording. In addition, a method is provided for manufacturing homemade suction electrodes used for nerve stimulation.

High areal energy zinc-ion micro-batteries enabled by 3D printing

Rechargeable zinc-ion micro-batteries (ZIMBs) are promising candidates for powering various microelectronic devices, although they typically exhibit low areal energy of ∼0.1 mWh cm−2. In this study, we reported the design and assembly of zinc micro-batteries through an ink extrusion-based printing approach. In the process, a three-axis printer was adopted, allowing for the fabrication of microelectrodes with large areal loading and diverse morphologies. Vanadium oxide anchored with carbon nanotubes was employed as the cathode material, zinc powder was used as the anode material, and a polyacrylamide gel containing ethylene glycol was selected as the electrolyte. The printed micro-batteries demonstrate an areal capacity of 0.51 mAh cm−2 and an energy density of 0.37 mWh cm−2 at a charge-discharge current of 0.2 mA, outperforming state-of-the-art zinc counterparts. This work provides a simple yet efficient solution to the development of high-energy micro-batteries.

Nanoporous graphene-based thin-film microelectrodes for in vivo high-resolution neural recording and stimulation

One of the critical factors determining the performance of neural interfaces is the electrode material used to establish electrical communication with the neural tissue, which needs to meet strict electrical, electrochemical, mechanical, biological and microfabrication compatibility requirements. This work presents a nanoporous graphene-based thin-film technology and its engineering to form flexible neural interfaces. The developed technology allows the fabrication of small microelectrodes (25 µm diameter) while achieving low impedance (∼25 kΩ) and high charge injection (3–5 mC cm−2). In vivo brain recording performance assessed in rodents reveals high-fidelity recordings (signal-to-noise ratio >10 dB for local field potentials), while stimulation performance assessed with an intrafascicular implant demonstrates low current thresholds (<100 µA) and high selectivity (>0.8) for activating subsets of axons within the rat sciatic nerve innervating tibialis anterior and plantar interosseous muscles. Furthermore, the tissue biocompatibility of the devices was validated by chronic epicortical (12 week) and intraneural (8 week) implantation. This work describes a graphene-based thin-film microelectrode technology and demonstrates its potential for high-precision and high-resolution neural interfacing.

Fabrication of a Palladium Hydride pH Microelectrode with an Extended Lifetime

Stable miniaturized pH microelectrodes are crucial in exploring localized pH changes, but most suffer from instability and short lifetimes. Palladium hydride in a mixer phase has been reported to measure the solution pH taking advantage of the interaction between the alloyed hydrogen atoms and hydrogen ions in the solution. However, with the escape of the hydrogen atoms from the palladium lattice, the palladium hydride will transition to single phase and become invalid for pH measurement. The duration of the mixer-phase palladium hydride is even shorter as the sensor is made small. A method to stabilize the mixer-phase is needed for the development of palladium hydride sensors.A method of synthesizing the mixer-phase palladium hydride in dimethylformamide at an elevated temperature has been reported. The synthesized palladium hydride can be stable under aerobic conditions. We used the stable palladium hydride to fabricate a pH microelectrode. In contrast to electrochemically generated palladium hydride microelectrodes, chemically generated palladium hydride microelectrodes are longer lasting and importantly have good an analytical performance under aerobic conditions. Chemically generated palladium hydride microelectrodes perform best in acid to neutral electrolytes devoid of Cl-. They can readily be produced on 10 µm diameter disk platinum microelectrodes, which makes them attractive candidates for future localized electrochemical studies.

Development of multi-depth probing 3D microelectrode array to record electrophysiological activity within neural cultures

Microelectrode arrays (MEAs) play a crucial role in investigating the electrophysiological activities of neuronal populations. Although two-dimensional neuronal cell cultures have predominated in neurophysiology in monitoring in-vitro the electrophysiological activity, recent research shifted toward culture using three-dimensional (3D) neuronal network structures for developing more sophisticated and realistic neuronal models. Nevertheless, many challenges remain in the electrophysiological analysis of 3D neuron cultures, among them the development of robust platforms for investigating the electrophysiological signal at multiple depths of the 3D neurons’ networks. While various 3D MEAs have been developed to probe specific depths within the layered nervous system, the fabrication of microelectrodes with different heights, capable of probing neural activity from the surface as well as from the different layers within the neural construct, remains challenging. This study presents a novel 3D MEA with microelectrodes of different heights, realized through a multi-stage mold-assisted electrodeposition process. Our pioneering platform allows meticulous control over the height of individual microelectrodes as well as the array topology, paving the way for the fabrication of 3D MEAs consisting of electrodes with multiple heights that could be tailored for specific applications and experiments. The device performance was characterized by measuring electrochemical impedance, and noise, and capturing spontaneous electrophysiological activity from neurospheroids derived from human induced pluripotent stem cells. These evaluations unequivocally validated the significant potential of our innovative multi-height 3D MEA as an avant-garde platform for in vitro 3D neuronal studies.

Use of 3D Printing Techniques to Fabricate Implantable Microelectrodes for Electrochemical Detection of Biomarkers in the Early Diagnosis of Cardiovascular and Neurodegenerative Diseases.

This Review provides a comprehensive overview of 3D printing techniques to fabricate implantable microelectrodes for the electrochemical detection of biomarkers in the early diagnosis of cardiovascular and neurodegenerative diseases. Early diagnosis of these diseases is crucial to improving patient outcomes and reducing healthcare systems' burden. Biomarkers serve as measurable indicators of these diseases, and implantable microelectrodes offer a promising tool for their electrochemical detection. Here, we discuss various 3D printing techniques, including stereolithography (SLA), digital light processing (DLP), fused deposition modeling (FDM), selective laser sintering (SLS), and two-photon polymerization (2PP), highlighting their advantages and limitations in microelectrode fabrication. We also explore the materials used in constructing implantable microelectrodes, emphasizing their biocompatibility and biodegradation properties. The principles of electrochemical detection and the types of sensors utilized are examined, with a focus on their applications in detecting biomarkers for cardiovascular and neurodegenerative diseases. Finally, we address the current challenges and future perspectives in the field of 3D-printed implantable microelectrodes, emphasizing their potential for improving early diagnosis and personalized treatment strategies.

Simultaneous local sensing of two chemical properties with dual soft probe scanning electrochemical microscopy

A new method for the rapid and economic fabrication of dual soft microelectrodes for Soft-Probe-Scanning electrochemical microscopy (Soft-Probe-SECM) and their use for the simultaneous local detection of locally generated species is presented. The process is based on encapsulating electrode wires, such as Pt, Au and carbon fiber, by UV-photo-polymerization of a dielectric material leading to two microdisc electrodes, which are individually addressable. Probe stability is realized by using a flexible plastic support. The thickness of the dielectric material that controls the distance between the microelectrodes and the substrate surface during lateral scans over a substrate in contact mode is highly controllable by the probe fabrication parameters. The thickness of the dielectric material of the soft probes fabricated here varies between 27 µm and 35 µm, while the two microelectrodes inter-distance varies between 180 µm and 250 µm. The dual soft probes combine Pt, Au and C microdiscs of different radii (12.5 µm for Pt and Au and 4 µm for C). The flexible selection of the electrode materials enables the fabrication of dual soft probes with electrode materials suitable to detect electro-active analytes of interest at solid/solution interfaces. The dual soft probes can be used in double amperometric, double potentiometric or mixed potentiometric-amperometric modes. As a proof-of concept of the latter mode, a dual platinum black/Pt (PtB/Pt)–iridium oxide/Au (IrOx/Au) soft probe is used to image in real time the chemically induced dissolution of inkjet printed silver patterns in 0.2 mM HNO3, generating locally reactive nitrogen species (RNS) and simultaneously changing the pH. RNS amounts are measured at 0.8 V (vs. Ag/AgCl, KCl, reference electrode) at the PtB/Pt microelectrode, while the corresponding pH changes are obtained from open circuit potential measurements with the IrOx/Au microelectrode.

Wafer-patterned, permeable, and stretchable liquid metal microelectrodes for implantable bioelectronics with chronic biocompatibility.

Implantable bioelectronics provide unprecedented opportunities for real-time and continuous monitoring of physiological signals of living bodies. Most bioelectronics adopt thin-film substrates such as polyimide and polydimethylsiloxane that exhibit high levels of flexibility and stretchability. However, the low permeability and relatively high modulus of these thin films hamper the long-term biocompatibility. In contrast, devices fabricated on porous substrates show the advantages of high permeability but suffer from low patterning density. Here, we report a wafer-scale patternable strategy for the high-resolution fabrication of supersoft, stretchable, and permeable liquid metal microelectrodes (μLMEs). We demonstrate 2-μm patterning capability, or an ultrahigh density of ~75,500 electrodes/cm2, of μLME arrays on a wafer-size (diameter, 100 mm) elastic fiber mat by photolithography. We implant the μLME array as a neural interface for high spatiotemporal mapping and intervention of electrocorticography signals of living rats. The implanted μLMEs have chronic biocompatibility over a period of eight months.

Fabrication of conductive micro electrodes in diamond bulk using pulsed Bessel beams

High-quality, in-bulk conductive graphitic microelectrodes are fabricated perpendicular to the surface of a 500 μm thick monocrystalline CVD diamond sample using pulsed Bessel beams. With a 12o cone angle beam, different pulse parameters are explored to optimize the graphitic wires which are written without sample translation. The quality of the electrodes and their electrical and structural properties have been analysed through current-voltage characterization and micro-Raman spectroscopy. We have found that higher pulse duration favours better conductivity while pulse energy has an optimum value for the same. This trend is confirmed by the presence and the different amounts of graphitic-like sp2 bonded carbon revealed by the micro-Raman spectra in different configurations. Using suitable writing parameters, we are able to create electrodes with the resistivity of 0.04 Ω cm, which, to the best of our knowledge, is one of the lowest values ever reported in literature in the case of graphitic-like wires fabricated through laser micromachining.

Recent Advances in CMOS Electrochemical Biosensor Design for Microbial Monitoring: Review and Design Methodology.

Rapid, high-sensitivity, and real-time characterization of microorganisms plays a significant role in several areas, including clinical diagnosis, human healthcare, early detection of outbreaks, and the protection of living beings. Integrating microbiology and electrical engineering promises the development of low-cost, miniaturized, autonomous, and high-sensitivity sensors to quantify and characterize bacterial strains at various concentrations. Electrochemical-based biosensors are receiving particular attention in microbiological applications among the different biosensing devices. Several approaches have been adopted to design and fabricate cutting-edge, miniaturized, and portable electrochemical biosensors to track and monitor bacterial cultures in real time. These techniques differ in their sensing interface circuits and microelectrode fabrication. The goals of this review are (i) to summarize the current state of CMOS sensing circuit designs in label-free electrochemical biosensors for bacteria monitoring and (ii) to discuss the material and size of the electrodes used in electrochemical biosensors in microbiological applications. In this paper, we reviewed the latest and most advanced CMOS integrated interface circuits that have recently been used in electrochemical biosensors to identify and characterize bacteria species, such as impedance spectroscopy, capacitive, amperometry, and voltammetry, etc. In addition to the interface circuit design, other crucial factors, such as the material and scale of the electrodes, must be considered to increase the sensitivity of electrochemical biosensors. Surveying the literature in this field improves our knowledge about the impact of electrode designs and materials on sensing precision and will help future designers adapt, design, and fabricate appropriate electrode configurations based on their application. Thus, we summarized the conventional microelectrode designs and materials mainly employed in microbial sensors, including interdigitated electrodes (IDEs), microelectrode arrays (MEAs), paper, and carbon-based electrodes, etc.

Iridium Oxides Based Potentiometric Sensor for pH Monitoring in Biological Samples

The fabrication and long-term application of a pH Au microelectrode based on an iridium oxide film are reported. A uniform iridium oxide film with a typical thickness of around 1 µm was coated on the microelectrode surface through a 2-step procedure involving electrodeposition at constant potential and further continuous voltammetric scans. A super-Nernstian slope of around 77 mV per pH unit was found from open circuit potential measurements in a broad pH range (2 to 10) in 0.01 mol L-1 phosphate buffer. It was demonstrated experimentally that the short-term pH precision of the IrOx sensor is ± 0.1 pH. The response stability was maintained in the physiological pH range, and the sensor exhibited excellent reproducibility, long-term stability, and a short response time of &lt; 10 s. The results reported in this work confirmed that iridium oxide showed very promising pH sensing performance and can serve as an electrode material for detecting local pH changes in samples of increased complexity, such as juice fruits, culture medium, synthetic urine, and blood.

Printed Silk Microelectrode Arrays for Electrophysiological Recording and Controlled Drug Delivery.

The use of soft and flexible bioelectronic interfaces can enhance the quality for recording cells' electrical activity by ensuring a continuous and intimate contact with the smooth, curving surfaces found in the physiological environment. This work develops soft microelectrode arrays (MEAs) made of silk fibroin (SF) films for recording interfaces that can also serve as a drug delivery system. Inkjet printing is used as a tool to deposit the substrate, conductive electrode, and insulator, as well as a drug-delivery nanocomposite film. This approach is highly versatile, as shown in the fabrication of carbon microelectrodes, sandwiched between a silk substrate and a silk insulator. The technique permits the development of thin-film devices that can be employed for in vitro extracellular recordings of HL-1 cell action potentials. The tuning of SF by applying an electrical stimulus to produce a permeable layer that can be used in on-demand drug delivery systems is also demonstrated. The multifunctional MEA developed here can pave the way for in vitro drug screening by applying time-resolved and localized chemical stimuli.

Conductive graphitic wires generation in diamond by means of pulsed Bessel beam micromachining

We present the fabrication of transverse graphitic microelectrodes in a 500 μm thick synthetic diamond bulk by means of pulsed Bessel beams. By suitably placing the elongated focal length of the Bessel beam across the entire sample, the graphitic wires grow from the bottom surface up to the top during multiple shot irradiation. The morphology of the microstructures generated and the micro-Raman spectra are studied as a function of the laser parameters and the diamond crystal orientation. We show the possibility to generate high conductivity microelectrodes, which are crucial for the application of electric fields or current transport/collection in various chips and detectors.

Fabrication of ITO microelectrodes and electrode arrays using a low-cost CO2 laser plotter.

Good electronic (Rs = ∼5 Ω sq-1) and optical properties (transmittance: >83%) make indium tin oxide (ITO) an attractive electrode substrate. Despite the commercial availability of high-quality ITO and some low-cost methods for direct deposition being in use by now, the definition of patterns is still a concern. Putting their popularity and extensive use aside, the manufacturing of ITO electrodes so far lacks a rapid, highly reproducible, flexible, cost-effective, easy patterning process that could surpass difficult, time-consuming techniques such as lithography. Herein, we present a low-cost method based on CO2 laser irradiation for preparing ITO microelectrodes and electrode arrays. Electrodes of different sizes and shapes were examined to identify the performance of the proposed methods. Direct ablation of the ITO layer was optimized for rectangular electrodes of 25, 50, and 100 μm in width, while laser cutting of scotch tape stencils and subsequent wet etching were used to create circular electrodes with a diameter of 1.75 mm. A multielectrode array system consisting 8 of these circular electrodes was fabricated on a (25 × 25) mm2 plate, characterized electrochemically through cyclic voltammetry (CV) and scanning electrochemical microscopy (SECM), and as an example application used for monitoring the anchoring behavior of HeLa and HepG2 cell cultures through cell-based electrochemical impedance spectroscopy. Together, the direct ablation method and preparation of laser cut stencils form a complete toolbox, which allows for low-cost and fast fabrication of ITO electrodes for a wide variety of applications. To demonstrate the general availability of the method, we have also prepared a batch of electrodes using a laser plotter in a local printing shop, achieving high intra-workshop reproducibility.