Graphene Microelectrode Research Articles

Graphene Microelectrode Arrays, 4D Structured Illumination Microscopy, and a Machine Learning Spike Sorting Algorithm Permit the Analysis of Ultrastructural Neuronal Changes During Neuronal Signaling in a Model of Niemann-Pick Disease Type C.

Simultaneously recording network activity and ultrastructural changes of the synapse is essential for advancing understanding of the basis of neuronal functions. However, the rapid millisecond-scale fluctuations in neuronal activity and the subtle sub-diffraction resolution changes of synaptic morphology pose significant challenges to this endeavor. Here, specially designed graphene microelectrode arrays (G-MEAs) are used, which are compatible with high spatial resolution imaging across various scales as well as permit high temporal resolution electrophysiological recordings to address these challenges. Furthermore, alongside G-MEAs, an easy-to-implement machine learning algorithm is developed to efficiently process the large datasets collected from MEA recordings. It is demonstrated that the combined use of G-MEAs, machine learning (ML) spike analysis, and 4D structured illumination microscopy (SIM) enables monitoring the impact of disease progression on hippocampal neurons which are treated with an intracellular cholesterol transport inhibitor mimicking Niemann-Pick disease type C (NPC), and show that synaptic boutons, compared to untreated controls, significantly increase in size, leading to a loss in neuronal signaling capacity.

Introducing phosphoric acid to fluorinated polyimide towards high performance laser induced graphene electrodes for high energy micro-supercapacitors

Micro-supercapacitors (MSCs) have wide application prospects in microelectronic fields such as wearable electronics due to merits of stable performance, high safety and easy integration. However, the relatively low energy density of MSCs limits their practical application. In this context, phosphorus and fluorine co-doped laser-induced graphene (FP-LIG) microelectrodes were fabricated from fluorinated polyimide containing phosphoric acid by laser direct writing (LDW) method. The introduced phosphoric acid slows down the decomposition of –CF3 during the LDW process, resulting in much more ordered and stable pores; meanwhile, phosphorus entered the graphene lattice to replace some carbon atoms, forming a C3PO structure, which not only stabilizes the interface between the electrode and the electrolyte and therefore achieves an enlarged working potential of 1.4 V, but also increases the wettability of the electrode. Using FP-3-LIG microelectrodes and PVA/H2SO4 as the gel electrolyte, the assembled FP-3-MSC demonstrates significantly enhanced energy density, delivering an energy density of 10.40 μWh cm−2 (@0.09 mA cm−2), 2.7 times that of F-MSC and 346.7 times that of MSC. FP-3-MSC has excellent cyclic stability, displaying an areal capacitance retention rate of above 90 % after 10,000 long cycles. In addition, FP-3-MSC demonstrates excellent flexibility, indicating promising potential in the field of flexible wearable electronics.

Detection of Dopamine Based on Aptamer-Modified Graphene Microelectrode.

In this paper, a novel aptamer-modified nitrogen-doped graphene microelectrode (Apt-Au-N-RGOF) was fabricated and used to specifically identify and detect dopamine (DA). During the synthetic process, gold nanoparticles were loaded onto the active sites of nitrogen-doped graphene fibers. Then, aptamers were modified on the microelectrode depending on Au-S bonds to prepare Apt-Au-N-RGOF. The prepared microelectrode can specifically identify DA, avoiding interference with other molecules and improving its selectivity. Compared with the N-RGOF microelectrode, the Apt-Au-N-RGOF microelectrode exhibited higher sensitivity, a lower detection limit (0.5 μM), and a wider linear range (1~100 μM) and could be applied in electrochemical analysis fields.

Honeycomb-Patterned Graphene Microelectrodes: A Promising Approach for Safe and Effective Retinal Stimulation Based on Electro-Thermo-Mechanical Modeling and Simulation.

The main objective of the present study is to use graphene as electrode neural interface material to design novel microelectrodes topology for retinal prosthesis and investigate device operation safety based on the computational framework. The study's first part establishes the electrode material selection based on electrochemical impedance and the equivalent circuit model. The second part of the study is modeling at the microelectrode-tissue level to investigate the potential distribution, generated resistive heat dissipation, and thermally induced stress in the tissue due to electrical stimulation. The formulation of Joule heating and thermal expansion between microelectrode-tissue-interface employing finite element method modeling is based on the three coupled equations, specifically Ohm's law, Navier's equation, and Fourier equation. Electrochemical simulation results of electrode material reveal that single-layer and few-layer graphene-based microelectrode has a specific impedance in the range of 0.02- [Formula: see text], comparable to platinum counterparts. The microelectrode of [Formula: see text] size can stimulate retinal tissue with a threshold current in the range of 8.7- [Formula: see text]. Such stimulation with the observed microelectrode size indicates that both microelectrodes and retinal tissue stay structurally intact, and the device is thermally and mechanically stable, functioning within the safety limit. The results reveal the viability of high-density graphene-based microelectrodes for improved interface as stimulating electrodes to acquire higher visual acuity. Furthermore, the novel microelectrodes design configuration in the honeycomb pattern gives the retinal tissue non-invasive heating and minimal stress upon electrical stimulation. Thus, it paves the path to designing a graphene-based microelectrode array for retinal prosthesis for further in vitro or in vivo studies.

High-performance micro supercapacitor assembled by laser-induced graphene electrode and hydrogel electrolyte with excellent interfacial wettability for high capacitance

In this work, we demonstrate a facile, rational and novel strategy to assemble micro-supercapacitors (MSCs) via employing laser-induced graphene (LIG) microelectrodes and sodium alginate/polyacrylamide hydrogel electrolytes soaked in sulfuric acid solution (SA/PAAM-H2SO4). The microelectrodes are conductive graphene materials with porous structure that use laser direct writing technology to carbonize insulating polyimide (PI) sheets. The importance of good wetting of the electrode material by the electrolyte on the performance of the device is explained. There is good compatibility and interfacial contact between electrode and electrolyte. Impressively, the SA/PAAM-H2SO4 hydrogel electrolyte with high ionic conductivity (574.7 mS cm−1) also exhibit good water retention capability and performs normally even after 60 h of exposure to air. The capacitance of LIG-MSCs based on SA/PAAM-H2SO4 hydrogel is comparable to that of PVA/H2SO4 hydrogel. The LIG-MSCs exhibit high areal capacitance of 7.12 mF cm−2 and outstanding cycling stability. Moreover, our LIG-MSCs displays excellent mechanical flexibility and stable performance, the capacitance is almost unaffected even in highly bent states and during continuously hammering. Furthermore, the LIG-MSCs can be arbitrarily connected in series and in parallel without the requirement of metal-based interconnects for high-voltage and high-capacitance output. Our design strategy aims to provide new insights for the development of flexible MSCs.

Dual-Mode Graphene Field-Effect Transistor Biosensor with Isothermal Nucleic Acid Amplification.

Despite a substantial increase in testing facilities during the pandemic, access remains a major obstacle, particularly in low-resource and remote areas. This constraint emphasizes the need for high-throughput potential point-of-care diagnostic tools in environments with limited resources. Loop-mediated isothermal amplification (LAMP) is a promising technique, but improvements in sensitivity are needed for accurate detection, especially in scenarios where the virus is present in low quantities. To achieve this objective, we present a highly sensitive detection approach of a dual-mode graphene-based field-effect transistor (G-FET) biosensor with LAMP. The G-FET biosensor, which has a transparent graphene microelectrode array on a glass substrate, detects LAMP products in less than 30 min using both observable color changes and Dirac point voltage measurements, even in samples with low viral concentrations. This dual-mode G-FET biosensor emerges as a potential alternative to conventional RT-PCR for severe acute respiratory syndrome-associated coronavirus (SARS-CoV)-2 detection or point-of-care testing, particularly in resource-constrained scenarios such as developing countries. Moreover, its capacity for colorimetric detection with the naked eye enhances its applicability in diverse settings.

High-density transparent graphene arrays for predicting cellular calcium activity at depth from surface potential recordings.

Optically transparent neural microelectrodes have facilitated simultaneous electrophysiological recordings from the brain surface with the optical imaging and stimulation of neural activity. A remaining challenge is to scale down the electrode dimensions to the single-cell size and increase the density to record neural activity with high spatial resolution across large areas to capture nonlinear neural dynamics. Here we developed transparent graphene microelectrodes with ultrasmall openings and a large, transparent recording area without any gold extensions in the field of view with high-density microelectrode arrays up to 256 channels. We used platinum nanoparticles to overcome the quantum capacitance limit of graphene and to scale down the microelectrode diameter to 20 µm. An interlayer-doped double-layer graphene was introduced to prevent open-circuit failures. We conducted multimodal experiments, combining the recordings of cortical potentials of microelectrode arrays with two-photon calcium imaging of the mouse visual cortex. Our results revealed that visually evoked responses are spatially localized for high-frequency bands, particularly for the multiunit activity band. The multiunit activity power was found to be correlated with cellular calcium activity. Leveraging this, we employed dimensionality reduction techniques and neural networks to demonstrate that single-cell and average calcium activities can be decoded from surface potentials recorded by high-density transparent graphene arrays.

A label-free H1N1 influenza virus immunosensor based on an N-LIG/Au laser induced graphene microelectrode.

A label-free immunosensor based on an N-doped laser direct graphene (N-LIG)/Au electrode was proposed for H1N1 influenza virus detection. By utilizing the instantaneous high temperature of laser irradiation, N atoms are generated by the decomposition of melamine dripped onto the surface of an LIG electrode to obtain N-LIG with higher conductivity. The doping of N atoms provides a large number of active sites for LIG microelectrodes. Combined with the electrodeposition of Au NPs, and covalently crosslinking antibodies, a simple, highly sensitive and stable immunosensing interface is constructed. The proposed H1N1 influenza virus immunosensor has a detection range of 0.01 fg mL-1 to 10 ng mL-1 with a detection limit as low as 0.004 fg mL-1. The constructed sensor has ultra-high sensitivity and good selectivity and can be used for complex biological sample analysis, with potential application prospects in preventing the large-scale spread of influenza. Taking advantage of N-LIG electrode's properties will provide opportunities for developing portable electrochemical biosensors for health and environmental applications.

Preparation of self-supporting vertically/horizontally grown graphene microelectrodes for neurotransmitter determination

The development of microelectrodes for the rapid in situ detection of neurotransmitters and their metabolic levels in human biofluids has considerable significance in biomedical research. In this study, self-supported graphene microelectrodes with B-doped, N-doped, and B- and N-co-doped vertical graphene (BVG, NVG, and BNVG, respectively) nanosheets grown on horizontal graphene (HG) were fabricated for the first time. The high electrochemical catalytic activity of BVG/HG on monoamine compounds was explored by investigating the influence of B and N atoms and the VG layer thickness on the response current of neurotransmitters. Quantitative analysis using the BVG/HG electrode in a blood-like environment with pH 7.4 indicated that the linear concentration ranges were 1–400 and 1–350 μM for dopamine (DA) and serotonin (5-HT), with limits of detection (LODs) of 0.271 and 0.361 μM, respectively. For tryptophan (Trp), the sensor measured a wide linear concentration range of 3–1500 μM over a wide pH range of 5.0–9.0, with the LOD fluctuating between 0.58 and 1.04 μM. Furthermore, the BVG/HG microelectrodes could be developed as needle- and pen-type sensors for the detection of DA, 5-HT, and Trp in human blood and gastrointestinal secretion samples.

A graphene microelectrode array based microfluidic device for in situ continuous monitoring of biofilms.

In situ continuous monitoring of bacterial biofilms has been a challenging job so far, but it is fundamental to the screening of novel anti-biofilm reagents. In this work, a microfluidic system utilizing a graphene-modified microelectrode array sensor was proposed to realize the dynamic state of bacterial biofilm monitoring by electrochemical impedance. The results illustrated that the observation window period of the biofilm state is significantly prolonged due to the increment of bacterial cell load on the sensing interface, thereby greatly improving the sensing signal quality. Simulation of anti-biofilm drug screening demonstrated that the performance of this method manifestly exceeded that of its endpoint counterparts.

I‐VISION project. A new step in artificial vision

AbstractThe i‐VISION project aims to develop next‐generation of retinal prosthesis technology in efforts to restore high‐acuity artificial vision to individuals blinded by retinal degeneration. Retinal prostheses operate by electrically stimulating retinal neurons that survive the degeneration in a spatiotemporal pattern that is interpreted by the brain as vision. Presently, the challenge to implement thousands of small‐diameter electrodes and increasingly more sophisticated stimulation protocols that can adapt to the patients' needs has not been met, which severely limits the quality of restored vision in current retinal prostheses.In this presentation I will present how i‐VISION is set to address some of the challenges hindering the performance of current retinal prostheses, with a focus on the use of revolutionary miniaturized electrode nanotechnology based on graphene. Graphene microelectrodes are capable of mediating high‐charge injection highly localized that will improve the spatial resolution at the neural interface, and locally monitoring the stimulation efficacy, allowing a future adaptive therapy strategy. In addition, I will present work of the i‐VISION consortium on the use of advanced in vitro and in vivo imaging techniques of the evoked retinal and cortical responses to quantify the area of neurons activated by the electrical stimulation, with the ultimate goal of generating maps of retina‐visual cortex interconnectivity.

E-Cannula reveals anatomical diversity in sharp-wave ripples as a driver for the recruitment of distinct hippocampal assemblies.

The hippocampus plays a critical role in spatial navigation and episodic memory. However, research on invivo hippocampal activity dynamics mostly relies on single modalities, such as electrical recordings or optical imaging, with respectively limited spatial and temporal resolution. Here, we develop the E-Cannula, integrating fully transparent graphene microelectrodes with imaging cannula, which enables simultaneous electrical recording and two-photon calcium imaging from the exact same neural populations across an anatomically extended region of the mouse hippocampal CA1 stably across several days. The large-scale multimodal recordings show that sharp wave ripples (SWRs) exhibit spatiotemporal wave patterns along multiple axes in two-dimensional (2D) space with different spatial extents and temporal propagation modes. Notably, distinct SWR wave patterns are associated with the selective recruitment of orthogonal CA1 cell assemblies. These results demonstrate the utility of the E-Cannula as a versatile neurotechnology with the potential for future integration with other optical components.

Spatial-Interleaving Graphene Supercapacitor with High Area Energy Density and Mechanical Flexibility.

The booming portable electronics market has raised huge demands for the development of supercapacitors with mechanical flexibility and high power density in the finite area; however, this is still unsatisfied by the currently thickness-confined sandwich design or the in-plane interdigital configuration with limited mechanical features. Here, a spatial-interleaving supercapacitor (SI-SC) is first designed and constructed, in which the graphene microelectrodes are reversely stacked layer by layer within a three-dimensional (3D) space. Because each microelectrode matches well with four counter microelectrodes and all 3D spatial-interleaving microelectrodes have narrow interspaces that maintain the efficient ions transport in the whole device, this SI-SC has a prominent liner capacitance increase along with the device thickness. As a result, the high specific areal capacitance of 36.46 mF cm-2 and 5.34 μWh cm-2 energy density is achieved on the 100 μm thick device. Especially, the microelectrodes in each layer are interdigitated, ensuring the outstanding mechanical flexibility of SI-SC, with ∼98.7% performance retention after 104 cycles of bending tests, realizing the excellent integration of high area energy density and mechanical flexibility in the finite area. Furthermore, the SI-SC units can be easily integrated into wearable electronics to power wristwatches, light-emitting diodes (LEDs), calculators, and so on for practical applications.

Electrochemical Detection of a Few Copies of Unamplified SARS-CoV-2 Nucleic Acids by a Self-Actuated Molecular System.

The existing electrochemical biosensors lack controllable and intelligent merit to modulate the sensing process upon external stimulus, leading to challenges in analyzing a few copies of biomarkers in unamplified samples. Here, we present a self-actuated molecular-electrochemical system that consists of a tentacle and a trunk modification on a graphene microelectrode. The tentacle that contains a probe and an electrochemical label keeps an upright orientation, which increases recognition efficiency while decreasing the pseudosignal. Once the nucleic acids are recognized, the tentacles nearby along with the labels are spontaneously actuated downward, generating electrochemical responses under square wave voltammetry. Thus, it detects unamplified SARS-CoV-2 RNAs within 1 min down to 4 copies in 80 μL, 2–6 orders of magnitude lower than those of other electrochemical assays. Double-blind testing and 10-in-1 pooled testing of nasopharyngeal samples yield high overall agreement with reverse transcription-polymerase chain reaction results. We fabricate a portable prototype based on this system, showing great potential for future applications.

Graphene-Enabled High-Performance Electrokinetic Focusing and Sensing.

Transverse isoelectric focusing, i.e., isoelectric focusing that is normal to the fluid-flow direction, is an electrokinetic method ideal for micro total analysis. However, a major challenge remains: There is no electrode system integrable in a microfluidic device to allow reliable transverse isoelectric focusing and electrokinetic sensing. Here, we overcome this barrier by developing devices that incorporate microelectrodes made of monolayer graphene. We find that the electrolysis stability over time for graphene microelectrodes is >103× improved compared to typical microfabricated inert-metal microelectrodes. Through transverse isoelectric focusing between graphene microelectrodes, within minutes, specific proteins can be separated and concentrated to scales of ∼100 μm. Based on the concentrating effect and the high optical transparency of graphene, we develop a three-dimensional multistream microfluidic strategy for label-free detection of the proteins at same processing position with a sensitivity that is ∼102× higher than those of the state-of-the-art label-free sensors. These results demonstrate the advantage of monolayer-graphene microelectrodes for high-performance electrokinetic analysis to allow lab-on-a-chips of maximal time and size efficiencies.

Minute-sensitive real-time monitoring of neural cells through printed graphene microelectrodes

Real-time and high-throughput cytometric monitoring of neural cells exposed to injury mechanisms is invaluable for in-vitro studies. Electrical impedance spectroscopy via microelectrode arrays is a label-free technique for monitoring of neural growth and their detachment upon death. In this method, the interface material plays a vital role to provide desirable attachment cues for the cell network. Thus, here we demonstrate the electrohydrodynamic patterning of aqueous graphene for microelectrode fabrication. We investigated whether the wrinkled surface morphology of the electrodes fabricated by this deposition method expands their electroactive surface area and thus enables a rapid response time. The nano-scale quality of the graphene lattice is characterized by Raman spectroscopy and Transmittance electron microscopy. N27 rat dopaminergic neural cells were cultured on the chips and the surface morphology of the microelectrodes during cellular growth was investigated by Scanning electrode spectroscopy. Attachment of the neural population on the graphene microelectrodes was parametrized and the change in the impedance spectrum of this cell population was quantified at 10 Hz to 10 kHz frequencies along with the change in TUBB3 gene expression. The viability test of the cell population on the biosensor demonstrated no significant difference in comparison to the control, and a cell density of 2289 cell/mm2 was achieved. As a proof of concept, the confluent N27 cell population was exposed to UV and its cytotoxic impact on neural detachment and lift-off was monitored. The multiplexed detection of cellular activity was reported with a temporal resolution of one minute.

Graphene Microelectrodes for Real-Time Impedance Spectroscopy of Neural Cells.

Understanding the changes in the electrochemical properties of neural cells upon exposure to stress factors imparts vital information about the conditions prior to their death. This study presents a graphene-based biosensor for real-time monitoring of N27 rat dopaminergic neural cells which characterizes cell adhesion and cytotoxicity factors through impedance spectroscopy. The aim was to monitor the growth of the entire cell network via a nonmetallic flexible electrode array. Therefore, a water-based graphene solution was formulized as a conductive ink, 3D-printed into a flexible substrate through an electrohydrodynamic approach, resulting in electrodes with a conductivity of 6750 s/m. The presented high-throughput method enabled microscale monitoring of the entire cell network via the design of PDMS-based growth channels. The electrical resistance of the cell network was measured continuously along with their network density, constituting a mean density of 1890 cell/mm2 at full cell confluency. The results demonstrate the applicability of the impedance-based sensing of the cell network for rapid screening of the cytotoxic elements, and the real-time effect of UV exposure on dopaminergic neural cells was reported as an immediate application of the device.

Green Flexible Graphene–Inorganic‐Hybrid Micro‐Supercapacitors Made of Fallen Leaves Enabled by Ultrafast Laser Pulses

AbstractThe development of green flexible micro‐supercapacitors (MSCs) is one of the biggest challenges in future wearable electronics. Flexible MSCs are mainly produced from non‐biodegradable synthetic polymers, resulting in massive electronic waste. Moreover, complex multi‐step fabrication increases their production cost. Here, the direct fabrication of highly conductive, intrinsically flexible, and green microelectrodes from naturally fallen leaves in ambient air using femtosecond laser pulses without any additional materials is reported. Hierarchically porous graphene is patterned on different types of leaves via a facile, mask‐less, scalable, and one‐step laser writing. Leaves consist of biominerals, which decompose into inorganic crystals that serve as nucleation sites for the growth of 3D mesoporous few‐layer graphene. The femtosecond laser‐induced graphene (FsLIG) microelectrodes formed on leaves have lower sheet resistance (23.3 Ω sq−1) than their synthetic polymer counterparts and exhibit an outstanding areal capacitance (34.68 mF cm−2 at 5 mV s−1) and capacitance retention (≈99% after 50 000 charge/discharge cycles). The FsLIG MSCs on a single leaf could easily power a light‐emitting diode or a table clock and could be applied in wearable electronics, smart houses, and Internet of Things.

Laser direct write of heteroatom-doped graphene on molecularly controlled polyimides for electrochemical biosensors with nanomolar sensitivity

Fabrication of heteroatom-doped graphene electrodes remains a challenging endeavor, especially on flexible substrates. Precise chemical and morphological control is even more challenging for patterned microelectrodes. We herein demonstrate a scalable process for directly generating micropatterns of heteroatom-doped porous graphene on polyimide with different backbones using a continuous-wave infrared laser. Conventional two-step polycondensation of 4,4′-oxydianiline with three different tetracarboxylic dianhydrides enabled the fabrication of fully aromatic polyimides with various internal linkages such as phenylene, trifluoromethyl or sulfone groups. Accordingly, we leverage this laser-induced polymer-to-doped-graphene conversion for fabricating electrically conductive microelectrodes with efficient utilization of heteroatoms (N-doped, F-doped, and S-doped). Tuning laser fluence enabled achieving electrical resistivity lower than ∼13 Ω sq−1 for F-doped and N-doped graphene. Finally, our microelectrodes exhibit superior performance for electrochemical sensing of dopamine, one of the important neurotransmitters in the brain. Compared with carbon fiber microelectrodes, the gold standard in electrochemical dopamine sensing, our F-doped high surface area graphene microelectrodes demonstrated 3 order of magnitude higher sensitivity per unit area, detecting dopamine concentrations as low as 10 nM with excellent reproducibility. Hence, our approach is promising for facile fabrication of microelectrodes with superior capabilities for various electrochemical and sensing applications including early diagnosis of neurological disorders.

3D fuzzy graphene microelectrode array for dopamine sensing at sub-cellular spatial resolution

The development of a high sensitivity real-time sensor for multi-site detection of dopamine (DA) with high spatial and temporal resolution is of fundamental importance to study the complex spatial and temporal pattern of DA dynamics in the brain, thus improving the understanding and treatments of neurological and neuropsychiatric disorders. In response to this need, here we present high surface area out-of-plane grown three-dimensional (3D) fuzzy graphene (3DFG) microelectrode arrays (MEAs) for highly selective, sensitive, and stable DA electrochemical sensing. 3DFG microelectrodes present a remarkable sensitivity to DA (2.12 ± 0.05 nA/nM, with LOD of 364.44 ± 8.65 pM), the highest reported for nanocarbon MEAs using Fast Scan Cyclic Voltammetry (FSCV). The high surface area of 3DFG allows for miniaturization of electrode down to 2 × 2 μm2, without compromising the electrochemical performance. Moreover, 3DFG MEAs are electrochemically stable under 7.2 million scans of continuous FSCV cycling, present exceptional selectivity over the most common interferents in vitro with minimum fouling by electrochemical byproducts and can discriminate DA and serotonin (5-HT) in response to the injection of their 50:50 mixture. These results highlight the potential of 3DFG MEAs as a promising platform for FSCV based multi-site detection of DA with high sensitivity, selectivity, and spatial resolution.