An Opto-electrophysiology Neural Probe with Photoelectric Artifact-Free for Advanced Single-Neuron Analysis.

Opto-electrophysiology neural probes targeting single-cell levels offer an important avenue for elucidating the intrinsic mechanisms of the nervous system using different physical quantities, representing a significant future direction for brain-computer interface (BCI) devices. However, the highly integrated structure poses significant challenges to fabrication processes and the presence of photoelectric artifacts complicates the extraction and analysis of target signals. Here, we propose a highly miniaturized and integrated opto-electrophysiology neural probe for electrical recording and optical stimulation at the single-cell/subcellular level. The design of a total internal reflection layer addresses the photoelectric artifacts that are more pronounced in single-cell devices compared to conventional implantable BCI devices. Finite element simulations and electrical signal tests demonstrate that the opto-electrophysiology neural probe eliminates the photoelectric artifacts in the time domain, which represents a significant breakthrough for optoelectrical integrated BCI devices. Our proposed opto-electrophysiology neural probe holds substantial potential for promoting the development of in vivo BCI devices and developing advanced therapeutic strategies for neurological disorders.

In situ self-referenced intracellular two-electrode system for enhanced accuracy in single-cell analysis.

Since the emergence of single-cell electroanalysis, the two-electrode system has become the predominant electrochemical system for real-time behavioral analysis of single-cell and multicellular populations. However, due to the transmembrane placement of the two electrodes, cellular activities can be interrupted by the transmembrane potentials, and the test results are susceptible to influences from factors such as intracellular solution, membrane, and bulk solution. These limitations impede the advancement of single-cell analysis. Here, we propose a highly miniaturized and integrated in situ self-referenced intracellular two-electrode system (IS-SRITES), wherein both the working and reference electrodes are positioned inside the cell. Additionally, we demonstrated the stability (0.28 mV/h) of the solid-contact in situ Ag/AgCl reference electrode and the ability of the system to conduct standard electrochemical testing in a wide pH range (pH 6.0-8.0). Cell experiments confirmed the non-destructive performance of the electrode system towards cells and its capacity for real-time monitoring of intra- and extracellular pH values. Moreover, through equivalent circuits, finite element simulations, and drug delivery experiments, we illustrated that the IS-SRITES can yield more accurate test results and exhibit enhanced resistance to interference from the extracellular environment. Our proposed system holds the potential to enable the precise detection of intracellular substances and optimize the existing model of the electrode system for intracellular signal detection, thereby spearheading advancements in single-cell analysis.

Multifunctional IrOx Neural Probe for In Situ Dynamic Brain Hypoxia Evaluation.

Perioperative cerebral hypoxia and neonatal hypoxia-ischemic encephalopathy are the main triggers that lead to temporary or permanent brain dysfunction. The pathogenesis is intimately correlated to neural activities and the pH of the microenvironment, which calls for a high demand for in situ multitype physiological signal acquisition in the brain. However, conventional pH sensing neural interfaces cannot obtain the characteristics of multimodes, multichannels, and high spatial resolution of physiological signals simultaneously. Here, we report a multifunctional implantable iridium oxide (IrOx) neural probe (MIIONP) combined with electrophysiology recording, in situ pH sensing, and neural stimulation for real-time dynamic brain hypoxia evaluation. The neural probe modified with IrOx films exhibits outstanding electrophysiology recording and neural stimulation performance and long-term stable high spatial pH sensing resolution of about 100 µm, and the cytotoxicity of IrOx microelectrodes was investigated as well. In addition, 4 weeks' tracking of the same neuron firing and instantaneous population spike captured during electrical stimulation was achieved by MIIONP. Finally, in a mouse brain hypoxia model, the MIIONP has demonstrated the capability of synchronous in situ recording of the pH and neural firing changes in the brain, which has a valuable application in dynamic brain disease evaluation through real-time acquisition of multiple physiological signals.

Brainmask: an ultrasoft and moist micro-electrocorticography electrode for accurate positioning and long-lasting recordings.

Bacterial cellulose (BC), a natural biomaterial synthesized by bacteria, has a unique structure of a cellulose nanofiber-weaved three-dimensional reticulated network. BC films can be ultrasoft with sufficient mechanical strength, strong water absorption and moisture retention and have been widely used in facial masks. These films have the potential to be applied to implantable neural interfaces due to their conformality and moisture, which are two critical issues for traditional polymer or silicone electrodes. In this work, we propose a micro-electrocorticography (micro-ECoG) electrode named "Brainmask", which comprises a BC film as the substrate and separated multichannel parylene-C microelectrodes bonded on the top surface. Brainmask can not only guarantee the precise position of microelectrode sites attached to any nonplanar epidural surface but also improve the long-lasting signal quality during acute implantation with an exposed cranial window for at least one hour, as well as the in vivo recording validated for one week. This novel ultrasoft and moist device stands as a next-generation neural interface regardless of complex surface or time of duration.

A flexible neural implant with ultrathin substrate for low-invasive brain-computer interface applications.

Implantable brain-computer interface (BCI) devices are an effective tool to decipher fundamental brain mechanisms and treat neural diseases. However, traditional neural implants with rigid or bulky cross-sections cause trauma and decrease the quality of the neuronal signal. Here, we propose a MEMS-fabricated flexible interface device for BCI applications. The microdevice with a thin film substrate can be readily reduced to submicron scale for low-invasive implantation. An elaborate silicon shuttle with an improved structure is designed to reliably implant the flexible device into brain tissue. The flexible substrate is temporarily bonded to the silicon shuttle by polyethylene glycol. On the flexible substrate, eight electrodes with different diameters are distributed evenly for local field potential and neural spike recording, both of which are modified by Pt-black to enhance the charge storage capacity and reduce the impedance. The mechanical and electrochemical characteristics of this interface were investigated in vitro. In vivo, the small cross-section of the device promises reduced trauma, and the neuronal signals can still be recorded one month after implantation, demonstrating the promise of this kind of flexible BCI device as a low-invasive tool for brain-computer communication.

Dense Packed Drivable Optrode Array for Precise Optical Stimulation and Neural Recording in Multiple-Brain Regions.

The input-output function of neural networks is complicated due to the huge number of neurons and synapses, and some high-density implantable electrophysiology recording tools with a plane structure have been developed for neural circuit studies in recent years. However, traditional plane probes are limited by the record-only function and inability to monitor multiple-brain regions simultaneously, and the complete cognition of neural networks still has a long way away. Herein, we develop a three-dimensional (3D) high-density drivable optrode array for multiple-brain recording and precise optical stimulation simultaneously. The optrode array contains four-layer probes with 1024 microelectrodes and two thinned optical fibers assembled into a 3D-printed drivable module. The recording performance of microelectrodes is optimized by electrochemical modification, and precise implantation depth control of drivable optrodes is verified in agar. Moreover, in vivo experiments indicate neural activities from CA1 and dentate gyrus regions are monitored, and a tracking of the neuron firing for 2 weeks is achieved. The suppression of neuron firing by blue light has been realized through high-density optrodes during optogenetics experiments. With the feature of large-scale recording, optoelectronic integration, and 3D assembly, the high-density drivable optrode array possesses an important value in the research of brain diseases and neural networks.

Self-adaptive cardiac optogenetics device based on negative stretching-resistive strain sensor.

Precision medicine calls for high demand of continuous, closed-loop physiological monitoring and accurate control, especially for cardiovascular diseases. Cardiac optogenetics is promising for its superiority of cell selectivity and high time-space accuracy, but the efficacy of optogenetics relative to the input of light stimulus is detected and controlled separately by discrete instruments in vitro, which suffers from time retardation, energy consumption, and poor portability. Thus, a highly integrated system based on implantable sensors combining closed-loop self-monitoring with simultaneous treatment is highly desired. Here, we report a self-adaptive cardiac optogenetics system based on an original negative stretching-resistive strain sensor array for closed-loop heart rate recording and self-adaptive light intensity control. The strain sensor exhibits a dual and synchronous capability of precise monitor and physiological-electrical-optical regulation. In an in vivo ventricular tachycardia model, our system demonstrates the potential of a negative stretching-resistive device in controlling-in-sensor electronics for wearable/implantable autodiagnosis and telehealth applications.

Fabrication and Characterization of Iridium Oxide pH Microelectrodes Based on Sputter Deposition Method.

pH value plays an important role in many fields such as chemistry and biology; therefore, rapid and accurate pH measurement is very important. Because of its advantages in preparation, wide test range, rapid response, and good biocompatibility, iridium oxide material has received more and more attention. In this paper, we present a method for preparing iridium oxide pH microelectrodes based on the sputter deposition method. The sputtering parameters of iridium oxide are also studied and optimized. Open-circuit potential tests show that microelectrodes exhibit near-Nernstian pH response with good linearity (about 60 mV/pH), fast response, high stability (a slight periodic fluctuation of potential change <2.5 mV in 24 h), and good reversibility in the pH range of 1.00-13.00.

An artefact-resist optrode with internal shielding structure for low-noise neural modulation.

OBJECTIVE: The combination of optical manipulation of neural activities with electrophysiology recording is a promising technology for discovering mechanisms of brain disorders and mapping brain networks. However, fiber-based optrode is limited by the large size of light source and the winding of optical fiber, which hinders animal's natural movement. Meanwhile, the laser diode (LD)-based optrode restricted to the stimulation-locked artefacts will contaminate neural signal acquired from recording channels. APPROACH: Here, a reformative low-noise optrode with internal grounded shielding layer is proposed to mitigate the stimulus-locked artefacts generated during LDactivation for the application of optogenetics. MAIN RESULTS: The artefact mitigation capacity of grounded shielding was verified via simulation and experiments with transient amplitude of artefacts declined from over 5 mV to approximately 200 µV in-vitro. Meanwhile, the stimulation parameters were used based on previous studies by which neurons were activated without over heating the tissue as characterized by in-vitro studies (the output optical intensity is 823 ± 38 mW mm-2). Furthermore, the microelectrodes were modified with Poly (3, 4-ethylenedioxythiophene)-poly (styrenesulfonate) (PEDOT: PSS) to increase the signal recording quality of the optrode. Finally, in-vivo optogenetics experiments were carried in the hippocampus of one mouse and the results showed our low-noise optrode was qualified to achieve high-quality neural recording (signal-to-noise ratio about 13) and specific neuron stimulation simultaneously. SIGNIFICANCE: These results suggest the low-noise optrodes exhibit the ability of manipulating and recording neural dynamics and they are excellent candidates for neuroscience research.

Flexible and stretchable opto-electric neural interface for low-noise electrocorticogram recordings and neuromodulation in vivo.

Optogenetic-based neuromodulation tools is evolving for the basic neuroscience research in animals combining optical manipulation and electrophysiological recordings. However, current opto-electric integrated devices attaching on cerebral cortex for electrocorticogram (ECoG) still exist potential damage risks for both brain tissue and electrode, due to the mechanical mismatch and brain deformation. Here, we propose a stretchable opto-electric integrated neural interface by integrating serpentine-shaped electrodes and multisite micro-LEDs onto a hyperelastic substrate, as well as a serpentine-shaped metal shielding embedded in recording electrode for low-noise signal acquisition. The delicate structure design, ultrasoft encapsulation and independent fabrication followed by assembly are beneficial to the conformality, reliability and yield. In vitro accelerated deterioration and reciprocating tensile have demonstrated good performance and high stability. In vivo optogenetic activation of focal cortical areas of awaked mouse expressing Channelrhodopsin-2 is realized with simultaneous high-quality recording. We highlight the potential use of this multifunctional neural interface for neural applications.

The use of a double-layer platinum black-conducting polymer coating for improvement of neural recording and mitigation of photoelectric artifact.

The impedance of electrode and photostimulation artifacts (short-duration and high-amplitude spikes) are still hindering the employment of silicon-based neural probe in optogenetics. A fiber-based optrode modified with a double-layer platinum black-poly (3,4ethylenedioxythiophene) PEDOT/poly (4-styrenesulfonate) PSS (Pt-PP) coating has been developed for improvement of neural recording quality and mitigation of photoelectric artifact simultaneously. The Pt-PP coating was made by layer-by-layer electrochemical deposition followed by the ultrasonication and Cyclic Voltammetry (CV) scanning to verify its mechanical and electrochemical stability. Both in-vitro and in-vivo experiments demonstrated that Pt-PP coated optrode had outstanding recording performance (high signal-to-noise ratio about 9.64) and low photoelectric amplitude (850 µV). The artifact recovery time of Pt-PP coated optrode (0.3 ms) after photostimulation was significantly decreased when compared to platinum black (6 ms) or PEDOT/PSS (0.7 ms) coated one which has potential to retain high-quality neural signals in animal experiments. At last, the optogenetics experiments revealed the capability of Pt-PP coated optrode to record the change in neural spike rate with certain spatial resolution and shorter artifact recovery time. These results suggest that Pt-PP coating has great potential for neural electrodes in the application of neuroscience.

Flexible bioelectrodes with enhanced wrinkle microstructures for reliable electrochemical modification and neuromodulation in vivo.

Limited electrode size with high electrochemical performance and reliability of modified materials are two of the main concerns for flexible neural electrodes in recent years. Here, an effective fabrication method of enhanced micro-scale wrinkles based on oil-pretreated hyperelastic substrates (PDMS and Ecoflex) is proposed for the application of microelectrode biosensors. Compared to pre-stretching or compressing methods, this approach has better advantages including compatibility with MEMS processes on wafer and easy replication. Wrinkled gold microelectrodes exhibit superior electrochemical properties than the flat one, and no crack or delamination occurs after electroplating PEDOT:PSS and platinum black on wrinkled microelectrodes. Cyclic voltammetry (CV) scanning for 2500 times is performed to investigate adhesion and stability of modified materials. For the modified microelectrodes, no significant change is observed in charge storage capacity (CSC) and impedance at 1 kHz, whereas PEDOT:PSS coated flat microelectrodes appears delamination. Ultrasonication and cycling forces are also conducted on modified microelectrodes, which demonstrates little influence on the wrinkled ones. Flexible wrinkled microelectrodes are further verified by in-vivo ECoG recordings combined with optogenetics in mice. These results highlight the importance of micro-structure in neural electrode design and tremendous application potentials in flexible electronics.

Three-dimensional drivable optrode array for high-resolution neural stimulations and recordings in multiple brain regions.

The brain-computer interface (BCI) devices are of prime important for study of nervous system as well as diagnosis and treatment of neurological disorders. To meet the needs of the BCI devices in high-density integration and multi-functionalization, 3-dimensional (3D) drivable optrode array with laser diodes (LDs) coupled waveguides was developed. The unique device realizes the 3D integration of the optrodes and avoids fiber tangle and tissue heating by adopting LD coupled waveguide structure. Besides, the postoperative position adjustment of the optrode array was achieved by integrating with a 3D printed micro-drive. Most importantly, high-resolution neural stimulations and recordings were achieved for study of working memory related neural circuits in four brain regions of mice including prelimbic cortex (PrL), mediodorsal thalamic nucleus (MD), dorsal medial caudate nucleus (dmCP) and posterior motor cortex 2 (pM2). The results indicate that this novel device is promising for the research of complex neural networks.