Ultrathin Cell‐Membrane‐Mimic Phosphorylcholine Polymer Film Coating Enables Large Improvements for In Vivo Electrochemical Detection

Resisting biomolecule adsorption onto the surface of brain‐implanted microelectrodes is a key issue for in vivo monitoring of neurochemicals. Herein, we demonstrate that an ultrathin cell‐membrane‐mimic film of ethylenedioxythiophene tailored with zwitterionic phosphorylcholine (EDOT‐PC) electropolymerized onto the surface of a carbon fiber microelectrode (CFE) not only resists protein adsorption but also maintains the sensitivity and time response for in vivo monitoring of dopamine (DA). As a consequence, the as‐prepared PEDOT‐PC/CFEs could be used as a new reliable platform for tracking DA in vivo and would help understand the physiological and pathological functions of DA.     

Micrometer-scale transient ion transport for real-time pH assay in living rat brains

Ion transport has been widely used for various applications such as sensing, desalination and energy conversion; however, nearly all applications are based on steady-state ion transport. Herein, we for the first time demonstrate the capability of transient ion transport for in vivo sensing with both high spatial (∼μm) and temporal (∼ms) resolution by using pH as the model target. Transient ion transport behavior (i.e., time-dependent ion current change) was observed by applying high-frequency pulse potential. Importantly, we proposed the ion distribution transient model for this time-dependent ion transport behavior. With this model, the temporal resolution of the as-developed pH microsensor based on ion current was improved to the ms level, thus satisfying the requirement of neurochemical recording. Moreover, our microsensor features good reproducibility, selectivity, and reversibility, and can thus real-time monitor the pH change in living rat brains. This study demonstrates the first example of in vivo sensing based on ion transport, opening a new way to neurochemical monitoring with ultrahigh spatiotemporal resolution. This study is also helpful to understand the transient process of asymmetric ion transport.

Micrometer-scale transient ion transport has been successfully used for constructing a high spatiotemporal resolution and performance microsensor, which could be used for real-time monitoring the change of pH in rat brains.     

A Generalizable and Noncovalent Strategy for Interfacing Aptamers with a Microelectrode for the Selective Sensing of Neurotransmitters In Vivo

The selective sensing of neurochemicals is essential for understanding the chemical basis of brain function and pathology. Interfacing the excellent recognition features of aptamers with in vivo compatible carbon fiber microelectrode (CFE)-based electroanalytical systems offers a plausible means to achieve this end. However, this is challenging in terms of coupling chemistry, stability, and versatility. Here, we present a new interfacial functionalization strategy based on the assembly of aptamer cholesterol amphiphiles (aptCAs) on the alkyl chain-functionalized CFE. The noncovalent cholesterol-alkyl chain interactions effectively immobilize aptamers onto the CFE surface, allowing the generation of a highly selective system for probing neurochemical dynamics in living systems and opening up a vast array of new opportunities for designing in vivo sensors for exploring brain chemistry.     

Recent progress in improving the performance of in vivo electrochemical microsensor based on materials

In vivo monitoring of neurochemicals is important for exploring the mechanism and function of the central nervous system. In vivo electrochemical microsensor benefiting from high temporal and spatial resolution has been demonstrated to be one effective strategy for neurochemical detection. However, due to the complex biological environment, microsensor faces huge challenges in sensitivity, selectivity, stability, and biocompatibility. Materials with good electron-transfer, rough surface, and easy functionalization are widely used to enhance the performance of microsensor. In this review, we summarize the recent progress in improving the performance of in vivo electrochemical microsensor based on materials.     

Real‐time Monitoring of Discrete Synaptic Release Events and Excitatory Potentials within Self‐reconstructed Neuromuscular Junctions

Chemical synaptic transmission is central to the brain functions. In this regard, real‐time monitoring of chemical synaptic transmission during neuronal communication remains a great challenge. In this work, in vivo‐like oriented neural networks between superior cervical ganglion (SCG) neurons and their effector smooth muscle cells (SMC) were assembled in a microfluidic device. This allowed amperometric detection of individual neurotransmitter release events inside functional SCG‐SMC synapse with carbon fiber nanoelectrodes as well as recording of postsynaptic potential using glass nanopipette electrodes. The high vesicular release activities essentially involved complex events arising from flickering fusion pores as quantitatively established based on simulations. This work allowed for the first time monitoring in situ chemical synaptic transmission under conditions close to those found in vivo, which may yield important and new insights into the nature of neuronal communications.     

Fiber-optic microsensor for high resolution pCO2 sensing in marine environment

A fast responding fiber-optic microsensor for sensing pCO2 in marine sediments with high spatial resolution is presented. The tip diameter varies typically between 20 and 50 microm. In order to make the pH-indicator 8-hydroxypyrene-1,3,6-trisulfonate soluble in the ethyl cellulose matrix, it was lipophilized with tetraoctylammonium as the counterion [HPTS-(TOA)4]. The microsensor was tuned to sense very low levels of dissolved carbon dioxide which are typically present in marine systems. The detection limit is 0.04 hPa pCO2 which corresponds to 60 ppb CO2 of dissolved carbon dioxide. A soluble Teflon derivative with an extraordinarily high gas permeability was chosen as a protective coating to eliminate interferences by ionic species like chloride or pH. Response times of less than 1 min were observed. The performance of the new microsensor is described with respect to reproducibility of the calibration curves, dynamic range, temperature behavior, long term stability and storage stability. The effect of hydrogen sulfide as an interferent, which is frequently present in anaerobic sediment layers, was studied in detail.     

Fiber-optic microsensor for high resolution pCO2 sensing in marine environment

A fast responding fiber-optic microsensor for sensing pCO₂ in marine sediments with high spatial resolution is presented. The tip diameter varies typically between 20 and 50 μm. In order to make the pH-indicator ¶8-hydroxypyrene-1,3,6-trisulfonate soluble in the ethyl cellulose matrix, it was lipophilized with tetraoctylammonium as the counterion [HPTS-(TOA)₄]. The microsensor was tuned to sense very low levels of dissolved carbon dioxide which are typically present in marine systems. The detection limit is 0.04 hPa pCO₂ which corresponds to 60 ppb CO₂ of dissolved carbon dioxide. A soluble Teflon derivative with an extraordinarily high gas permeability was chosen as a protective coating to eliminate interferences by ionic species like chloride or pH. Response times of less than 1 min were observed. The performance of the new microsensor is described with respect to reproducibility of the calibration curves, dynamic range, temperature behavior, long term stability and storage stability. The effect of hydrogen sulfide as an interferent, which is frequently present in anaerobic sediment layers, was studied in detail.     

Overoxidation of carbon-fiber microelectrodes enhances dopamine adsorption and increases sensitivity

The voltammetric responses of carbon-fiber microelectrodes with a 1.0 V and a 1.4 V anodic limit were compared in this study. Fast-scan cyclic voltammetry was used to characterize the response to dopamine and several other neurochemicals. An increase in the adsorption properties of the carbon fiber leads to an increase in sensitivity of 9 fold in vivo. However the temporal response of the sensor is slower with the more positive anodic limit. Increased electron transfer kinetics also causes a decrease in the relative sensitivity for dopamine vs. other neurochemicals, and a change in their cyclic voltammograms. Stimulated release in the caudate-putamen was pharmacologically characterized in vivo using Ro-04-1284 and pargyline, and was consistent with that expected for dopamine.     

Measurement of dioxygen by electrocatalytic reduction on microelectrodes modified with Nafion and methyl viologen

Nafion/methyl viologen (MV) has been chemically modified on a gold disk microelectrode (GDME). The electrochemistry of the Nafion/MV modified GDME is investigated by cyclic voltammetry (CV). Linear sweep voltammetry (LSV) and differential pulse amperometry (DPA) show that the Nafion/MV modified GDME exhibits very high electrocatalytic activity toward dioxygen reduction with good reproducibility and high sensitivity. The electrocatalytic peak current is found to be linear with the dioxygen concentration in the range of 3.44x10(-7) to 2.59x10(-4) mol l(-1) (at 25 degrees C), with a correlation coefficient of 0.9978. The detection limit (signal/noise=3) is calculated to be 0.19 mumol l(-1). The response time of the microsensor for dioxygen measurement is less than 15 s. For ten parallel measurements for 8.50 mumol l(-1) dioxygen, the relative standard deviation (RSD) is found to be 2.7%. The sensitivity of the microsensor is 0.17 nA mumol(-1) l(-1). This microsensor has been successfully employed to measure the concentration of dioxygen in real samples. The quantity of dioxygen, released from the three kinds of chloroplasts of plant leaves under different illumination, is monitored by the Nafion/MV modified gold microsensor. In order to survey the dioxygen concentration in vivo, a Nafion/MV modified carbon fiber microelectrode (CFME) is fabricated by a modification procedure similar to that of the Nafion/MV GDME. As a preliminary test, the dioxygen levels in the different areas of rat brain are determined by the Nafion/MV modified carbon fiber microsensors. The mechanism of the catalytic reaction is also addressed.     

Flexible dopamine-sensing fiber based on potentiometric method for long-term detection in vivo

Achieving real-time, continuous and long-term monitoring of dopamine(DA) in vivo is essential for revealing brain functions and preventing and treating neurogenic diseases. However, it remains challenging to achieve a low limit of detection(LOD) and high neuron-compatibility at the same time for the current microsensors, resulting in the failure of long-term and accurate detection of DA in vivo. A DA-sensing fiber was achieved by the potentiometric method to possess a low LOD of 5 nM, 1-3 orders of magnitude lower than amperometry and differential-pulse voltammetry. The sensing fiber showed a wide linear range from 5 to 185 nM that well matched the DA concentration(26-40 nM) in vivo. After implantation, the sensing fiber showed no influence on the firing rates of neurons with the potentiometric test, indicating high neuron-compatibility. It was then integrated with electrophysiology to simultaneously monitor DA variation and electrical signal in the brain, with stable monitoring of DA change in vivo for 8 weeks. The sensing fiber was flexible and stably worked after hundreds of bending, and it showed high sensitivity even after protein adsorption, thus offering a reliable tool for neuroscience.     

DNA-过氧化聚吡咯生物复合膜传感器的分析应用

在研究DNA与儿茶酚胺类分子之间相互作用的基础上, 以碳纤维电极(CFE)为基底, 制备了一种新型的DNA-过氧化聚吡咯(PPyox)生物复合膜传感器, 与单一的DNA或PPyox修饰层相比具有更高的灵敏度和选择性.     

Design and Evaluation of a Lactate Microbiosensor: Toward Multianalyte Monitoring of Neurometabolic Markers In Vivo in the Brain

Direct in vivo measurements of neurometabolic markers in the brain with high spatio-temporal resolution, sensitivity, and selectivity is highly important to understand neurometabolism. Electrochemical biosensors based on microelectrodes are very attractive analytical tools for continuous monitoring of neurometabolic markers, such as lactate and glucose in the brain extracellular space at resting and following neuronal activation. Here, we assess the merits of a platinized carbon fiber microelectrode (CFM/Pt) as a sensing platform for developing enzyme oxidase-based microbiosensors to measure extracellular lactate in the brain. Lactate oxidase was immobilized on the CFM/Pt surface by crosslinking with glutaraldehyde. The CFM/Pt-based lactate microbiosensor exhibited high sensitivity and selectivity, good operational stability, and low dependence on oxygen, temperature, and pH. An array consisting of a glucose and lactate microbiosensors, including a null sensor, was used for concurrent measurement of both neurometabolic substrates in vivo in the anesthetized rat brain. Rapid changes of lactate and glucose were observed in the cortex and hippocampus in response to local glucose and lactate application and upon insulin-induced fluctuations of systemic glucose. Overall, these results indicate that microbiosensors are a valuable tool to investigate neurometabolism and to better understand the role of major neurometabolic markers, such as lactate and glucose.     

Galvanic Redox Potentiometry for Fouling-Free and Stable Serotonin Sensing in a Living Animal Brain

Serotonin (5-HT) is a major neurotransmitter broadly involved in many aspects of feeling and behavior. Although its electro-activity makes it a promising candidate for electrochemical sensing, the persistent generation of fouling layers on the electrode by its oxidation products presents a hurdle for reliable sensing. Here, we present a fouling-free 5-HT sensor based on galvanic redox potentiometry. The sensor efficiently minimizes electrode fouling as revealed by in situ Raman spectroscopy, ensuring a less than 3 % signal change in a 2 hour continuous experiment, whereas amperometric sensors losing 90 % within 30 min. Most importantly, the sensor is highly amenable for in vivo studies, permitting real-time 5-HT monitoring, and supporting the mechanism associated with serotonin release in brain. Our system offers an effective way for sensing different neurochemicals having significant fouling issues, thus facilitating the molecular-level understanding of brain function.     

Recent advances in development of devices and probes for sensing and imaging in the brain

As the most important part of the central nervous system, the brain is extremely complex in structure and function. In vivo analysis of chemical signals is an essential way to investigate brain activity and function. Although functional magnetic resonance imaging(fMRI) or electrophysiology can be used to record brain activity, they are usually limited by low spatiotemporal fidelity or the difficulty of distinguishing the contributions of various neurochemicals. In addition, the development of in vivo biosensors with high selectivity and accuracy is essential to understand the roles that neurochemicals play in the brain. In this review, we focus on the development of instruments and devices for recording chemical signals in the live brain. Meanwhile,the strategies for development of electrochemical and fluorescent probes with high selectivity, high accuracy and good stability are also summarized. In particular, this review highlighted the contributions of our research group to this field. The development of techniques and probes enable us to understand the brain structure and function, and the mechanism of brain diseases, providing the solution for preventing and treating brain diseases.     

二茂铁-AQ修饰碳纤维微葡萄糖传感器的研究

本文用二茂铁、AQ成功地制备了微葡萄糖传感器。Eastman-AQ(AQ-55D,AQ29D)是一种新型的聚合物(磺酸酯)阳离于交换剂,涂于电极表面上,形成的膜除具有强的附着力外,同时还具有预富集、离子交换及防污性能。制得的电极具有制作方法简单、快速、重现性好,抗干扰能力强等特点。检测上限15.0 mmol/L,响应时间小于6s。由于AQ强的附着力,二茂铁及酶的流失较小。电极的稳定性有所提高。     

Facile and Large-scale Fabrication of Self-crimping Elastic Fibers for Large Strain Sensors

Stretchable conductive fibers offer unparalleled advantages in the development of wearable strain sensors for smart textiles due to their excellent flexibility and weaveability.However,the practical applications of these fibers in wearable devices are hindered by either contradictory properties of conductive fibers(high stretchability versus high sensing stability),or lack of manufacturing scalability.Herein,we present a facile approach for highly stretchable self-crimping fiber strain sensors based on a polyether-ester(TPEE)elastomer matrix using a side-by-side bicomponent melt-spinning process involving two parallel but attached components with different shrinkage properties.The TPEE component serves as a highly elastic mechanical support layer within the bicomponent fibers,while the conductive component(E-TPEE)of carbon black(CB),multiwalled carbon nanotubes(MWCNTs)and TPEE works as a strain-sensitive layer.In addition to the intrinsic elasticity of the matrix,the TPEE/E-TPEE bicomponent fibers present an excellent form of elasticity due to self-crimping.The self-crimping elongation of the fibers can provide a large deformation,and after the crimp disappears,the intrinsic elastic deformation is responsible for monitoring the strain sensing.The reliable strain sensing range of the TPEE/E-TPEE composite fibers was 160％-270％and could be regulated by adjusting the crimp structure.More importantly,the TPEE/E-TPEE fibers had a diameter of 30-40 μm and tenacity of 40-50 MPa,showing the necessary practicality.This work introduces new possibilities for fiber strain sensors produced in standard industrial spinning machines.     

A Novel Thin‐Film Copper Array Based on an Organic/Inorganic Sensor Hybrid: Microfabrication,Potentiometric Characterization,and Flow‐Injection Analysis Application

A first step towards the microfabrication of a thin‐film array based on an organic/inorganic sensor hybrid has been realized. The inorganic microsensor part incorporates a sensor membrane based on a chalcogenide glass material (Cu‐Ag‐As‐Se) prepared by pulsed laser deposition technique (PLD) combined with an PVC organic membrane‐based organic microsensor part that includes an o‐xylyene bis(N,N‐diisobutyl‐dithiocarbamate) ionophore. Both types of materials have been electrochemically evaluated as sensing materials for copper(II) ions. The integrated hybrid sensor array based on these sensing materials provides a linear Nernstian response covering the range 1×10^?6–1×10^?1 mol L^?1 of copper(II) ion concentration with a fast, reliable and reproducible response. The merit offered by the new type of thin‐film hybrid array includes the high selectivity feature of the organic membrane‐based thin‐film microsensor part in addition to the high stability of the inorganic thin‐film microsensor part. Moreover, the thin‐film sensor hybrid has been successfully applied in flow‐injection analysis (FIA) for the determination of copper(II) ions using a miniaturized home‐made flow‐through cell. Realization of the organic/inorganic thin‐film sensor hybrid array facilitates the development of a promising sophisticated electronic tongue for recognition and classification of various liquid media.     

Paracetamol voltammetric microsensors based on electrocopolymerized-molecularly imprinted film modified carbon fiber microelectrodes

A molecularly imprinted polymer is presented as a carbon fiber microelectrode coating for determining the presence of paracetamol. The polymeric film was obtained by electrocopolymerization of o-phenylenediamine and aniline in the presence of the template molecule, through the use of cyclic voltammetry. After removing the template, the signals of the microsensor were converted into physical ones by a voltammetric transductor using square wave voltammetry. Various parameters influencing the electropolymerization and voltammetric determination processes were examined and optimized. The response of the imprinted microsensor to paracetamol was linearly proportional to its concentration over the range 6.5 x 10(-6) to 2.0 x 10(-3) mol l(-1), with good stability and reproducibility (RSD < 5.6%). The detection limit was 1.5 microM. Under the experimental conditions used the voltammetric microsensor was able to differentiate between paracetamol and other closely structurally-related compounds present in biological fluids, such as certain catecholamines.     

Implantable optical fiber microelectrode with anti-biofouling ability for in vivo photoelectrochemical analysis

In-situ monitoring of neurochemicals is of vital importance for the understanding of brain functions. Microelectrode-based photoelectrochemical (PEC) sensing has emerged as a promising tool for in vivo analysis since it inherits the merits of both optical and electrochemical methods. However, the in-situ excitation of photoactive materials on the photoelectrode in living body is still a challenge because of limited tissue penetration depth of light. To circumvent this problem, we herein developed an implantable optical fiber (OF)-based microelectrode for in vivo PEC analysis. The working electrode was constructed by coating Au film as conducting layer and CdS@ZnO as photoactive material on a micron-sized OF, which was free of the limitation of light penetration in biological tissues. Further decoration of an anti-biofouling layer on the surface made the sensor robust in biosamples. It was successfully applied for monitoring Cu²⁺ level in three different brain regions in the rat model of cerebral ischemia/reperfusion.