Spatially resolved electrochemiluminescence on an array of electrode tips

An array of electrode tips with 6-microm center-to-center spacing, fabricated through chemical etching of an optical fiber bundle, and coated with gold, was used for initiating electrochemiluminescence (ECL) in an aqueous solution of Ru(bpy)3(2+) and tri-n-propylamine (TPrA). ECL generated at the tips of the electrodes in the array was detected with a CCD camera and exhibited both high sensitivity and high resolution. In the case in which the ECL signal could not be distinguished from the background, ECL signals could be obtained by pulsing the array and summing multiple CCD images. The behavior of this array was compared to a second array that consisted of individual electrodes insulated with an electrophoretic paint.     

Microring electrode/optical waveguide: electrochemical characterization and application to electrogenerated chemiluminescence

A novel optoelectrochemical microprobe has been developed that allows the combination of optical and electrochemical methods in a very small system. The key element is a gold-coated optical fiber polished to a flat surface, such that the gold forms a microring electrode surrounding the optical fiber. The electrochemistry of the microring has been characterized; cyclic voltammetric and chronoamperometric limiting currents agree well with published theoretical treatments. The probe has also been used for electrogenerated chemiluminescence with tris(2,2'-bypyridine)ruthenium(II) with persulfate in 50:50 MeCN:H2O. Detection limits of 4.3 X 10(-8) M Ru(bpy)3(2+) have been obtained.     

Fabrication and characterization of a nanometer-sized optical fiber electrode based on selective chemical etching for scanning electrochemical/optical microscopy

We have already reported a method for fabricating ultramicroelectrodes (Suzuki, K. JP Patent, 2004-45394, 2004). This method is based on the selective chemical etching of optical fibers. In this work, we undertake a detailed investigation involving a combination of etched optical fibers with various types of tapered tip (protruding-shape, double- (or pencil-) shape and triple-tapered electrode) and insulation with electrophoretic paint. Our goal is to establish a method for fabricating nanometer-sized optical fiber electrodes with high reproducibility. As a result, we realized pencil-shaped and triple-tapered electrodes that had radii in the nanometer range with high reproducibility. These nanometer-sized electrodes showed well-defined sigmoidal curves and stable diffusion-limited responses with cyclic voltammetry. The pencil-shaped optical fiber, which has a conical tip with a cone angle of 20 degrees , was effective for controlling the electrode radius. The pencil-shaped electrodes had higher reproducibility and smaller electrode radii (r(app) < 1.0 nm) than those of other etched optical fiber electrodes. By using a pencil-shaped electrode with a 105-nm radius as a probe, we obtained simultaneous electrochemical and optical images of an implantable interdigitated array electrode. We achieved nanometer-scale resolution with a combination of scanning electrochemical microscopy SECM and optical microscopy. The resolution of the electrochemical and optical images indicated sizes of 300 and 930 nm, respectively. The neurites of living PC12 cells were also successfully imaged on a 1.6-microm scale by using the negative feedback mode of an SECM.     

低损耗As-S玻璃光纤的制备与应用研究

采用反复蒸馏提纯技术和开放式动态蒸馏相结合的工艺, 制备了高纯As-S玻璃, 基本消除了玻璃在2.9、4和6.3 μm处的杂质吸收。利用旋转法制备出壁厚均匀、表面质量优异的硫系玻璃套管。采用棒管法拉制出丝径50 μm, 芯径40 μm具有芯包结构的硫系玻璃光纤。拉制的As-S光纤机械性能和光学性能优异, 光纤丝径波动小于1%, 弯曲半径优于4 mm, 中红外波段损耗基线小于0.5 dB/m。制备出像元呈正方形排列, 出端规格64×9, 入端规格192×3, 用于线-面转换的红外传像束, 像束断丝率为2.7%。利用该异型传像束成功实现了长线阵的红外推扫成像。     

Design and processing of high-density single-mode fiber arrays for imaging and parallel interferometer applications

The design and fabrication procedures for implementing a high-density (16-microm center spacing) single-mode fiber (SMF) array are described. The specific application for this array is a parallel optical coherence tomography system for endoscopic imaging. We obtained fiber elements by etching standard single-mode SMF-28 fibers to a diameter of 14-15 microm. We equalized 1-m lengths of fiber to within 1 mm by using a fiber interferometer setup, and we describe a method for packaging arrays with as many as 100 fibers.     

Optimization of the geometry and porosity of microelectrode arrays for sensor design

This paper describes the systematic investigation of a range of microelectrode arrays with varying dimensions fabricated by standard photolithographic and reactive-ion etching techniques. As expected from theory, the electrochemical behavior of microelectrode arrays with a constant individual diameter varied strongly with center-to-center spacing, the larger the spacing the more sigmoidal the recorded voltammogram. Furthermore, the behavior of arrays with a constant relative center-to-center spacing is shown to vary with individual electrode diameter, the arrays with the smallest electrodes producing strongly peaked voltammograms. Peak current densities and signal-to-noise ratios were also obtained for a variety of array geometries, and the use of electrodeposited platinum black electrodes was investigated. To demonstrate one advantage of using a loosely packed microelectrode array in electroanalysis, a ferrocene-mediated enzyme-linked assay involving the biocatalytic reduction of H2O2 was investigated. Results showed an improved temporal response, with current-time transients reaching a steady-state response more quickly using arrays with increased center-center spacings.     

前臂多腱肌的多通道表面肌电检测电极阵列装置的设计

设计了一种无创采集前臂多腱肌的多通道表面肌电（sEMG）信号的线性电极阵列．利用柔性印刷工艺制备的8个条形镀金电极以5mm的电极间距（IED）排成一列．预先冲孔的医用双面胶粘贴在电极面构成的一次性非接触粘贴结构可填充少量导电膏,而且便于电极固定在皮肤表面．单差分sEMG信号被放大和带通滤波．较小的电极阻抗保证了此种镀金电极具有恒定的电学性能．相对稳定的低电极．皮肤接触阻抗确保了稳定的基线和良好的信噪比（SNR）．为了探讨此装置的实际可用性,在单指力量输出任务时同步记录9名受试者指浅屈肌（FDS）的14通道sEMG信号．稳定的基线,良好的sEMG信号质量和显著的肌肉活动的通道差异性表明该电极阵列是一种可用于前臂多腱肌长时多通道sEMG信号采集的极具发展前景的技术．     

Micro-optical 1 x 4 fiber switch for multimode fibers with 600-microm core diameters

The design, manufacture, and test of a 1 x 4 micro-optical fiber switch for multimode fibers with 600-microm core diameters are described. Microlens array telescopes allow for variable and fast beam deflection when the positions of the cylindrical microlens arrays relative to each another are altered by specially designed piezomechanical actuators. Standard achromats are used for collimation of light emitted by the input multimode fiber and for focusing of the deflected light onto a linear array of output multimode fibers. Design and assembly of micro-optical as well as of optomechanical components are discussed. Insertion loss and cross talk are measured, and the results are compared with those of numerical optical simulations. Measurements of switching time and long-term stability, as well as of thermal behavior, are also presented.     

Soft microelectrode linear array for scanning electrochemical microscopy

A linear array of eight individual addressable microelectrodes has been developed in order to perform high-throughput scanning electrochemical microscopy (SECM) imaging of large sample areas in contact regime. Similar to previous reports, the soft microelectrode array was fabricated by ablating microchannels on a polyethylene terephthalate (PET) film and filling them with carbon ink. Improvements have been achieved by using a 5 μm thick Parylene coating that allows for smaller working distances, as the probe was mounted with the Parylene coating facing the sample surface. Additionally, the application of a SECM holder allows scanning in contact regime with a tilted probe, reducing the topographic effects and assuring the probe bending direction. The main advantage of the soft microelectrode array is the considerable decrease in the experimental time needed for imaging large sample areas. Additionally, soft microelectrode arrays are very stable and can be used several times, since the electrode surface can be regenerated by blade cutting. Cyclic voltammograms and approach curves were recorded in order to assess the electrochemical properties of the device. An SECM image of a gold on glass chip was obtained with high resolution and sensitivity, proving the feasibility of soft microelectrode arrays to detect localized surface activity. Finite element method (FEM) simulations were performed in order to establish the effect of diffusion layer overlapping between neighboring electrodes on the respective approach curves.     

Carbon-ring microelectrode arrays for electrochemical imaging of single cell exocytosis: fabrication and characterization

Fabrication of carbon microelectrode arrays, with up to 15 electrodes in total tips as small as 10-50 μm, is presented. The support structures of microelectrodes were obtained by pulling multiple quartz capillaries together to form hollow capillary arrays before carbon deposition. Carbon ring microelectrodes were deposited by pyrolysis of acetylene in the lumen of these quartz capillary arrays. Each carbon deposited array tip was filled with epoxy, followed by beveling of the tip of the array to form a deposited carbon-ring microelectrode array (CRMA). Both the number of the microelectrodes in the array and the tip size are independently tunable. These CRMAs have been characterized using scanning electron microscopy, energy dispersive X-ray spectroscopy, and electrogenerated chemiluminescence. Additionally, the electrochemical properties were investigated with steady-state voltammetry. In order to demonstrate the utility of these fabricated microelectrodes in neurochemistry, CRMAs containing eight microring electrodes were used for electrochemical monitoring of exocytotic events from single PC12 cells. Subcellular temporal heterogeneities in exocytosis (i.e. cold spots vs hot spots) were successfully detected with the CRMAs.     

Nanostructured gold microelectrodes for extracellular recording from electrogenic cells

We present a new biocompatible nanostructured microelectrode array for extracellular signal recording from electrogenic cells. Microfabrication techniques were combined with a template-assisted approach using nanoporous aluminum oxide to develop gold nanopillar electrodes. The nanopillars were approximately 300-400 nm high and had a diameter of 60 nm. Thus, they yielded a higher surface area of the electrodes resulting in a decreased impedance compared to planar electrodes. The interaction between the large-scale gold nanopillar arrays and cardiac muscle cells (HL-1) was investigated via focused ion beam milling. In the resulting cross-sections we observed a tight coupling between the HL-1 cells and the gold nanostructures. However, the cell membranes did not bend into the cleft between adjacent nanopillars due to the high pillar density. We performed extracellular potential recordings from HL-1 cells with the nanostructured microelectrode arrays. The maximal amplitudes recorded with the nanopillar electrodes were up to 100% higher than those recorded with planar gold electrodes. Increasing the aspect ratio of the gold nanopillars and changing the geometrical layout can further enhance the signal quality in the future.     

Microwell device for targeting single cells to electrochemical microelectrodes for high-throughput amperometric detection of quantal exocytosis

Electrochemical microelectrodes are commonly used to detect spikes of amperometric current that correspond to exocytosis of oxidizable transmitter from individual vesicles, i.e., quantal exocytosis. We are developing transparent multielectrochemical electrode arrays on microchips in order to automate measurement of quantal exocytosis. Here, we report development of an improved device to target individual cells to each microelectrode in an array. Efficient targeting (~75%) is achieved using cell-sized microwell traps fabricated in SU-8 photoresist together with patterning of poly(l-lysine) in register with electrodes to promote cell adhesion. The surface between electrodes is made resistant to cell adhesion using poly(ethylene glycol) in order to facilitate movement of cells to electrode "docking sites". We demonstrate the activity of the electrodes using the test analyte ferricyanide and perform recordings of quantal exocytosis from bovine adrenal chromaffin cells on the device. Multiple cell recordings on a single device demonstrate the consistency of spike measurements, and multiple recordings from the same electrodes demonstrate that the device can be cleaned and reused without degradation of performance. The new device will enable high-throughput studies of quantal exocytosis and may also find application in rapidly screening drugs or toxins for effects on exocytosis.     

An addressable microelectrode array for electrochemical detection

Column and row electrodes on two different glass substrates were orthogonally arranged in order to assemble an addressable microelectrode device for the purpose of comprehensive electrochemical detection. Amperometric signal at the individual crossing point of the column and row electrodes was detected separately on the basis of redox cycling of localized electroactive species occurring between the electrodes. The addressable microelectrode device was simple and could be easily assembled; however, it comprised as many as 10 x 10 addressable detection points on a single chip. The basic electrochemical performance of the device was investigated by using the ferricyanide/ferrocyanide redox couple. Electrochemical responses at 100 individual points could be collected within 22 s. The present device was successfully used for imaging the spots of alkaline phosphatase on the array substrate. The results indicate that the device can be applied to comprehensive and high-throughput detection and imaging of biochemical species.     

Development of an ordered array of optoelectrochemical individually readable sensors with submicrometer dimensions: application to remote electrochemiluminescence imaging

A novel array of optoelectrochemical submicrometer sensors for remote electrochemiluminescence (ECL) imaging is presented. This device was fabricated by chemical etching of a coherent optical fiber bundle to produce a nanotip array. The surface of the etched bundle was sputter-coated with a thin layer of indium tin oxide in order to create a transparent and electrically conductive surface that is insulated eventually by a new electrophoretic paint except for the apex of the tip. These fabrication steps produced an ordered array of optoelectrochemical sensors with submicrometer dimensions that retains the optical fiber bundle architecture. The electrochemical behavior of the sensor array was independently characterized by cyclic voltammetry and ECL experiments. The steady-state current indicates that the sensors are diffusively independent. This sensor array was further studied with a co-reactant ECL model system, such as Ru(bpy)(3)(2+)/TPrA. We clearly observed an ordered array of individual ECL micrometer spots, which corresponds to the sensor array structure. While the sensors of the array are not individually addressable electrochemically, we could establish that the sensors are optically independent and individually readable. Finally, we show that remote ECL imaging is performed quantitatively through the optoelectrochemical sensor array itself.     

Remote fluorescence imaging of dynamic concentration profiles with micrometer resolution using a coherent optical fiber bundle

Dynamic concentration profiles within the diffusion layer of an electrode were imaged in situ using fluorescence detection through a multichannel imaging fiber. In this work, a coherent optical fiber bundle is positioned orthogonal to the surface of an electrode and is used to report spatial and temporal micrometric changes in the fluorescence intensity of an initial fluorescent species. The fluorescence signal is directly related to the local concentration of a redox fluorescent reagent, which is electrochemically modulated by the electrode. Fluorescence images are collected through the optical fiber bundle during the oxidation of tris(2,2'-bipyridine)ruthenium(II) to ruthenium(III) at a diffusion-limited rate and allow the concentration profiles of Ru(II) reagent to be monitored in situ as a function of time. Tris(2,2'-bipyridine)ruthenium(II) is excited at 485 nm and emits fluorescence at 605 nm, whereas the Ru(III) oxidation state is not fluorescent. Our experiments emphasize the influence of two parameters on the micrometer spatial resolution: the numerical aperture of optical fibers within the bundle and the Ru(II) bulk concentration. The extent of the volume probed by each individual fiber of the bundle is discussed qualitatively in terms of a primary inner-filter effect and refractive index gradient. Experimentally measured fluorescence intensity profiles were found to be in very good agreement with concentration profiles predicted upon considering planar diffusion and thus validate the concept of this new application of imaging fibers. The originality of this remote approach is to provide a global view of the entire diffusion layer at a given time through one single image and to allow the time expansion of the diffusion layer to be followed quantitatively in real time.     

Identification of surface heterogeneity effects in cyclic voltammograms derived from analysis of an individually addressable gold array electrode

Voltammetric behavior at gold electrodes in aqueous media is known to be strongly dependent on electrode polishing and history. In this study, an electrode array consisting of 100 nominally identical and individually addressable gold disks electrodes, each with a radius of 127 microm, has been fabricated. The ability to analyze both individual electrode and total array performance enables microscopic aspects of the overall voltammetric response arising from variable levels of inhomogeneity in each electrode to be identified. The array configuration was initially employed with the reversible and hence relatively surface insensitive [Ru(NH 3) 6] (3+/2+) reaction and then with the more highly surface sensitive quasi-reversible [Fe(CN) 6] (3-/4-) process. In both these cases, the reactants and products are solution soluble and, at a scan rate of 50 mV s (-1), each electrode in the array is assumed to behave independently, since no evidence of overlapping of the diffusion layers was detected. As would be expected, the variability of the individual electrodes' responses was significantly larger than found for the summed electrode behavior. In the case of cytochrome c voltammetry at a 4,4'-dipyridyl disulfide modified electrode, a far greater dependence on electrode history and electrode heterogeneity was detected. In this case, voltammograms derived from individual electrodes in the gold array electrode exhibit shape variations ranging from peak to sigmoidal. However, again the total response was always found to be well-defined. This voltammetry is consistent with a microscopic model of heterogeneity where some parts of each chemically modified electrode surface are electroactive while other parts are less active. The findings are consistent with the common existence of electrode heterogeneity in cyclic voltammetric responses at gold electrodes, that are normally difficult to detect, but fundamentally important, as electrode nonuniformity can give rise to subtle forms of kinetic and other forms of dispersion.     

Enhanced two-photon excitation through optical fiber by single-mode propagation in a large core

Multiphoton excitation through optical fibers is limited by pulse broadening caused by self-phase modulation. We show that for short fiber lengths (approximately 2 m) two-photon excitation efficiency at the fiber output can be substantially improved by single-mode propagation in a large-area multimode fiber (10-microm core diameter) instead of a standard 5.5-microm core fiber. Measurements and numerical simulations of postfiber spectra and pulse widths demonstrate that the increase in efficiency is due to a reduction of nonlinear pulse broadening. Single-mode propagation in a large-core fiber is thus suitable for multiphoton applications for which pulse recompression is not possible at the fiber end.     

Fabrication and characterization of microwell array chemical sensors

Microwell arrays have been fabricated on the distal face of coherent fiber-optic bundles. A typical microwell array comprises approximately 3,000 individual optical fibers that were etched chemically. Individual microwells were 1 to 14-microm deep with approximately 22-microm widths and were filled partially with a chemical sensing (polymer + dye) layer to produce a microwell array sensor (MWAS). MWASs were fabricated using a technically expedient, photoinitiated polymerization reaction whereby a approximately 2 to 10-microm thick pH-sensitive or O2-sensitive sensing layer was immobilized inside each microwell. The pH-sensing layer comprised fluorescein isothiocyanate-dextran conjugate immobilized in a photopolymerizable poly(vinyl alcohol) membrane. The O2-sensing layer comprised a ruthenium metal complex entrapped in a gas-permeable photopolymerizable siloxane membrane. pH and PO2 were quantitated by acquiring luminescence images using an epifluorescence microscope/charge-coupled device imaging system. The pH-sensitive MWAS displayed a pKa of approximately 6.4 and a response time of approximately 2.5 s. The O2-sensitive MWAS behaved according to a nonlinear Stern-Volmer model with a maximum I0/I of approximately 4 and a response time of approximately 2.5 s. MWASs are advantageous in that suitably sized samples such as single biological cells can be co-localized with the sensing matrix in individual microwells.     

Experimental implementation of fiber optic bundle array wide FOV free space optical communications receiver

A gimbal-free wide field-of-regard (FOR) optical receiver has been built in a laboratory setting for proof-of-concept testing. Multiple datasets are presented that examine the overall FOR of the system and the receiver's ability to track and collect a signal from a moving source. The design is not intended to compete with traditional free space optical communication systems, but rather offer an alternative design that minimizes the number and complexity of mechanical components required at the surface of a small mobile platform. The receiver is composed of a micro-lens array and hexagonal bundles of large core optical fibers that route the optical signal to remote detectors and electronics. Each fiber in the bundle collects power from a distinct solid angle of space and a piezo-electric transducer is used to translate the micro-lens array and optimize coupling into a given fiber core in the bundle. The micro-lens to fiber bundle design is scalable, modular, and can be replicated in an array to increase aperture size.     

Highly sensitive detection of exocytotic dopamine release using a gold-nanoparticle-network microelectrode

Here we report a new type of microelectrode sensor for single-cell exocytotic dopamine release. The new microsensor is built by forming a gold-nanoparticle (AuNP) network on a carbon fiber microelectrode. First a gold surface is obtained on a carbon fiber microdisk electrode by partially etching away the carbon followed by electrochemical deposition of gold into the pore. The gold surface is chemically functionalized with a sol-gel silicate network derived from (3-mercaptopropyl)trimethoxysilane (MPTS). A AuNP network is formed by immobilizing Au nanoparticles onto the thiol groups in the sol-gel silicate network. The AuNP-network microelectrode has been characterized by scanning electron microscopy (SEM) and steady-state voltammetry. The AuNP-network microelectrode has been used for amperometric detection of exocytotic dopamine secretion from individual pheochromocytoma (PC12) cells. The results show significant differences in the kinetic peak parameters including shorter rise time, decay time, and half-width as compared to a bare carbon fiber electrode equivalent. These results indicate AuNP-network microelectrodes possess an excellent sensing activity for single-cell exocytotic catecholamine release, specifically dopamine. Moreover, key advantageous properties inherent to bare carbon fiber microelectrodes (i.e., rigidity, flexibility, and small size) are maintained in addition to an observed prolonged shelf life stability and resistance to cellular debris fouling and dopamine polymerization.