RESUMO
A multielectrode array (MEA) was fabricated with electrodes consisting of iridium oxide (IrOx) electrochemically deposited on nanoporous gold (NPG) to improve the moderate charge injection limit (ca. 1 mC cm-2) of NPG MEA. IrOx was electrodeposited by performing cyclic voltammetry with an IrOx deposition solution. The IrOx was electrodeposited on Au (EIROF/Au) and on NPG (EIROF/NPG) MEA, and the samples were analyzed in terms of the charge injection limit, charge storage capacity (CSC), and electrochemical impedance. The charge injection limit of the EIROF(100-cycled)/NPG MEA was estimated to be 2.3 mC cm-2 by measuring the voltage transient, and this value is sufficiently greater than the neural damage threshold (ca. 1 mC cm-2) and is also comparable to that of sputtered IrOx films. Considering the low charge injection limit (<0.1 mC cm-2) for the EIROF(100-cycled)/Au MEA, the high charge injection limit for the EIROF/NPG MEA was explained to be a result of synergetic combination of the inherently large surface area of the NPG and electrically active EIROF. The EIROF(100-cycled)/NPG exhibited an impedance of 9.7 ± 0.45 kΩ at 1 kHz and a CSC of 8 mC/cm-2, respectively, obtained via electrochemical impedance spectroscopy and integration of the cathodic current in a cyclic voltammogram. Scanning transmission electron microscopy and energy-dispersive X-ray spectroscopy are used to conduct an elemental mapping analysis of the cross-sectional structure of the EIROF/NPG and revealed that the EIROF had been uniformly deposited on the surface of the interconnected Au. The efficacy of the improvement in the charge injection limit of the EIROF/NPG MEA was evaluated with rat hippocampal slices. The EIROF/NPG electrodes exhibited a steeper increase in the negative peak amplitude of the field excitatory postsynaptic potentials (fEPSPs), even with an electrical stimulation of a lower amplitude (1-4 V), prolonged negative fEPSPs wave after peak response, and decreased serial reduction of fEPSPs compared to NPG MEA, all of which strongly indicate an improved charge injection for the EIROF/NPG MEA over NPG MEA.
RESUMO
In order to complement the high impedance electrical property of gold nanoparticles (Au NPs) we have performed electro-co-deposition of gold-platinum nanoparticles (Au-Pt NPs) onto the Au multi-electrode array (MEA) and modified the Au-Pt NPs surface with cell adhesive poly-D-lysine via thiol chemistry based covalent binding. The Au-Pt NPs were analyzed to have bimetallic nature not the mixture of Au NPs and Pt NPs by X-ray diffraction analysis and to have impedance value (4.0 × 10(4) Ω (at 1 kHz)) comparable to that of Pt NPs. The performance of Au-Pt NP-modified MEAs was also checked in relation to neuronal signal recording. The noise level in Au-Pt NP-modified MEAs was lower than in that of Au NP-modified MEA.
Assuntos
Técnicas Eletroquímicas , Ouro/química , Nanopartículas Metálicas/química , Neurônios/metabolismo , Platina/química , Transdução de Sinais , Animais , Células Cultivadas , Eletrodos , Neurônios/citologia , Ratos , Ratos Sprague-DawleyRESUMO
In this work, Cath.a-differentiated (CAD) cells were used in place of primary neuronal cells to assess the performance of vertically aligned carbon nanotubes (VACNTs) multi-electrode arrays (MEA). To fabricate high-performance MEA, VACNTs were directly grown on graphene/Pt electrodes via plasma enhanced chemical deposition technique. Here, graphene served as an intermediate layer lowering contact resistance between VACNTs and Pt electrode. In order to lower the electrode impedance and to enhance the cell adhesion, VACNTs-MEAs were treated with UV-ozone for 20 min. Impedance of VACNTs electrode at 1 kHz frequency exhibits a reasonable value (110 kΩ) for extracellular signal recording, and the signal to noise ratio the is good enough to measure low signal amplitude (15.7). Spontaneous firing events from CAD cells were successfully measured with VACNTs MEAs that were also found to be surprisingly robust toward the biological interactions.
Assuntos
Técnicas Citológicas/instrumentação , Eletrofisiologia/instrumentação , Nanotubos de Carbono/química , Animais , Células Cultivadas , Técnicas Citológicas/métodos , Eletrodos , Eletrofisiologia/métodos , Desenho de Equipamento , Grafite/química , CamundongosRESUMO
Herein, we fabricated fluorine-containing, polymer-based, flexible neural probes with fluorinated ethylene propylene (FEP) films as the substrates and photo-crosslinkable fluoropolymers as the passivation material. For fabrication, metal-free Au layer formation on the FEP film, the simultaneous photo-adhesion and photo-patterning technique, and the pulsed-laser scanning probe shaping technique were combined, followed by Au electrode surface modification. The resultant probes achieved a charge injection limit equal to 5.18 mC cm-2 by implementing iridium oxide-modified nanoporous Au (IrOx/NPG) structures. We performed simultaneous in vivo micro-stimulations of the Schaffer collateral fibres and recorded the evoked field excitatory postsynaptic potentials (fEPSPs) in the stratum radiatum layer of the hippocampal Cornu Ammonis 1 region using a single probe. Inducing the fEPSP at very low charge per pulse settings (3.2-3.6 nC/pulse) indicates the efficient charge injection capability of the IrOx/NPG electrode, thereby enabling safe, prolonged, and thrifty micro-stimulations. Furthermore, the single probe-induced and recorded long-term potentiation persisted for periods longer than 60 min following theta-burst stimulation. The materials used in this study are all biocompatible and chemically robust. The fabricated neural probes can be applied in chronic clinical trials in vivo.
Assuntos
Polímeros de Fluorcarboneto , Hipocampo , Hipocampo/fisiologia , Região CA1 Hipocampal , EletrodosRESUMO
We describe photopatterning technique that employs the photodegradation of cell-adhesive-modified poly(ethyleneimine) (m-PEI) to fabricate precise micropatterns on the indium tin oxide (ITO) substrate for guided neuronal growth. The photodegradation of m-PEI coated on hydroxyl group-terminated ITO substrate created micropatterns over a large area through deep UV irradiation. The photopatterned m-PEI layer can effectively guide neurite outgrowth and control neurite extensions from individual neurons.
Assuntos
Neurônios/citologia , Polietilenoimina/química , Adesão Celular , Processos Fotoquímicos , Compostos de Estanho/química , Raios UltravioletaRESUMO
We have developed a novel platform for selective binding of magnetic labels on planar Hall resistance sensor (PHR) for biosensing applications. The photoresist (PR) micro wells were prepared on the PHR sensor junctions to trap the magnetic bead at specified locations on the sensor surface and thin layer of Au was sputtered in the PR wells immobilize bimolecular. The Au surface is functionalized with single-stranded oligonucleotide and further biotin was used to immobilize streptavidin coated magnetic labels (Dynabeads Myone 1.0 microm, Invitrogen Co.). After removal of the PR wells on the sensor surface the non specific binding magnetic labels were successfully removed and only the chemically bounded magnetic labels were remained on the Au surface for detection of biomolecules using PHR sensor. We controlled the number of magnetic labels on the PHR sensor surface by using different sizes of the PR well on the junctions. The specifically bounded magnetic labels were successfully detected by characterizing the individual PHR sensor junctions. This technique enables the complete control over the magnetic labels for selective binding of biomolecules on the sensor surface for increasing the sensitivity of the PHR sensor as well as removal of the non specific bindings on the sensor surface.
RESUMO
The fluorescence emission properties of 2-(2'-hydroxy-4'-R-phenyl)benzothiazole (HBT-R) nanoparticles with different substituents (R = -COOH, -H, -CH(3), -OH, and -OCH(3)) were investigated using spectroscopic and theoretical methods. HBT-Rs displayed dual enol and keto (excited-state intramolecular proton transfer (ESIPT)) emissions in nonpolar solvents. The spectral change of their ESIPT emissions was affected differently by the electron donating (or withdrawing) power of the substituents; a bathochromic shift for the electron donating group and a hypsochromic shift in electron withdrawing group. In addition, the changes in energy levels calculated by the ab initio method were consistent with the spectral shifts of HBT-R in solution. We prepared aggregated HBT-R nanoparticles using a simple reprecipitation process in tetrahydrofuran-water solvents. The ESIPT emission of aggregated HBT-R nanoparticles was strongly enhanced (over 45 times) compared to those of monomer HBT-Rs in toluene, as markedly shifted ESIPT emissions are observed at longer wavelength without any quenching by self-absorption. Aggregated HBT-R nanoparticles showed longer lifetimes than those of monomer molecules. The temperature effect on the aqueous dispersion of the aggregated HBT-R nanoparticles was also explored. It shows a fluorescent ratiometric change in a range of temperature from 7 to 65 degrees C. A mechanism of a temperature-dependent equilibrium between the nanoparticles and the solvated enols is proposed for the emission color change.
RESUMO
Herein, we introduce a method to simultaneously photoadhere a photocrosslinkable polymer to a plasma-treated fluoropolymer while photopatterning the photocrosslinkable polymer via a single-photo-exposure as a new electrode passivation technique. Photoadhesion was determined to result from plasma-generated radicals of the plasma-treated fluoropolymer. Crystallinity of the fluoropolymer was analysed to determine the photoadhesion strength through its effects on both the formation of radicals and the etching of fluoropolymers. Passivation feasibility of simultaneous photoadhesion and photopatterning (P&P) technique were demonstrated by fabricating an Au electrocorticography electrode array and modifying the electrode with electro-deposited metallic nanoparticles. Adhesion of sputter-deposited Au to the fluoropolymer was dependent on mechanical interlocking, indicated by the formation of Au clusters which are typically influenced by the surface temperature during the sputter-deposition and the glass transition temperature of the fluoropolymer. The adhesion of Au to the fluoropolymer without an additional adhesion promotor and the proposed P&P passivation technique would help prevent detachment of the electrode and the delamination of the passivation layer in fluoropolymer-based neural electrode.
RESUMO
In order to improve the efficiency of infection of primary human endothelial cells in vitro of Kaposi's sarcoma-associated herpesvirus (KSHV), the effect of low speed centrifugation was investigated. The recombinant KSHV, BAC36, was used to examine the centrifugal enhancement of KSHV. Infectivity was estimated by green fluorescent protein (GFP) expression and real-time RT-PCR. The enhancement of infectivity was dependent upon the time and force of centrifugation in human umbilical vein endothelial cells (HUVECs). Centrifugation enhanced the infectivity of KSHV by up to 70 fold compared to non-centrifugal control infection for the same period of time; viral mRNA expression was also enhanced by centrifugation. HUVECs that were centrifuged before infection with KSHV displayed no enhancement in infectivity; therefore, enhancement is believed to occur during centrifugation. In addition, the mechanisms of infection including the initial viral attachment to cells, lipid rafts, and clathrin-mediated and caveolae endocytosis appear to be similar in KSHV infection with and without centrifugal enhancement. These results show that low speed centrifugation could be a useful tool for improving the efficiency of KSHV infection in vitro.
Assuntos
Centrifugação , Células Endoteliais/virologia , Herpesvirus Humano 8/crescimento & desenvolvimento , Virologia/métodos , Células Cultivadas , Fluorescência , Genes Reporter , Proteínas de Fluorescência Verde/metabolismo , Humanos , RNA Mensageiro/biossíntese , RNA Viral/biossíntese , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Ligação ViralRESUMO
Tetrodes, consisting of four twisted micro-wires can simultaneously record the number of neurons in the brain. To improve the quality of neuronal activity detection, the tetrode tips should be modified to increase the surface area and lower the impedance properties. In this study, tetrode tips were modified by the electrodeposition of Au nanoparticles (AuNPs) and dextran (Dex) doped poly (3,4-ethylenedioxythiophene) (PEDOT). The electrochemical properties were measured using electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV). A decrease in the impedance value from 4.3 MΩ to 13 kΩ at 1 kHz was achieved by the modified tetrodes. The cathodic charge storage capacity (CSCC) of AuNPs-PEDOT deposited tetrodes was 4.5 mC/cm2, as determined by CV measurements. The tetrodes that were electroplated with AuNPs and PEDOT exhibited an increased surface area, which reduced the tetrode impedance. In vivo recording in the ventral posterior medial (VPM) nucleus of the thalamus was performed to investigate the single-unit activity in normal rats. To evaluate the recording performance of modified tetrodes, spontaneous spike signals were recorded. The values of the L-ratio, isolation distance and signal-to-noise (SNR) confirmed that electroplating the tetrode surface with AuNPs and PEDOT improved the recording performance, and these parameters could be used to effectively quantify the spikes of each cluster.
RESUMO
A multichannel neural interface system is an important tool for various types of neuroscientific studies. For the electrical interface with a biological system, high-precision high-speed data recording and various types of stimulation capability are required. In addition, real-time signal processing is an important feature in the implementation of a real-time closed-loop system without unwanted substantial delay for feedback stimulation. Online spike sorting, the process of assigning neural spikes to an identified group of neurons or clusters, is a necessary step to make a closed-loop path in real time, but massive memory-space requirements commonly limit hardware implementations. Here, we present a 128-channel field-programmable gate array (FPGA)-based real-time closed-loop bidirectional neural interface system. The system supports 128 channels for simultaneous signal recording and eight selectable channels for stimulation. A modular 64-channel analog front-end (AFE) provides scalability and a parameterized specification of the AFE supports the recording of various electrophysiological signal types with 1.59 ± 0.76 root-mean-square noise. The stimulator supports both voltage-controlled and current-controlled arbitrarily shaped waveforms with the programmable amplitude and duration of pulse. An empirical algorithm for online real-time spike sorting is implemented in an FPGA. The spike-sorting is performed by template matching, and templates are created by an online real-time unsupervised learning process. A memory saving technique, called dynamic cache organizing, is proposed to reduce the memory requirement down to 6 kbit per channel and modular implementation improves the scalability for further extensions.
Assuntos
Próteses Neurais , Algoritmos , Animais , Córtex Cerebral/citologia , Sistemas Computacionais , Retroalimentação , Feminino , Humanos , Aprendizado de Máquina , Microeletrodos , Neurônios/fisiologia , Gravidez , Cultura Primária de Células , Ratos , Reprodutibilidade dos Testes , Processamento de Sinais Assistido por Computador , Razão Sinal-RuídoRESUMO
We covalently bound fluoropolymer (FP) films by plasma treatment followed by thermal pressing at temperatures below their melting point and fabricated an adhesion-metal-free flexible gold electrode array entirely encapsulated by the FP film, excepting the active electrode openings. The fabricated device was chemically resistant and was modified to have a lower impedance and efficient charge injection capability. The fabricated device was evaluated in vivo in rats and was confirmed to record the epidural epileptiform activity induced by chemical administration. The chemically inert nature of FPs and the gold electrode is expected to facilitate reliable neural interfacing without abiotic issues. Plasma treatment-induced covalent binding of FP films can also be utilized in a variety of applications requiring durability, such as implantable biosensors and sensor platforms operating under chemically harsh environments, including humid conditions.
RESUMO
Flexible supercapacitor electrodes have been fabricated by simple fabrication technique using graphite nanoflakes on polymer lapping films as flexible substrate. An additional thin layer of conducting polymer polypyrrole over the electrode improved the surface conductivity and exhibited excellent electrochemical performances. Such capacitor films showed better energy density and power density with a maximum capacitance value of 37 mF cm(-2) in a half cell configuration using 1 M H2SO4 electrolyte, 23 mF cm(-2) in full cell, and 6 mF cm(-2) as planar cell configuration using poly(vinyl alcohol) (PVA)/phosphoric acid (H3PO4) solid state electrolyte. Moreover, the graphite nanoflakes/polypyrrole over polymer lapping film demonstrated good flexibility and cyclic stability.
RESUMO
OBJECTIVE: Nanoporous gold (Au) structures can reduce the impedance and enhance the charge injection capability of multi-electrode arrays (MEAs) used for interfacing neuronal networks. Even though there are various nanoporous Au preparation techniques, fabrication of MEA based on low-cost electro-codeposition of Ag:Au has not been performed. In this work, we have modified a Au MEA via the electro-codeposition of Ag:Au alloy, followed by the chemical etching of Ag, and report on the in vitro extracellular recording and stimulation performance of the nanoporous Au-modified MEA. APPROACH: Ag:Au alloy was electro-codeposited on a bilayer lift-off resist sputter-deposition passivated Au MEA followed by chemical etching of Ag to form a porous Au structure. MAIN RESULTS: The porous Au structure was analyzed by scanning electron microscopy and tunneling electron microscopy and found to have an interconnected nanoporous Au structure. The impedance value of the nanoporous Au-modified MEA is 15.4 ± 0.55 kΩ at 1 kHz, accompanied by the base noise V rms of 2.4 ± 0.3 µV. The charge injection limit of the nanoporous Au-modified electrode estimated from voltage transient measurement is approximately 1 mC cm(-2), which is comparable to roughened platinum and carbon nanotube electrodes. The charge injection capability of the nanoporous Au-modified MEA was confirmed by observing stimulus-induced spikes at above 0.2 V. The nanoporous Au-modified MEA showed mechanical durability upon ultrasonic treatment for up to an hour. SIGNIFICANCE: Electro-codeposition of Ag:Au alloy combined with chemical etching Ag is a low-cost process for fabricating nanoporous Au-modified MEA suitable for establishing the stimulus-response relationship of cultured neuronal networks.
Assuntos
Líquido Extracelular/fisiologia , Ouro/química , Nanopartículas Metálicas/química , Neurônios/fisiologia , Animais , Células Cultivadas , Estimulação Elétrica/métodos , Microeletrodos , Porosidade , Ratos , Ratos Sprague-DawleyRESUMO
The photoreduction of water-soluble cationic manganese(III) meso-tetrakis(1-methylpyridium-4-yl)porphyrin (Mn(III)(TMPyP)(4+)) bound to a synthetic polynucleotide, either poly[d(A-T)2] or poly[d(G-C)2], was examined by conventional absorption and circular dichroism (CD) spectroscopy, transient absorption, and transient Raman spectroscopy. Upon binding, Mn(III)(TMPyP)(4+) produced a positive CD signal for both polynucleotides, suggesting external binding. In the poly[d(A-T)2]-Mn(III)(TMPyP)(4+) adduct case, an interaction between the bound porphyrin was suggested. The transient absorption spectral features of Mn(III)(TMPyP)(4+) in the presence of poly[d(A-T)2] and poly[d(G-C)2] were similar to those of the photoreduced products, Mn(II)(TMPyP)(4+), whereas Mn(III)(TMPyP)(4+) in the absence of polynucleotides retained its oxidation state. This indicated that both poly[d(A-T)2] and poly[d(G-C)2] act as electron donors, resulting in photo-oxidized G and A bases. The transient Raman bands (ν2 and ν4) that were assigned to porphyrin macrocycles exhibited a large downshift of ~25 cm(-1), indicating the photoreduction of Mn(III) to Mn(II) porphyrins when bound to both polynucleotides. The transient Raman bands for pyridine were enhanced significantly, suggesting that the rotation of peripheral groups for binding with polynucleotides is the major change in the geometry expected in the photoreduction process. These photoinduced changes do not appear to be affected by the binding mode of porphyrin.
Assuntos
Complexos de Coordenação/química , Manganês/química , Polidesoxirribonucleotídeos/química , Porfirinas/química , Pareamento de Bases , Dicroísmo Circular , Complexos de Coordenação/síntese química , Transporte de Elétrons , Luz , Oxirredução , Polidesoxirribonucleotídeos/síntese química , Análise Espectral Raman , TermodinâmicaRESUMO
We have synthesized photocrosslinkable benzophenone copolymer, Poly(St-co-MBz), and fabricated cell-repellent patterns of Poly(St-co-MBz) on covalently bound poly-D-lysine (PDL) layer via the photocrosslinking. We have successfully obtained fine grid line pattern with line width of 3 µm and fine neurite, presumably axon, patterns with excellent pattern fidelity. We found that benzophenone unit can be crosslinked under the exposure of UV (with the intensity of â¼77 mW/cm² at 280 nm and â¼60 mW/cm² at 365 nm) without photo-oxidative damage to PDL, poly-L-lysine, and polyethyleneimine.
Assuntos
Movimento Celular/fisiologia , Neuritos/fisiologia , Neurônios/citologia , Polilisina/química , Polímeros/química , Animais , Benzofenonas/farmacologia , Células Cultivadas , Córtex Cerebral/citologia , Embrião de Mamíferos , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Fármacos Fotossensibilizantes , Ratos , Ratos Sprague-Dawley , Espectrometria de Fluorescência , Tubulina (Proteína)/metabolismo , Raios UltravioletaRESUMO
We have prepared the poly-d-lysine (PDL) bound surfaces for neuron cell culture by covalent binding between the poly-d-Lysine and substrates and investigated neuronal cell adhesion properties and cell growth morphology. The number of neuronal cell and the number of neurite per neuronal cell on PDL bound surfaces was much more than those on PDL coated surfaces and also the neuronal cells on PDL bounded surfaces survived a longer time. On the pattern of covalently bound PDL, neuronal cells and their neurites are confined within the grid line leading to patterned neuronal networks with the long-term survival.
Assuntos
Adesão Celular/fisiologia , Técnicas de Cultura de Células/métodos , Neurônios/citologia , Polilisina/química , Animais , Proliferação de Células , Sobrevivência Celular , Células Cultivadas , Imunofluorescência , Índio/química , Ratos , Ratos Sprague-Dawley , Dióxido de Silício/química , Compostos de Estanho/químicaRESUMO
A new class of organic nanoparticles (CN-MBE nanoparticles) with a mean diameter of ca. 30-40 nm, which exhibit a strongly enhanced fluorescence emission, were prepared by a simple reprecipitation method. CN-MBE (1-cyano-trans-1,2-bis-(4'-methylbiphenyl)ethylene) is very weakly fluorescent in solution, but the intensity is increased by almost 700 times in the nanoparticles. Enhanced emission in CN-MBE nanoparticles is attributed to the synergetic effect of intramolecular planarization and J-type aggregate formation (restricted excimer formation) in nanopaticles. On/off fluorescence switching for organic vapor was demonstrated with CN-MBE nanoparticles.