Nanotip Electrodes Research Articles

Nanoscale doping of polymeric semiconductors with confined electrochemical ion implantation.

Nanoresolved doping of polymeric semiconductors can overcome scaling limitations to create highly integrated flexible electronics, but remains a fundamental challenge due to isotropic diffusion of the dopants. Here we report a general methodology for achieving nanoscale ion-implantation-like electrochemical doping of polymeric semiconductors. This approach involves confining counterion electromigration within a glassy electrolyte composed of room-temperature ionic liquids and high-glass-transition-temperature insulating polymers. By precisely adjusting the electrolyte glass transition temperature (Tg) and the operating temperature (T), we create a highly localized electric field distribution and achieve anisotropic ion migration that is nearly vertical to the nanotip electrodes. The confined doping produces an excellent resolution of 56 nm with a lateral-extended doping length down to as little as 9.3 nm. We reveal a universal exponential dependence of the doping resolution on the temperature difference (Tg - T) that can be used to depict the doping resolution for almost infinite polymeric semiconductors. Moreover, we demonstrate its implications in a range of polymer electronic devices, including a 200% performance-enhanced organic transistor and a lateral p-n diode with seamless junction widths of <100 nm. Combined with a further demonstration in the scalability of the nanoscale doping, this concept may open up new opportunities for polymer-based nanoelectronics.

Application of a nanotip array-based electrochemical sensing platform for detection of indole derivatives as key indicators of gut microbiota health

The human gut microbiota plays a vital role in health and disease. Indole derivatives like indole, tryptamine and indoxyl sulfate represent important microbiota-associated metabolites and biomarkers of gut function. However, current techniques for analysis of these metabolites like mass spectrometry and HPLC suffer from limitations including high cost, complex sample preparation procedures, and long analysis times. In contrast, electrochemical sensing offers a rapid, simple and low-cost alternative with potential for point-of-care applications. We report a nanotip array-based electrochemical sensing platform for rapid, simple and sensitive quantification of indole derivatives. Silicon nanotip arrays were fabricated with controlled sub-micron morphology and coated with gold to form nanotip electrodes. Further modification with silver nanoparticles (AgNPs) led to significantly enhanced electrochemical signal. Using differential pulse voltammetry (DPV), distinct oxidation peaks were observed for indole, tryptamine and indoxyl sulfate, enabling sensitive detection down to nM levels. A broad linear dynamic range of 7 orders of magnitude demonstrated excellent quantitative capabilities. The sensor showed high reproducibility (<5 % RSD) and selectivity for the metabolites in the presence of potential interferents. Importantly, accurate detection of spiked indole derivatives was achieved in complex biological samples like serum and fecal extracts. The capacity to rapidly quantify indole derivatives in minimally processed samples highlights the potential of this nanotip electrochemical sensing strategy for point-of-care analysis of microbiota function. Overall, this work provides a robust nanotechnology-enabled platform for metabolite profiling to elucidate microbiota health and disease.

Simultaneous detection of vesicular content and exocytotic release with two electrodes in and at a single cell.

We developed a technique employing two electrodes to simultaneously and dynamically monitor vesicular neurotransmitter storage and vesicular transmitter release in and at the same cell. To do this, two electrochemical techniques, single-cell amperometry (SCA) and intracellular vesicle impact electrochemical cytometry (IVIEC), were applied using two nanotip electrodes. With one electrode being placed on top of a cell measuring exocytotic release and the other electrode being inserted into the cytoplasm measuring vesicular transmitter storage, upon chemical stimulation, exocytosis is triggered and the amount of release and storage can be quantified simultaneously and compared. By using this technique, we made direct comparison between exocytotic release and vesicular storage, and investigated the dynamic changes of vesicular transmitter content before, during, and after chemical stimulation of PC12 cells, a neuroendocrine cell line. While confirming that exocytosis is partial, we suggest that chemical stimulation either induces a replenishment of the releasable pool with a subpool of vesicles having higher amount of transmitter storage, or triggers the vesicles within the same subpool to load more transiently at approximately 10–20 s. Thus, a time scale for vesicle reloading is determined. The effect of l-3,4-dihydroxyphenylalanine (l-DOPA), the precursor to dopamine, on the dynamic alteration of vesicular storage upon chemical stimulation for exocytosis was also studied. We found that l-DOPA incubation reduces the observed changes of vesicular storage in regular PC12 cells, which might be due to an increased capacity of vesicular transmitter loading caused by l-DOPA. Our data provide another mechanism for plasticity after stimulation via quantitative and dynamic changes in the exocytotic machinery.

Comparison of Disk and Nanotip Electrodes for Measurement of Single-Cell Amperometry during Exocytotic Release.

We compare single-cell amperometric measurements of exocytosis from pheochromocytoma (PC12) cells between two types of electrodes, carbon fiber disk microelectrodes and nanotip conical-shape carbon fiber microelectrodes. During the exocytotic process, individual exocytotic release events, measured as current spikes at the electrode, offer quantitative and dynamic information about the chemical release from cells. Using two electrodes gives rise to an unequal distance between the fusion pore and the electrode as well as fusion pore size, which leads to different average spike shapes. Nanotip electrodes show a slightly higher and narrower spike than disk electrodes when measuring exocytosis. The estimated pore-electrode distance and fusion pore size for disk electrodes are 239 and 11.5 nm, while for nanotip electrodes, these are 215 and 18.2 nm, respectively. The data show that nanotip electrodes, despite showing slightly different dynamics for release, are quantitative in measuring the number of molecules released and can be used for quantitative comparison between exocytosis and vesicular content in intracellular vesicle impact electrochemical cytometry.

Intracellular injection of phospholipids directly alters exocytosis and the fraction of chemical release in chromaffin cells as measured by nano-electrochemistry.

Using a nano-injection method, we introduced phospholipids having different intrinsic geometries into single secretory cells and used single cell amperometry (SCA) and intracellular vesicle impact electrochemical cytometry (IVIEC) with nanotip electrodes to monitor the effects of intracellular incubation on the exocytosis process and vesicular storage. Combining tools, this work provides new information to understand the impact of intracellular membrane lipid engineering on exocytotic release, vesicular content and fraction of chemical release. We also assessed the effect of membrane lipid alteration on catecholamine storage of isolated vesicles by implementing another amperometric technique, vesicle impact electrochemical cytometry (VIEC), outside the cell. Exocytosis analysis reveals that the intracellular nano-injection of phosphatidylcholine and lysophosphatidylcholine decreases the number of released catecholamines, whereas phosphatidylethanolamine shows the opposite effect. These observations support the emerging hypothesis that lipid curvature results in membrane remodeling through secretory pathways, and also provide new evidence for a critical role of the lipid localization in modulating the release process. Interestingly, the IVIEC data imply that total vesicular content is also affected by in situ supplementation of the cells with some lipids, while, the corresponding VIEC results show that the neurotransmitter content in isolated vesicles is not affected by altering the vesicle membrane lipids. This suggests that the intervention of phospholipids inside the cell has its effect on the cellular machinery for vesicle release rather than vesicle structure, and leads to the somewhat surprising conclusion that modulating release has a direct effect on vesicle structure, which is likely due to the vesicles opening and closing again during exocytosis. These findings could lead to a novel regulatory mechanism for the exocytotic or synaptic strength based on lipid heterogeneity across the cell membrane.

Plasticity in exocytosis revealed through the effects of repetitive stimuli affect the content of nanometer vesicles and the fraction of transmitter released

Electrochemical techniques with disk and nano-tip electrodes, together with calcium imaging, were used to examine the effect of short-interval repetitive stimuli on both exocytosis and vesicular content in a model cell line. We show that the number of events decreases markedly with repeated stimuli suggesting a depletion of exocytosis machinery. However, repetitive stimuli induce a more stable fusion pore, leading to an increased amount of neurotransmitter release. In contrast, the total neurotransmitter content inside the vesicles decreases after repetitive stimuli, resulting in a higher average release fraction from each event. We suggest a possible mechanism regarding a link between activity-induced plasticity and fraction of release.

Combined Amperometry and Electrochemical Cytometry Reveal Differential Effects of Cocaine and Methylphenidate on Exocytosis and the Fraction of Chemical Release

AbstractAmperometry with nanotip electrodes has been applied to show cocaine and methylphenidate not only trigger declines in vesicle content and exocytotic catecholamine release in a model cell line but also differentially change the fraction of transmitter released from each individual vesicle. In addition, cocaine accelerates exocytotic release dynamics while they remain unchanged after methylphenidate treatment. The parameters from pre‐spike feet for the two drugs are also in opposition, suggesting this aspect of release is affected differentially. As cocaine and methylphenidate are psychostimulants with similar pharmacologic action but have opposite effects on cognition, these results might provide a missing link between the regulation of exocytosis and vesicles and the effect of this regulation on cognition, learning, and memory. A speculative chemical mechanism of the effect of these drugs on vesicle content and exocytosis is presented.

Combined Amperometry and Electrochemical Cytometry Reveal Differential Effects of Cocaine and Methylphenidate on Exocytosis and the Fraction of Chemical Release.

Amperometry with nanotip electrodes has been applied to show cocaine and methylphenidate not only trigger declines in vesicle content and exocytotic catecholamine release in a model cell line but also differentially change the fraction of transmitter released from each individual vesicle. In addition, cocaine accelerates exocytotic release dynamics while they remain unchanged after methylphenidate treatment. The parameters from pre‐spike feet for the two drugs are also in opposition, suggesting this aspect of release is affected differentially. As cocaine and methylphenidate are psychostimulants with similar pharmacologic action but have opposite effects on cognition, these results might provide a missing link between the regulation of exocytosis and vesicles and the effect of this regulation on cognition, learning, and memory. A speculative chemical mechanism of the effect of these drugs on vesicle content and exocytosis is presented.

Nanopore Opening at Flat and Nanotip Conical Electrodes during Vesicle Impact Electrochemical Cytometry.

The oxidation of catecholamine at a microelectrode, following its release from individual vesicles, allows interrogation of the content of single nanometer vesicles with vesicle impact electrochemical cytometry (VIEC). Previous to this development, there were no methods available to quantify the chemical load of single vesicles. However, accurate quantification of the content is hampered by uncertainty in the proportion of substituent molecules reaching the electrode surface (collection efficiency). In this work, we use quantitative modeling to calculate this collection efficiency. For all vesicles except those at the very edge of the electrode, modeling shows that ∼100% oxidation efficiency is achieved when employing a 33 μm diameter disk microelectrode for VIEC, independent of the location of the vesicle release pore. We use this to experimentally determine a precise distribution of catecholamine in individual vesicles extracted from PC12 cells. In contrast, we calculate that when a nanotip conical electrode (∼4 μm length, ∼1.5 μm diameter at the base) is employed, as in intracellular VIEC (IVIEC), the current-time response depends strongly on the position of the catecholamine-releasing pore in the vesicle membrane. When vesicle release occurs with the pore opening occurring far from the electrode, lower currents and partial oxidation (∼75%) of the catecholamine are predicted, as compared to higher currents and ∼100% oxidation, when the pore is close to/at the electrode surface. As close agreement is observed between the experimentally measured vesicular content in intracellular and extracted vesicles from the same cell line using nanotip and disk electrodes, respectively, we conclude that pores open at the electrode surface. Not only does this suggest that electroporation of the vesicle membrane is the primary driving force for catecholamine release from vesicles at polarized electrodes, but it also indicates that IVIEC with nanotip electrodes can directly assess vesicular content without correction.

Zinc Regulates Chemical-Transmitter Storage in Nanometer Vesicles and Exocytosis Dynamics as Measured by Amperometry.

We applied electrochemical techniques with nano-tip electrodes to show that micromolar concentrations of zinc not only trigger changes in the dynamics of exocytosis, but also vesicle content in a model cell line. The vesicle catecholamine content in PC12 cells is significantly decreased after 100 μm zinc treatment, but, catecholamine release during exocytosis remains nearly the same. This contrasts with the number of molecules stored in the exocytosis vesicles, which decreases, and we find that the amount of catecholamine released from zinc-treated cells reaches nearly 100 % content expelled. Further investigation shows that zinc slows down exocytotic release. Our results provide the missing link between zinc and the regulation of neurotransmitter release processes, which might be important in memory formation and storage.

Nano Secondary Ion Mass Spectrometry Imaging of Dopamine Distribution Across Nanometer Vesicles.

We report an approach to spatially resolve the content across nanometer neuroendocrine vesicles in nerve-like cells by correlating super high-resolution mass spectrometry imaging, NanoSIMS, with transmission electron microscopy (TEM). Furthermore, intracellular electrochemical cytometry at nanotip electrodes is used to count the number of molecules in individual vesicles to compare to imaged amounts in vesicles. Correlation between the NanoSIMS and TEM provides nanometer resolution of the inner structure of these organelles. Moreover, correlation with electrochemical methods provides a means to quantify and relate vesicle neurotransmitter content and release, which is used to explain the slow transfer of dopamine between vesicular compartments. These nanoanalytical tools reveal that dopamine loading/unloading between vesicular compartments, dense core and halo solution, is a kinetically limited process. The combination of NanoSIMS and TEM has been used to show the distribution profile of newly synthesized dopamine across individual vesicles. Our findings suggest that the vesicle inner morphology might regulate the neurotransmitter release event during open and closed exocytosis from dense core vesicles with hours of equilibrium needed to move significant amounts of catecholamine from the protein dense core despite its nanometer size.

Quantitative Chemical Measurements of Vesicular Transmitters with Electrochemical Cytometry.

Electrochemical cytometry adds a new dimension to our ability to study the chemistry and chemical storage of transmitter molecules stored in nanometer vesicles. The approach involves the adsorption and subsequent rupture of vesicles on an electrode surface during which the electroactive contents are quantitatively oxidized (or reduced). The measured current allows us to count the number of molecules in the vesicles using Faraday's law and to correlate this to the amount of molecules released when single exocytosis events take place at communicating cells. The original format for this method involved a capillary electrophoresis separation step to singly address each vesicle, but we have more recently discovered that cellular vesicles tend to adsorb to carbon electrodes and spontaneously as well as stochastically rupture to give mostly single vesicle events. This approach, called impact electrochemical cytometry, even though the impact is perhaps not the important part of this process, has been studied and the vesicle rupture appears to be at the interface between the vesicle and the electrode and is probably driven by electroporation. The pore size and rate of content electrolysis are a function of the pore diameter and the presence of a protein core in the vesicles. In model liposomes with no protein, events appear extremely rapidly as the soft nanoparticles impact the electrode and the contents are oxidized. It appears that the proteins decorating the surface of the vesicle are important in maintaining a gap from the electrode and when this gap is closed electroporation takes place. Models of the event response times suggest the pores formed are small enough so we can carry out these measurements at nanotip electrodes and we have used this to quantify the vesicle content in living cells in a mode we call intracellular impact electrochemical cytometry. The development of electrochemical cytometry allows comparison between vesicle content and vesicular release and we have found that only part of the vesicle content is released in typical exocytotic cases measured by amperometry. This has led to the novel hypothesis that most exocytosis from dense core vesicles is via mechanism where vesicles fuse with the cell membrane, some content is released and then close again to be reloaded and reused. It leaves open the possibility that cells regulate release during individual events. This might be important in learning and memory and be a nonreceptor pharmaceutical target for brain-related disorders. Indeed, the concept of the chemo-brain observed in cisplatin-treated cancer patients appears to be at least in part the result of changing the fraction of transmitter released and we have been able to show this by using the combined amperometric measurement of release and electrochemical cytometry at model cells.

Quantitative measurement of transmitters in individual vesicles in the cytoplasm of single cells with nanotip electrodes.

The quantification of vesicular transmitter content is important for studying the mechanisms of neurotransmission and malfunction in disease, and yet it is incredibly difficult to measure the tiny amounts of neurotransmitters in the attoliter volume of a single vesicle, especially in the cell environment. We introduce a novel method, intracellular vesicle electrochemical cytometry. A nanotip conical carbon-fiber microelectrode was used to electrochemically measure the total content of electroactive neurotransmitters in individual nanoscale vesicles in single PC12 cells as these vesicles lysed on the electrode inside the living cell. The results demonstrate that only a fraction of the quantal neurotransmitter content is released during exocytosis. These data support the intriguing hypothesis that the vesicle does not open all the way during the normal exocytosis process, thus resulting in incomplete expulsion of the vesicular contents.

Quantitative Measurement of Transmitters in Individual Vesicles in the Cytoplasm of Single Cells with Nanotip Electrodes

AbstractThe quantification of vesicular transmitter content is important for studying the mechanisms of neurotransmission and malfunction in disease, and yet it is incredibly difficult to measure the tiny amounts of neurotransmitters in the attoliter volume of a single vesicle, especially in the cell environment. We introduce a novel method, intracellular vesicle electrochemical cytometry. A nanotip conical carbon‐fiber microelectrode was used to electrochemically measure the total content of electroactive neurotransmitters in individual nanoscale vesicles in single PC12 cells as these vesicles lysed on the electrode inside the living cell. The results demonstrate that only a fraction of the quantal neurotransmitter content is released during exocytosis. These data support the intriguing hypothesis that the vesicle does not open all the way during the normal exocytosis process, thus resulting in incomplete expulsion of the vesicular contents.

Shearforce positioning of nanoprobe electrode arrays for scanning electrochemical microscopy experiments

Shearforce detection was applied to the controlled positioning of nanotip arrays at air/solid and liquid/solid interfaces in scanning electrochemical microscopy (SECM). The arrays were fabricated by wet chemical etching of ordered optical fiber bundles. The shearforce detection have been performed with a non-optical detection setup between 70 and 170kHz. The hydrodynamic nature of the shearforce interaction led to approach curves with length varying from few μm to more than 80μm, essentially controlled by the working frequency (resonance frequency), the nature of the medium (air, water, electrophoretic paint solution) and the surface state of the fiber bundle (cleaved or etched). This interface sensitive signal was applied to the positioning of the nanotip array in a 1μm PDMS film before the electrophoretic deposition of an insulating paint. The resulting nanoprobe electrode array, displaying nanotip electrodes individually insulated, has been characterized by cyclic voltammetry and SECM feedback curves. These results are discussed versus numerical simulations. They demonstrate that, except when in vicinity of a conductive substrate that may reveal the nanotip structure, the electrochemical behavior of the nanotip array is dictated by its micrometer dimension (the fiber bundle). In turn, they confirm the potential of shearforce detection for precise control over the array positioning.

Electrical properties of nanotip-assisted microplasma devices

Nanotip-assisted microplasma devices have been fabricated and characterized in Ne, Ar, or Ne/2% Ar. The electrical properties of the glow discharge in three different environments, Ne, Ar, and Ne/2% Ar, have been examined with direct current (dc) bipolar-pulsed waveforms with frequencies from 2to20kHz at pressures from 300to800torr. The operating voltage of the nanotip-assisted microplasma devices was decreased around 10–20V compared with devices without a nanotip electrode. The 6×1cm panel-type microplasma device with nanotip electrodes revealed low discharge voltages around 300V.

14-in Diagonal Size Nanotip-Enhanced Plasma Lamp in Liquid Crystal Display

The characteristics of nanotip-enhanced plasma lamps (NTL) as a backlight for liquid crystal displays have been investigated on a 14-in panel. Using a direct-current pulse bipolar waveform driving, the NTL operates with lower ignition and sustain voltages below 300 V compared with conventional plasma lamps due to the tiny discharge gap between the indium tin oxide electrode and nanotip electrodes. Luminous efficiencies of 16 lm/W at the lowest ignition voltage of 150 V have been realized for NTL, which correspond to 3780 of luminance in 260 V with 50 torr of pure xenon gas.