Formation of a nanometer-wide gap between tip and substrate electrodes by scanning electrochemical microscopy (SECM) enables voltammetric measurement of ultrafast electron-transfer kinetics. Herein, we demonstrate the advantage of SECM-based nanogap voltammetry to assess the cleanness of the substrate surface in solution by confirming that airborne contamination of highly oriented pyrolytic graphite (HOPG) causes the nonideal asymmetry of paired nanogap voltammograms of (ferrocenylmethyl)trimethylammonium (Fc(+)). We hypothesize that the amperometric response of a 1 μm-diameter Pt tip is less enhanced in the feedback mode, where more hydrophilic Fc(2+) is generated from Fc(+) at the tip and reduced voltammetrically at the HOPG surface covered with airborne hydrophobic contaminants. The tip current is more enhanced in the substrate generation/tip collection mode, where less charged Fc(+) is oxidized at the contaminated HOPG surface. In fact, symmetric pairs of nanogap voltammograms are obtained with the cleaner HOPG surface that is exfoliated in humidified air and covered with a nanometer-thick water adlayer to suppress airborne contamination. This result disproves a misconception that the asymmetry of paired nanogap voltammograms is due to electron exchange mediated by Fc(2+) adsorbed on the glass sheath of the tip. Moreover, weak Fc(+) adsorption on the HOPG surface causes only the small hysteresis of each voltammogram upon forward and reverse sweeps of the HOPG potential. Significantly, no Fc(2+) adsorption on the HOPG surface ensures that the simple outer-sphere pathway mediates ultrafast electron transfer of the Fc(2+/+) couple with standard rate constants of ≥12 cm/s as estimated from symmetric pairs of reversible nanogap voltammograms.

Electron-beam (e-beam) deposition of carbon on a gold substrate yields a very flat (0.43 nm of root-mean-square roughness), amorphous carbon film consisting of a mixture of sp- and sp-hybridized carbon with sufficient conductivity to avoid ohmic potential error. E-beam carbon (eC) has attractive properties for conventional electrochemistry, including low background current and sufficient transparency for optical spectroscopy. A layer of KCl deposited by e-beam to the eC surface without breaking vacuum protects the surface from the environment after fabrication until dissolved by an ultrapure electrolyte solution. Nanogap voltammetry using scanning electrochemical microscopy (SECM) permits measurement of heterogeneous standard electron-transfer rate constants (k°) in a clean environment without exposure of the electrode surface to ambient air. The ultraflat eC surface permitted nanogap voltammetry with very thin electrode-to-substrate gaps, thus increasing the diffusion limit for k° measurement to >14 cm/s for a gap of 44 nm. Ferrocene trimethylammonium as the redox mediator exhibited a diffusion-limited k° for the previously KCl-protected eC surface, while k° was 1.45 cm/s for unprotected eC. The k° for Ru(NH) increased from 1.7 cm/s for unprotected eC to above the measurable limit of 6.9 cm/s for a KCl-protected eC electrode.

Nanoscale scanning electrochemical microscopy (SECM) is a powerful scanning probe technique that enables high-resolution imaging of chemical processes at single nanometer-sized objects. However, it has been a challenging task to quantitatively understand nanoscale SECM images, which requires to accurately characterize the size and geometry of nanoelectrode tips. Herein, we address this challenge through transmission electron microscopy (TEM) of quartz nanopipets for SECM imaging of single solid-state nanopores by using nanopipet-supported interfaces between two immiscible electrolyte solutions (ITIES) as tips. We take advantage of the high resolution of TEM to demonstrate that laser-pulled quartz nanopipets reproducibly yield not only an extremely small tip diameter of ~30 nm, but also a substantial tip roughness of ~5 nm. The size and roughness of a nanopipet can be reliably determined by optimizing the intensity of the electron beam not to melt or deform the quartz nanotip without metal coating. Electrochemically, the nanoscale ITIES supported by a rough nanotip gives higher amperometric responses to tetrabutylammonium than expected for a 30 nm-diameter disk tip. The finite element simulation of sphere-cap ITIES tips accounts for the high current responses and also reveals that the SECM images of 100 nm-diameter Si3N4 nanopores are enlarged along the direction of the tip scan. Nevertheless, spatial resolution is not significantly compromised by a sphere-cap tip, which can be scanned in closer proximity to the substrate. This finding augments the utility of a protruded tip, which can be fabricated and miniaturized more readily to facilitate nanoscale SECM imaging.

The nuclear pore complex (NPC) solely mediates molecular transport between the nucleus and cytoplasm of a eukaryotic cell to play important biological and biomedical roles. It, however, is not well understood chemically how this biological nanopore selectively and efficiently transports various substances including small molecules, proteins, and RNAs by using transport barriers that are rich in highly disordered repeats of hydrophobic phenylalanine and glycine intermingled with charged amino acids. Herein, we employ scanning electrochemical microscopy to image and measure the high permeability of NPCs to small redox molecules. The effective medium theory demonstrates that the measured permeability is controlled by diffusional translocation of probe molecules through water-filled nanopores without steric or electrostatic hindrance from hydrophobic or charged regions of transport barriers, respectively. The permeability of NPCs, however, is lowered by a low millimolar concentration of Ca 2+ , which can interact with anionic regions of transport barriers to alter their spatial distributions within the nanopore. We employ atomic force microscopy to confirm that transport barriers of NPCs are dominantly recessed (~80%) or entangled (~20%) at the high Ca 2+ level in contrast to authentic populations of entangled (~50%), recessed (~25%), and "plugged" (~25%) conformations at a physiological Ca 2+ level of sub-micromolar. We propose a model for synchronized Ca 2+ effects on the conformation and permeability of NPCs, where transport barriers are viscosified to lower permeability. Significantly, this result supports a hypothesis that the functional structure of transport barriers is maintained not only by their hydrophobic regions, but also by charged regions.

Applications of conducting carbon materials for highly efficient electrochemical energy devices require a greater fundamental understanding of heterogeneous electron-transfer (ET) mechanisms. This task, however, is highly challenging experimentally, because an adsorbing carbon surface may easily conceal its intrinsic reactivity through adventitious contamination. Herein, we employ nanoscale scanning electrochemical microscopy (SECM) and cyclic voltammetry to gain new insights into the interplay between heterogeneous ET and adsorption of a Co(III)/Co(II)-complex redox couple at the contamination-free surface of electron-beam-deposited carbon (eC). Specifically, we investigate the redox couple of tris(1,10-phenanthroline)cobalt(II), Co(phen), as a promising mediator for dye-sensitized solar cells and redox flow batteries. A pristine eC surface overlaid with KCl is prepared in vacuum, protected from contamination in air, and exposed to an ultrapure aqueous solution of Co(phen) by the dissolution of the protective KCl layer. We employ SECM-based nanogap voltammetry to quantitatively demonstrate that Co(phen) is adsorbed on the pristine eC surface to electrostatically self-inhibit outer-sphere ET of nonadsorbed Co(phen) and Co(phen). Strong electrostatic repulsion among Co(phen) adsorbates is also demonstrated by SECM-based nanogap voltammetry and cyclic voltammetry. Quantitatively, self-inhibitory ET is characterized by a linear decrease in the standard rate constant of Co(phen) oxidation with a higher surface concentration of Co(phen) at the formal potential. This unique relationship is consistent not with the Frumkin model of double layer effects, but with the Amatore model of partially blocked electrodes as extended for self-inhibitory ET. Significantly, the complicated coupling of electron transfer and surface adsorption is resolved by combining nanoscale and macroscale voltammetric methods.

Electrochemical measurements with unprecedentedly high sensitivity, selectivity, and kinetic resolution have been enabled by a pair of electrodes separated by a nanometer-wide gap. The fabrication of nanogap electrodes, however, requires extensive nanolithography or nanoscale electrode positioning, thereby preventing the full exploration of this powerful method in electrode design and application. Herein, we report the simple fabrication of double-carbon-fiber ultramicroelectrodes (UMEs) with nanometer-wide gaps not only to facilitate nanogap-based electrochemical measurements, but also to gain high time resolution, signal-to-background ratio, and kinetic selectivity for dopamine against ascorbic acid. Specifically, ~7 μm-diameter carbon fibers are inserted into a double-bore glass capillary, heat-pulled, and milled by focused-ion-beam technology to yield ~50 μm-long double-cylinder UMEs. The redox cycling of the Ru(NH3)63+/2+ couple across a nanogap between voltammetric generator and amperometric collector electrodes reaches quasi-steady states at fast scan rates of 100 V/s as demonstrated experimentally and even 1000 V/s as predicted theoretically. The transient background of the amperometric collector response is suppressed ~100 times in comparison with that of the voltammetric generator response. Nanogap voltammograms based on the collector response against the cycled generator potential are quantitatively analyzed without background subtraction to reproducibly yield nanogap widths of ~0.18 μm and a standard electron-transfer rate constant of 0.9 cm/s. Moreover, nanogap-mediated redox cycling can be initiated by dopamine oxidation at the generator electrode to largely improve the dopamine selectivity of the collector response against ascorbic acid, which is also oxidized at the generator electrode to immediately and irreversibly produce a redox-inactive species.

High temporal resolution of fast-scan cyclic voltammetry (FSCV) is widely appreciated in fundamental and applied electrochemistry to quantitatively investigate rapid dynamics of electron transfer and neurotransmission using ultramicroelectrodes (UMEs). Faster potential scan, however, linearly increases the background current, which must be subtracted for quantitative FSCV. Herein, we numerically simulate fast-scan nanogap voltammetry (FSNV) for quantitative detection of diffusing redox species under quasi-steady states without the need of background subtraction while maintaining high temporal resolution of transient FSCV. These advantages of FSNV originate from the use of a parallel pair of cylindrical UMEs with nanometer-wide separation in contrast to FSCV with single UMEs. In FSNV, diffusional redox cycling across the nanogap is driven voltammetrically at the generator electrode and monitored amperometrically at the collector electrode without the transient background. We reveal that the cylindrical collector electrode can reach quasi-steady states ~104 times faster than the generator electrode with identical sizes to allow for fast scan. Double-microcylinder and nanocylinder UMEs enable quasi-steady-state FSNV at hundreds volts per second as practiced for in-vivo FSCV and megavolts per second as achieved for ultra-FSCV, respectively. Rational design and simple fabrication of double-cylinder UMEs are proposed to broaden the application of nanogap voltammetry.

Scanning electrochemical microscopy (SECM) enables high-resolution imaging by examining the amperometric response of an ultramicroelectrode tip near a substrate. Spatial resolution, however, is compromised for non-flat substrates, where distances from a tip far exceed the tip size to avoid artifacts caused by the tip-substrate contact. Herein, we propose a new imaging mode of SECM based on real-time analysis of approach curve to actively control nanoscale tip-substrate distances without contact. The power of this software-based method is demonstrated by imaging an insulating substrate with step edges using standard instrumentation without combination of another method for distance measurement, e.g., atomic force microscopy. An ~500 nm-diameter Pt tip approaches down to ~50 nm from upper and lower terraces of a 500 nm-height step edge, which are located by real-time theoretical fitting of experimental approach curve to ensure the lack of electrochemical reactivity. The tip approach to step edge can be terminated at <20 nm prior to the tip-substrate contact as soon as the theory deviates from the tip current, which is analyzed numerically afterward to locate the inert edge. The advantageous local adjustment of tip height and tip current at the final point of tip approach distinguishes the proposed imaging mode from other modes based on standard instrumentation. In addition, the glass sheath of Pt tip is thinned to ~150 nm to rarely contact the step edge, which is unavoidable and instantaneously detected as an abrupt change in the slope of approach curve to prevent the damage of fragile nanotip.