We report on real-time electrochemical detection of individual DNA hybridization events at an electrode surface. The experiment is carried out in a microelectrochemical device configured with a working electrode modified with single-stranded DNA probe molecules. When a complementary DNA strand labelled with a catalyst hybridizes to the probe, an easily detectable electrocatalytic current is observed. In the experiments reported here, the catalyst is a platinum nanoparticle and the current arises from electrocatalytic oxidation of hydrazine. Two types of current transients are observed: short bursts and longer-lived steps. At low concentrations of hydrazine, the average size of the current transients is proportional to the amount of hydrazine present, but at higher concentrations the hydrazine oxidation reaction interferes with hybridization.

No abstract

We report on the effect of convection on electrochemically active collisions between individual Pt nanoparticles (PtNPs) and Hg and Au electrodes. Compared to standard electrochemical cells utilizing Hg and Au ultramicroelectrodes (UMEs) used in previous studies of electrocatalytic amplification, microelectrochemical devices offer two major advantages. First, the PtNP limit of detection (0.084 pM) is ∼8 times lower than the lowest concentration measured using UMEs. Second, convection enhances the mass transfer of PtNPs to the electrode surface, which enhances the collision frequency from ∼0.02 pM(-1) s(-1) on UMEs to ∼0.07 pM(-1) s(-1) in microelectrochemical devices. We also show that the size of PtNPs can be measured in flowing systems using data from collision experiments and then validate this finding using multiphysics simulations.

We report electrochemical catalytic amplification of individual collisions between ∼57 nm diameter Pt nanoparticles (Pt NPs) and 12.5 μm diameter Au ultramicroelectrodes modified with passivating, electrostatically assembled polyelectrolyte multilayer (PEM) films prepared by the layer-by-layer deposition method. Two key findings are reported. First, despite the thicknesses of the insulating PEM films, which range up to 5 nm, electrons are able to tunnel from the Pt NPs to the electrode resulting in electrocatalytic N2H4 oxidation at the PEM film-solution interface. These single-particle measurements are in accord with prior reports showing that the electrochemical activity of passive PEM films can be reactivated by adsorption of metallic NPs. Second, it is possible to control the frequency of the collisions by manipulating the net electrostatic charge present on the outer surface of the PEM thin film. These results, which demonstrate that chemistry can be used to control electrocatalytic amplification, set the stage for future sensing applications.

We report electrochemical detection of collisions between individual magnetic microbeads, present at subattomolar concentrations, and electrode surfaces. This limit of detection is 4 orders of magnitude lower than has been reported previously, and it is enabled by using a magnetic field to preconcentrate the microbeads prior to detection in a microfluidic electrochemical cell. Importantly, the frequency of collisions between the microbeads and the electrode is not compromised by the low concentration of microbeads. These findings represent an unusual case of detecting individual electrochemical events at very low analyte concentration. In addition to experiments supporting these claims, finite-element simulations provide additional insights into the nature of the interactions between flowing microbeads and their influence on electrochemical processes.

An example of proton-coupled electron transfer (PCET) comprised by the electrochemical oxidation of 1,4-hydroquinone (1,4-H 2 Q) in acetonitrile was studied in the presence of Brønsted bases in acetonitrile. Of the two types of bases studied, the negatively charged carboxylates, trifluoroacetate (TFAC À ), benzoate (BZ À ), and acetate (AC À ), showed hydrogen bonding with 1,4-H 2 Q, whereas the neutral amines, pyridine (PY) and N, N 0 -diisopropylethylamine (DIPEA), did not. This difference allowed a unique investigation of the effect of proton transfer on PCET with and without the influence of hydrogen bonding using two bases (TFAC À and PY) with approximately the same pK a (∼12). The study revealed that hydrogen bonding of 1,4-H 2 Q with the base TFAC À made the half wave redox potential of 1,4-H 2 Q more negative (easier to oxidize) by 0.186 V with respect to the oxidation in the presence of the same concentration of added PY, which does not hydrogen bond with 1,4-H 2 Q. Both types of bases studied, carboxylates and amines, showed a combination of kinetic and thermodynamic effects in the oxidation voltammerty of 1,4-H 2 Q; however, no evidence of concerted pathways was found at the conditions studied as indicated by H/D kinetic isotope experiments. The mechanism from fitted digital simulations for all the bases supports a stepwise PCET, even in the presence of hydrogen bonding, implying that the latter does not prevail in the transition state nor is rate determining. Hydrogen bonding was verified by 1 H NMR spectroscopy, while the electrochemical studies were carried out by cyclic voltammetry. The hydrogen bonding constants and diffusion coefficients determined by 1 H NMR were used in digital simulations that were fitted to experimental voltammograms.

Here we report on the effect of DNA modification on individual collisions between Pt nanoparticles (PtNPs) and ultramicroelectrode (UME) surfaces. These results extend recent reports of electrocatalytic amplification (ECA) arising from collisions between naked surfaces, and they are motivated by our interest in using ECA for low-level biosensing applications. In the present case, we studied collisions between naked PtNPs and DNA-modified Au and Hg UMEs and also collisions between DNA-modified PtNPs and naked Au and Hg UMEs. In all cases, the sensing reaction is the catalytic oxidation of N2H4. The presence of ssDNA (5-mer or 25-mer) immobilized on the UME surface has little effect on the magnitude or frequency of ECA signals, regardless of whether the electrode is Au or Hg. In contrast, when DNA is immobilized on the PtNPs and the electrodes are naked, clear trends emerge. Specifically, as the surface concentration of ssDNA on the PtNP surface increases, the magnitude and frequency of the current transients decrease. This trend is most apparent for the longer 25-mer. We interpret these results as follows. When ssDNA is immobilized at high concentration on the PtNPs, the surface sites on the NP required for electrocatalytic N2H4 oxidation are blocked. This leads to lower and fewer ECA signals. In contrast, naked PtNPs are able to transfer electrons to UMEs having sparse coatings of ssDNA.

Mono- and dibasic phthalate bases (HP– and P2–) interact with 1,4-hydroquinone (1,4-H2Q) in acetonitrile through hydrogen bonding to form molecular complexes that oxidize at easier (less positive) potentials than pure 1,4-H2Q. The relatively large decrease in the oxidation overpotential for 1,4-H2Q, observed upon introducing these Brönsted bases, is an example of thermodynamic control of a PCET achieved with weak bases. The favoring effect occurs because the bases accept the protons released by 1,4-H2Q during its two-electron oxidation. Similar effects are observed when the phthalate bases were attached to the electrode surface. The hydrogen bonding was verified and measured by determining diffusion coefficients using 1H NMR spectroscopy. The modification of glassy carbon electrodes with phthalate diazonium salts was monitored by X-ray photoelectron spectroscopy. The experimental electrochemical responses by cyclic voltammetry were fitted to simulations following mechanisms previously reported and using the diffusion coefficients determined by 1H NMR.