Scanning Electrochemical Microscopy Tip Research Articles

Silver Particle Nucleation and Growth at Liquid/Liquid Interfaces: A Scanning Electrochemical Microscopy Approach

Scanning electrochemical microscopy (SECM) has been used to induce and monitor the electrodeposition of silver particles at a liquid/liquid interface by the electron transfer reaction between aqueous Ag+ ions, generated by anodic dissolution of an Ag disk ultramicroelectrode (UME), and bis(pentamethylcyclopentadienyl) iron (decamethylferrocene, DMFc) in a 1,2-dichloroethane (DCE) phase. A two-electrode system with an Ag UME as the SECM tip was used to investigate the factors affecting the deposition process, such as the tip-interface separation, potential applied to the tip, concentration of the reductant in the DCE phase, and the reaction driving force, which was controlled by the concentration ratio of a common ion (ClO4-) in the two phases. A theoretical model was developed and rate constants for Ag particle nucleation and growth at the water/DCE interface were obtained by thorough analysis of experimental current-time curves. It was found that Ag+ ion adsorption at the interface, coupled to particle nucleation and growth, best described the experimental data. The growth of Ag particles at the liquid/liquid interface was confirmed by independent microscopy measurements.

Determination of the Diffusion Coefficient of Monosaccharides with Scanning Electrochemical Microscopy (SECM)

AbstractThe principle of a simple time of flight (TOF) diffusion coefficient measuring method is presented. It is based on the special capability of Scanning Electrochemical Microscopy (SECM). Its applicability has been investigated experimentally in comparison with conventional electrochemical methods. Complex irreversible electrode processes usually bring in uncertainty into electrochemical diffusion coefficient measurements. One of these, the electrochemical oxidation of saccharides in basic media at a copper electrode, was selected for the studies. Microsize copper electrodes used also as SECM tip, and conventional size copper electrodes were used. Diffusion coefficients of D‐glucose, D‐arabinose, D‐ribose, and D‐galactose were measured. The results prove the advantages of the SECM‐TOF method in the case investigated.

Detection of hydrogen peroxide produced at a liquid/liquid interface using scanning electrochemical microscopy

Scanning electrochemical microscopy (SECM) was used to monitor in situ hydrogen peroxide (H 2O 2) produced at a polarized water/1,2-dichloroethane (DCE) interface. The water/DCE interface was formed between a DCE droplet containing decamethylferrocene (DMFc) supported on a solid electrode and an acidic aqueous solution. H 2O 2 was generated by reducing oxygen with DMFc at the water/DCE interface, and was detected with a SECM tip positioned in the vicinity of the interface using a substrate generation/tip collection mode. This work shows unambiguously how the H 2O 2 generation depends on the polarization of the liquid/liquid interface, and how proton-coupled electron transfer reactions can be controlled at liquid/liquid interfaces.

Scanning Electrochemical Microscopy of DNA Monolayers Modified with Nile Blue

Scanning electrochemical microscopy (SECM) is used to probe long-range charge transport (CT) through DNA monolayers containing the redox-active Nile Blue (NB) intercalator covalently affixed at a specific location in the DNA film. At substrate potentials negative of the formal potential of covalently attached NB, the electrocatalytic reduction of Fe(CN)6(3-) generated at the SECM tip is observed only when NB is located at the DNA/solution interface; for DNA films containing NB in close proximity to the DNA/electrode interface, the electrocatalytic effect is absent. This behavior is consistent with both rapid DNA-mediated CT between the NB intercalator and the gold electrode as well as a rate-limiting electron transfer between NB and the solution phase Fe(CN)6(3-). The DNA-mediated nature of the catalytic cycle is confirmed through sequence-specific and localized detection of attomoles of TATA-binding protein, a transcription factor that severely distorts DNA upon binding. Importantly, the strategy outlined here is general and allows for the local investigation of the surface characteristics of DNA monolayers both in the absence and in the presence of DNA binding proteins. These experiments highlight the utility of DNA-modified electrodes as versatile platforms for SECM detection schemes that take advantage of CT mediated by the DNA base pair stack.

Interrogation of Surfaces for the Quantification of Adsorbed Species on Electrodes: Oxygen on Gold and Platinum in Neutral Media

We introduce a new in situ electrochemical technique based on the scanning electrochemical microscope (SECM) operating in a transient feedback mode for the detection and direct quantification of adsorbed species on the surface of electrodes. A SECM tip generates a titrant from a reversible redox mediator that reacts chemically with an electrogenerated or chemically adsorbed species at a substrate of about the same size as the tip, which is positioned at a short distance from it (ca.1 microm). The reaction between the titrant and the adsorbate provides a transient positive feedback loop until the adsorbate is consumed completely. The sensing mechanism is provided by the contrast between positive and negative feedback, which allows a direct quantification of the charge neutralized at the substrate. The proposed technique allows quantification of the adsorbed species generated at the substrate at a given potential under open circuit conditions, a feature not attainable with conventional electrochemical methods. Moreover, the feedback mode allows the tip to be both the titrant generator and detector, simplifying notably the experimental setup. The surface interrogation technique we introduce was tested for the quantification of electrogenerated oxides (adsorbed oxygen species) on gold and platinum electrodes at neutral pH in phosphate and TRIS buffers and with two different mediator systems. Good agreement is found with cyclic voltammetry at the substrate and with previous results in the literature, but we also find evidence for the formation of "incipient oxides" which are not revealed by conventional voltammetry. The mode of operation of the technique is supported by digital simulations, which show good agreement with the experimental results.

Local electron transfer rate measurements on modified and unmodified glassy carbon electrodes

A natural and artificial distribution of electron transfer activity on glassy carbon electrodes can be observed and quantified by the use of scanning electrochemical microscopy (SECM). A large (sevenfold) spread in rate constant is found for randomly sampled sites on polished, untreated glassy carbon surfaces. Direct-mode oxidation with the SECM tip was used to produce small regions of oxidized carbon on a polished surface. A large increase in electron transfer rate for the Fe(II/III) ion is observed on the locally oxidized carbon surface in comparison to the unoxidized region. Rate constant measurements made along a line profiles the transition from unoxidized to oxidized surfaces. SECM images of defect sites show reaction–rate variations. Rate constants measured at several locations of the defective surface allows discrimination between the kinetic and topographic components of the SECM image.

Reading Oxidation States of Encapsulated Ferrocenium in Calix[4]arene Heterodimers Immobilized on Gold Dots by Means of a Scanning Electrochemical Microscopy Probe

The oxidation states of encapsulated ferrocenium (Fc+ filled in calix[4]arene heterodimers) in the form of a self-assembled monolayer (SAM) immobilized on a gold surface were recognized by a 2.2 μm diameter ultramicroelectrode (tip) in combination with scanning electrochemical microscopy (SECM). By approaching the SECM tip biased at −0.10 V to the vicinity of the encapsulated Fc+ moieties, an electrochemical current at the tip was observed, indicating the reduction of the encapsulated Fc+ to ferrocene (Fc). On the other hand, no Faradaic current was observed at the tip if the immobilized SAM with encapsulated Fc was beneath the same tip. When the immobilized Fc+ is alternated between the two oxidation states by scanning the electrochemical potential applied to the SAM between −0.20 and 0.50 V, an oxidation current appeared and disappeared periodically at the tip biased at 0.40 V, corresponding to the oxidation state exchange of the Fc+ beneath it. In this way, the SECM tip can be used to monitor or change the oxidation states of the encapsulated Fc+ in the SAM on the gold surface. Two different open circuit potential−time curves were observed for the two oxidation states in the SAM on the gold surface, which confirms that both oxidation states of the encapsulated Fc+ are stable. This system might be developed to a molecular memory device with a small SECM tip used as the reading and writing probe.

Screening of Oxygen Evolution Electrocatalysts by Scanning Electrochemical Microscopy Using a Shielded Tip Approach

Oxygen evolution electrocatalysts in acidic media were studied by scanning electrochemical microscopy (SECM) in the substrate generation-tip collection (SG-TC) imaging mode with a 100 microm diam tip. Pure IrO2 and Sn(1-x)Ir(x)O2 combinatorial mixtures were prepared by a sol-gel route to form arrays of electrocatalyst spots. The experimental setup has been developed to optimize screening of electrocatalyst libraries under conditions where the entire array is capable of the oxygen evolution reaction (OER). The activity of individual spots was determined by reducing the interference from the reaction products of neighboring spots diffusing to the tip over the spot of interest. A gold layer deposited on the external wall of the SECM tip was used as a tip shield. In this study the shield was kept at a constant potential to reduce oxygen under mass transfer controlled conditions. The tip shield consumes oxygen coming from the neighbor spots in the array and enables the tip to correctly detect the activity of the spot below the tip. Simulations and experimental results are shown, demonstrating the effectiveness of the tip shield with the SG-TC setup in determining the properties of the composite materials and imaging arrays.

Real-time detection of Cu 2+ sequestration and release by immobilized apo-metallothioneins using SECM combined with SPR

Scanning electrochemical microscopy (SECM) combined with surface plasmon resonance (SPR), SECM–SPR, was applied for real-time detection of the incorporation of Cu 2+ by apo-metallothionein (apo-MT) immobilized on the SPR substrate and release of Cu 2+ from surface-confined metallothionein (MT). Cu 2+ anodically stripped from a Cu-coated SECM Au tip was sequestered by apo-MT upon its diffusion to the SPR substrate, and release of Cu 2+ by MT was accomplished by generating protons via oxidation of hydroquinone at the tip. The high sensitivity of the SPR instrument is capable of following the structural and compositional changes of MT molecules during the metal sequestration and release processes. Due to the enhanced mass transfer rate at the SECM tip, the complication of mass transfer limitation on kinetic measurements, commonly encountered in flow injection SPR, is circumvented. The time-resolved SPR response reveals stepwise changes among three stable MT structures and allows the number of copper ions coordinated in each structure to be determined. The numbers of copper ions incorporated by each MT molecule in the three structures were determined to be 5, 9, and 12. This work expands the SECM–SPR approach to assessments of the dynamics and affinity of binding of small ions to surface-confined proteins and to studies of proteins that do not undergo facile electron transfer reactions.

SECM visualization of the spatial variability of enzyme-polymer spots. 3. Enzymatic feedback mode

The responses of a PQQ-GDH entrapped in a polymer structure to mixtures of glucose and maltose were evaluated. Each compound was considered in the concentration range of 0-0.2 mM. Imaging was performed at constant height in the enzymatic feedback mode of scanning electrochemical microscopy (SECM). The enzyme-polymer spot was discretized into 15 x 15 mum(2) substructures which were treated as independent individual microsensors. The response surfaces of the individual microsensors were approximated with a linear regression model. The coefficients in the derived equations represent contributions from topography, glucose concentration, maltose concentration, and the competition of glucose and maltose for the same active site of PQQ-GDH to the measured signal. The ratio of glucose and maltose contributions to the current at the SECM tip was constant for all microsensors and it was predominantly determined by the ratio of the turnover rates of both analytes in the PQQ-GDH catalyzed reaction. Using the difference between these coefficients, it was possible to select the microsensors within the overall enzyme-polymer spot that provided the best data for quantifying glucose and maltose by the artificial neural network used. The quantification of glucose and maltose was successful, except when the contributions from the components of the mixture were n (g)=k n units of glucose and simultaneously n (m)= 1.86(1-k)n units of maltose, for each constant n > 0 and k E <0,1>.

Selective Insulation with Poly(tetrafluoroethylene) of Substrate Electrodes for Electrochemical Background Reduction in Scanning Electrochemical Microscopy

We describe a wet process for the fabrication of poly(tetrafluoroethylene) (PTFE)-covered electrodes in which arrays of holes ( approximately 200 microm) are formed. The PTFE coating provides electrical insulation of most of the electrode surface with selected regions exposed for electrochemical experiments. The arrays of microholes can be controllably patterned and filled with precursor solutions using a piezoelectric dispenser. A micrometer spot of electrocatalyst is produced after reduction of the precursor. The application is tested for scanning electrochemical microscopy (SECM) in the tip generation-substrate collection (TG-SC) studies of electrocatalysts. The method is shown to reduce the substrate background currents that are included in the electrochemical signal read from the local perturbation induced with the SECM tip to the substrate in the TG-SC mode of SECM. This background current reduction is consistent with the decrease in the exposed area of the electrode. The general methodology for the fabrication of the substrate electrodes and two proof-of-concept applications in the TG-SC SECM modality are described.

Scanning electrochemical microscopy study of ion transfer process across water/2-nitrophenyloctylether interface supported by hydrophobic carbon ceramic electrode

A carbon ceramic electrode (CCE) modified with the redox probe—decamethylferrocene solution in hydrophobic organic solvent—2-nitrophenyloctyl ether and immersed into an aqueous solution was studied by scanning electrochemical microscopy (SECM). After the electrochemical oxidation of decamethylferrocene, its cations were detected near the electrode surface in the aqueous phase. This indicates that some fraction of the redox-active cations electrochemically produced in the organic phase is transferred across the liquid/liquid interface. They are reduced at the SECM tip and form a solid deposit. The amount of deposited decamethylferrocene was estimated by the anodic reaction at the tip. It is affected by the substrate–tip distance, deposition time, and electrolyte concentration. The SECM images of unmodified and modified CCEs are consistent with their heterogeneous structure.

Scanning Electrochemical Imprinting Microscopy: A Tool for Surface Patterning

An approach for patterning surface with different structures in a single step is demonstrated. The approach is based on the construction of nondisk shape microelectrodes combined with the feedback mode of scanning electrochemical microscopy (SECM) for generating an anisotropic flux of electroactive species. The latter attains the shape of the microelectrode and imprints its shape upon reaction with the surface. We have demonstrated this approach in two model systems. The etching of using electrogenerated bromine resulted in the formation of etching structures imprinting the shape of the Pt microelectrode on the . The second system comprised the electrogeneration of silver ions from a silver microelctrode, which reacted with a copper surface causing the deposition of silver patterns matching the shape of the silver microelectrode. SECM tips with different shapes were constructed by different methods, including electron-beam lithography.

Investigation of the interactions between silver nanoparticles and Hela cells by scanning electrochemical microscopy

The interactions between Hela cells and silver nanoparticles (AgNPs) have been studied by scanning electrochemical microscopy (SECM) with both IrCl(6)(2-/3-) and Fe(CN)(6)(3-/4-) as the dual mediators. IrCl(6)(2-), which can be produced in situ and react with AgNPs, is used as the mediator between the AgNPs on the cells and the SECM tip. Another redox couple, Fe(CN)(6)(3-/4-), which has a similar hydrophilicity to IrCl(6)(2-/3-), but cannot react with AgNPs, is also employed for the contrast experiments. The cell array is cultured successfully onto a Petri dish by microcontact printing (muCP) technique, which can provide a basic platform for studying of single cells. The approach curve and line scan are the two methods of SECM employed here to study the Hela cells. The former can provide the information about the interaction between Hela cells and AgNPs whereas the later gives the cell imaging. The permeability of cell membranes and morphology are two main factors which have effects on the feedback mode signals when K(3)Fe(CN)(6) is used as the mediator. The permeability of the cell membranes can be ignored after interaction with high concentration of AgNP solution and the height of the Hela cells is slightly decreased in this process. The kinetic rate constants (k(0)) between IrCl(6)(2-) and Ag on the Hela cell can be evaluated using K(3)IrCl(6) as the mediator, and they are increased with the higher concentrations of the AgNP solutions. The k(0) is changed about 10 times from 0.43 +/- 0.04 x 10(-4) to 1.25 +/- 0.07 x 10(-4) and to 3.93 +/- 1.9 x 10(-4) cm s(-1) corresponding to 0, 1 and 5 mM of AgNO(3) solution. The experimental results demonstrate that the AgNPs can be adsorbed on the cell surface and detected by SECM. Thus, the amount of AgNPs adsorbed on cell membranes and the permeability or morphology changes can be investigated simultaneously using this approach. The dual mediator system and cell array fabricated by muCP technique can provide better reproducibility because they can simplify experiments, and provide a platform for further single cell detection.

Electrochemical Imaging of Localized Sandwich DNA Hybridization Using Scanning Electrochemical Microscopy

Imaging of localized hybridization of nucleic acids immobilized on gold-DNA chip was performed by means of the feedback mode of scanning electrochemical microscopy (SECM). Thiol-tethered oligodeoxynucleotide (HS-ODN) probes, spotted on a gold surface, were hybridized with unmodified target sequence via sandwich hybridization with a biotinylated signaling probe. Spots where sequence-specific hybridization had occurred were developed by adding a streptavidin-alkaline phosphatase conjugate and biocatalyzed precipitation of an insoluble and insulating product. As a consequence, the surface conductivity of the spotted region of the chip where hybridization had taken place changed. These changes in conductivity were sensitively detected by the SECM tip. The proposed method allows imaging of a DNA array in a straightforward way. Analysis of real samples was also performed coupling this method with polymerase chain reaction. The imaging of 60 nM PCR amplicon (255 bp) was demonstrated.

Fabrication of Active Horseradish Peroxidase Micropatterns with a High Resolution by Scanning Electrochemical Microscopy

AbstractWe used a new reactive species OH− to fabricate active horseradish peroxidase (HRP) micropatterns with a high resolution by scanning electrochemical microscopy (SECM) coupled with a carbon fiber disk electrode as the SECM tip. In this method, except for active HRP micropatterns predesigned other regions on a HRP‐immobilized substrate were deactivated by OH− generated at the tip held at −1.7 V in 1.0 mol/L KCl containing 2.0×10−3 mol/L benzoquinone (BQ) (pH 8.0). The feedback mode of SECM with a tip potential of −0.2 V was used to characterize the active HRP micropatterns in 1.0 mol/L KCl containing 2.0×10−3 mol/L BQ and 2.0×10−3 mol/L H2O2.

Reactivity at the film/solution interface of ex situ prepared bismuth film electrodes: A scanning electrochemical microscopy (SECM) and atomic force microscopy (AFM) investigation

Bismuth film electrodes (BiFEs) prepared ex situ with and without complexing bromide ions in the modification solution were investigated using scanning electrochemical microscopy (SECM) and atomic force microscopy (AFM). A feedback mode of the SECM was employed to examine the conductivity and reactivity of a series of thin bismuth films deposited onto disk glassy carbon substrate electrodes (GCEs) of 3 mm in diameter. A platinum micro-electrode ( ϕ = 25 μm) was used as the SECM tip, and current against tip/substrate distance was recorded in solutions containing either Ru(NH 3) 6 3+ or Fe(CN) 6 4− species as redox mediators. With both redox mediators positive feedback approach curves were recorded, which indicated that the bismuth film deposition protocol associated with the addition of bromide ions in the modification solution did not compromise the conductivity of the bismuth film in comparison with that prepared without bromide. However, at the former Bi film a slight kinetic hindering was observed in recycling Ru(NH 3) 6 3+, suggesting a different surface potential. On the other hand, the approach curves recorded by using Fe(CN) 6 4− showed that both types of the aforementioned bismuth films exhibited local reactivity with the oxidised form of the redox mediator, and that bismuth film obtained with bromide ions exhibited slightly lower reactivity. The use of SECM in the scanning operation mode allowed us to ascertain that the bismuth deposits were uniformly distributed across the whole surface of the glassy carbon substrate electrode. Comparative AFM measurements corroborated the above findings and additionally revealed a denser growth of smaller bismuth crystals over the surface of the substrate electrode in the presence of bromide ions, while the crystals were bigger but sparser in the absence of bromide ions in the modification solution.

Scanning Electrochemical Microscopy. 58. Application of a Micropipet-Supported ITIES Tip To Detect Ag+ and Study Its Effect on Fibroblast Cells

We report the use of a micropipet-supported ITIES (interface between two immiscible electrolyte solutions, also called a liquid/liquid (L/L) or water/oil (W/O) interface) as a scanning electrochemical microscopy (SECM) tip to detect silver ion and explore Ag+ toxicity in living cells. A 1,2-dichloroethane solution containing a commercially available calixarene-based Ag+ ionophore (IV) was injected into a micrometer-size glass pipet to construct an Ag+-selective SECM tip. The local Ag+ concentration, down to the micromolar level, in the vicinity of living fibroblast cells, was monitored by SECM approach curves and through imaging of the uptake and efflux of Ag+ by living fibroblast cells in real time. The results show that several stages of interaction between Ag+ and fibroblast cells exist. Since a number of biological processes of cells are involved with non-redox-active ions, the work presented here provides a new way to explore cell metabolism, drug delivery, and toxicity assessment by SECM.

A unified new analytical approximation for negative feedback currents with a microdisk SECM tip

Abstract Analytical approximations for steady-state negative feedback currents in scanning electrochemical microscopy (SECM) experiments using a microdisk are discussed. A precise study of the limit behavior for small tip–substrate distances and all microdisks leads to an analytical expression without any fit. From this analytical limit a new mathematical function is proposed which is the first expression that accurately fit any negative feedback approach curve with only three adjustable parameters (for a given R g value, characterizing the insulator thickness of the microdisk electrode, between 1.001 and 1000). Finally a unified new analytical approximation of all negative feedback approach curves with tip–substrate distances and R g as variables is demonstrated. Except for very large R g values (over 200), which is not an experimental interesting configuration, the error is below ±0.01 on normalized current.

Constant-distance mode AC-SECM for the visualisation of corrosion pits

The local visualisation of corrosion pits has so far failed to convincingly prove the advantage of scanning microscopy in overcoming Abbey’s Limit. Thus, for imaging pit initiation and instant pit growth the resolution of the methods used for detecting the related changes in local electrochemical activity has to be significantly improved. Progress into this direction has been achieved by combining shear force based constant-distance mode scanning electrochemical microscopy (SECM) with the alternating current mode of this technique (AC-SECM). A very small corrosion pit was generated by inducing active corrosion underneath an accurately positioned 5 μm diameter Pt disk SECM tip that served as counter electrode in the direct mode of SECM. Before and after pit initiation, constant-distance mode AC-SECM images were recorded and subsequently used to identify the area of broken passivity. The results demonstrate the feasibility of the suggested approach to image actively corroding sites with overall dimensions of only a few micrometers. Combination of constant-distance scanning with AC-SECM is an important prerequisite for further improving spatial resolution by employing smaller-diameter micro- or even nano-electrodes as SECM tips.