Scanning Electrochemical Microscopy Tip Research Articles

Triple-function carbon-based Ca2+ ion-selective pH ring microelectrode to study real-time bacteria-mediated hydroxyapatite corrosion

BackgroundThe local pH change mediated by the pathogenic bacterial species Streptococcus mutans plays a significant role in the corrosion of hydroxyapatite (HA) present in the tooth in the dynamic oral cavity. The acid produced by the bacteria decreases the local pH and releases Ca2+ ions from the HA. We studied the bacteria-mediated demineralization of HA by scanning electrochemical microscopy (SECM) after growing S. mutans biofilm on HA for 7 days. ResultsWe notably developed a triple-function SECM-compatible tip that could be positioned above the biofilm. It can also measure the pH and [Ca2+] change simultaneously above the biofilm-HA substrate. The triple-function SECM tip is a combination of a potentiometric pH sensor deposited with iridium oxide and a dual-function carbon-based Ca2+ ion-selective membrane electrode with a slope of 67 mV/pH and 34.3 mV/log [Ca2+], respectively. The distance-controlled triple-function SECM tip monitored real-time pH and [Ca2+] changes 30 μm above the S. mutans biofilm. The high temporal resolution pH data demonstrated that after approximately 20 min of sucrose addition, S. mutans started to produce acid to titrate the solution buffer, causing a pH change from 7.2 to 6.5 for HA and from 7.2 to 5 for the glass substrate. We observed that, after 30 min of acid production, ∼300 μM of Ca2+ ions were increased at pH 6.5 above the biofilm surface as a result of the pH change in the local microenvironment. After the release of Ca2+ from HA, the pH environment again shifted toward the neutral side, from 6.5 to 7.2. Therefore, precipitation of Ca2+ happens at the top of the biofilm, thus corroding the HA from underneath. For a glass substrate, in contrast, no Ca2+ ions were released, and the pH did not change back to 7.2. We were able to observe the dynamics of the HA demineralization–remineralization process simultaneously with our newly developed triple-function SECM tip or microprobe. SignificanceThis technique could notably advance the study of similar complex processes, such as bacteria-mediated corrosion in biomedical and environmental contexts.

Investigation of the Electrocatalytic Reduction of Peroxydisulfate Using Scanning Electrochemical Microscopy.

The elementary steps of the electrocatalytic reduction of S2O82- using the Ru(NH3)63+/2+ redox couple were investigated using scanning electrochemical microscopy (SECM) and steady-state voltammetry (SSV). SECM investigations were carried out in a 0.1 M KCl solution using a 3.5 μm radius carbon ultramicroelectrode (UME) as the SECM tip and a 25 μm radius platinum UME as the substrate electrode. Approach curves were recorded in the positive feedback mode of SECM by reducing Ru(NH3)63+ at the tip electrode and oxidizing Ru(NH3)62+ at the substrate electrode, as a function of the tip-substrate separation and S2O82- concentration. The one-electron reaction between electrogenerated Ru(NH3)62+ and S2O82- yields the unstable S2O83•-, which rapidly dissociates to produce highly oxidizing SO4•-. Because SO4•- is such a strongly oxidizing species, it can be further reduced at both the tip and the substrate, or it can react with Ru(NH3)62+ to regenerate Ru(NH3)63+. SECM approach curves display a complex dependence on the tip-substrate distance, d, due to redox mediation reactions at both the tip and the substrate. Finite element method (FEM) simulations of both SECM approach curves and SSV confirm a previously proposed mechanism for the mediated reduction of S2O82- using the Ru(NH3)63+/2+ redox couple. Our results provide a lower limit for dissociation rate constant of S2O83•- (∼1 × 106 s-1), as well as the rate constants for electron transfer between SO4•- and Ru(NH3)62+ (∼1 × 109 M-1 s-1) and between S2O82- and Ru(NH3)62+ (∼7 × 105 M-1 s-1).

Voltammetric Detection of Singlet Oxygen Enabled by Nanogap Scanning Electrochemical Microscopy.

Despite the significance of singlet oxygen (1O2) in several biological, chemical, and energy storage systems, its voltammetric reduction at an electrode remains unreported. We address this issue using nanogap scanning electrochemical microscopy (SECM) in substrate-generation/tip-collection mode. Our investigation reveals a reductive process on the SECM tip at -1.0 V (vs Fc+/Fc) during the breakdown of the Li2CO3 substrate in deuterated acetonitrile. Notably, this value is approximately 0.9 V more positive than the reduction potential of triplet oxygen (3O2), consistent with thermodynamic estimates for the energy of the formation of 1O2. This finding holds significant implications for understanding the reaction mechanisms involving 1O2 in nonaqueous media.

Transient theory for scanning electrochemical microscopy of biological membrane transport: uncovering membrane-permeant interactions.

Scanning electrochemical microscopy (SECM) has emerged as a powerful method to quantitatively investigate the transport of molecules and ions across various biological membranes as represented by living cells. Advantageously, SECM allows for the in situ and non-destructive imaging and measurement of high membrane permeability under simple steady-state conditions, thereby facilitating quantitative data analysis. The SECM method, however, has not provided any information about the interactions of a transported species, i.e., a permeant, with a membrane through its components, e.g., lipids, channels, and carriers. Herein, we propose theoretically that SECM enables the quantitative investigation of membrane-permeant interactions by employing transient conditions. Specifically, we model the membrane-permeant interactions based on a Langmuir-type isotherm to define the strength and kinetics of the interactions as well as the concentration of interaction sites. Finite element simulation predicts that each of the three parameters uniquely affects the chronoamperometric current response of an SECM tip to a permeant. Significantly, this prediction implies that all three parameters are determinable from an experimental chronoamperometric response of the SECM tip. Complimentarily, the steady-state current response of the SECM tip yields the overall membrane permeability based on the combination of the three parameters. Interestingly, our simulation also reveals the optimum strength of membrane-permeant interactions to maximize the transient flux of the permeant from the membrane to the tip.

Evaluation of the Applicability of Voltammetric Modes in Scanning Electrochemical Microscopy for In Situ Corrosion Characterisation of Copper-Based Materials

Chemical imaging of corrosion processes involving copper species using scanning electrochemical microscopy has been hampered by the lack of soluble oxidation states for copper that can be achieved by amperometric conversion at the tip. Indeed, the only possibility is to reduce the corrosion products at the tip, thus modifying the chemical response of the electrode material and requiring subsequent redissolution of the copper deposits. Consequently, the limitations arising from the system prevented a full-scale quantification, requiring the development of new methodologies or the optimisation of those currently available, as we pursued with the present work. Therefore, the voltammetric behaviours of gold macro- and microelectrodes were evaluated with respect to the collection and redissolution of Cu2+ ions, with the aim of using them as sensing probes in scanning electrochemical microscopy (SECM) to investigate the activity of copper surfaces in acidic chloride-containing environments. Cyclic and square-wave voltammetric techniques were explored for copper collection and subsequent stripping on Au microelectrode tips in SECM with the objective to capture in situ image electrochemical reactivity distributions across copper surfaces undergoing corrosion.

Novel dynamic method based on CV-SECM and SWV-SECM for the in situ chemical imaging of reactive metal surfaces undergoing corrosion and corrosion inhibition illustrated with the case of copper corrosion inhibition by benzotriazole

A novel dynamic method for the chemical imaging of actively corroding metal surfaces using scanning electrochemical microscopy (SECM) is proposed by using CV-SECM and SWV-SECM. This consists of an adequate coupling of a repetitive voltammetric operation at the SECM tip while conducting the rastering routine for scanning a reactive copper surface, as well as its inhibition by benzotriazole. In this method, gold microelectrodes are employed as SECM tips, and therefore their electrochemical behaviour towards collection and redissolution of Cu2+ ions from synthetic solutions containing Cu2+ ions. Next, the dynamic application of voltammetric methods for the collection and stripping of copper on the Au probes while mapping copper surfaces was investigated. The proof of concept first involved continuous cyclic voltammetry (CV-SECM) in an acidified NaCl solution, in terms of Cu2+ dissolution and surface redox activity (with SECM in the feedback mode). Then, square wave voltammetry (SWV-SECM) was used to detect small copper dissolution from corroding and inhibitor-protected metal surfaces. The results are consistent with a heterogeneous nature of copper degradation and the development of protective films and passive layers.

Simultaneous Scanning Electrochemical Microscopy and UV-Vis Absorption Spectroelectrochemistry.

The combination of instrumental techniques allows obtaining precise and reliable information about the reactions taking place at the electrode/solution interface. Although UV-Vis absorption spectroelectrochemistry (UV-Vis SEC) provides a molecular insight about the species involved in the electrode process, obtaining information about the redox state of the products generated in this process is not always accessible by this technique. In this sense, scanning electrochemical microscopy (SECM) has a clear advantage, since it provides additional information on the oxidation state of the intermediates/products. Therefore, the combination of these two techniques facilitates obtaining a more complete picture of the electrochemical reaction studied from two different points of view, but under exactly the same experimental conditions. In this work, the combination of UV-Vis SEC in parallel configuration and SECM is carried out for the first time. This new technique allows distinguishing between those species that are electrochemically active and, at the same time, exhibit changes in the UV-Vis absorption spectra during the electrochemical reaction. The new experimental setup is first validated using ferrocenemethanol as a standard probe, concomitantly obtaining spectroscopic and electrochemical information that accurately describes the oxidation process. Finally, the strength of this combined technique is demonstrated by studying the antioxidant activity of o-vanillin (o-HVa) in the presence of electrogenerated superoxide. The information extracted from the new UV-Vis SEC/SECM technique makes it possible to identify, beyond any doubt, not only the origin of the electrochemical signals recorded in the SECM tip but also to evaluate the antioxidant effect of o-HVa at different concentrations.

Simultaneous Chemical Mapping of Live Biofilm Microenvironmental pH and Hydrogen Peroxide in Real Time with a Triple Scanning Electrochemical Microscopy Tip.

Dental plaque biofilm is a complex ecosystem. The distribution of microbial species in the biofilm is heavily influenced by local chemical interactions that result from diverse metabolic activities and the nature of the released molecules. As a relevant example, H2O2-producing bacteria can antagonize disease-associated bacteria, leading to the maintenance of a healthy oral microbiome. Herein, we report the development of a triple-sensor (redox, pH, and H2O2) scanning electrochemical microscopy (SECM) tip capable of simultaneously mapping the pH and H2O2 concentration produced by a dental plaque-derived multispecies biofilm grown on hydroxyapatite. The pH sensor of the triple SECM tip showed a near Nernstian slope of -71.1 ± 2 mV/pH (N = 3), whereas the H2O2 sensor showed a slope of -0.052 ± 0.002 nA/μM H2O2 at pH 7.2 and a detection limit of 1.0 ± 0.2 μM (N = 7). There is no significant difference in the sensitivities of H2O2 sensors at pH 6.2, 7.2, and 8.2 at 95% CI (N = 7). The pH and H2O2 sensors demonstrated excellent reversibility with response times of 3 and 5 s, respectively, along with reliable stability over 4 h at 37 °C. The sensors did not show any cross talk between pH and H2O2 concentration ([H2O2]) measurements, highlighting the accuracy and versatility of the SECM tip. Simultaneous chemical imaging of pH and [H2O2] across the biofilm revealed a clustered distribution of local H2O2 concentrations, ranging from 0 to 17 μM. Conversely, the local pH remained constant at 7.2. The relation of local chemical profiles and the distribution of bacterial species within the oral microbiome was experimentally investigated in the context of bacterial H2O2 antagonism. The benefit of clustered H2O2 production was that the total area of H2O2 produced by smaller clusters was 67% more than that of a single cluster with the same starting number of bacteria. Thus, this triple SECM tip can potentially be used to study local molecular mechanisms that result in dysbiosis of the oral microbiome.

Electrocatalysis of the hydrogen oxidation reaction on a platinum-decorated nanoporous gold surface studied by scanning electrochemical microscopy

The electrocatalytic activity of a platinum decorated nanoporous gold (Pt-NPG) surface was examined toward the Hydrogen Oxidation Reaction (HOR) using the feedback mode of Scanning Electrochemical Microscopy (SECM). In this system, the proton (H+) reduction and H2 oxidation reactions take place at the tip and the conductive substrate, respectively. Since the H2 oxidation process is highly surface sensitive, the positive feedback current is only observed when the substrate has an excellent electrocatalytic activity to oxidize H2 back to H+. A Pt microelectrode was used as an SECM tip and bare Pt, bare Au, NPG (Nanoporous gold), and Pt-NPG as substrate materials. Positive feedback currents from approach curves were observed for bare Pt and Pt-NPG surfaces, whereas hindered diffusion was noticed for Au and NPG surfaces. The obtained results confirm that the NPG surface is inactive for H2 oxidation, but it becomes active after decoration with a small amount of Pt.

An isolated single-particle-based SECM tip interface for single-cell NO sensing

As a key factor in cellular metabolic processes, nitric oxide (NO) is a challenging target for in situ real-time monitoring due to its transient property and short diffusion distance. Scanning electrochemical microscopy (SECM) has unique advantages in single-cell analysis, which can obtain the electrochemical current by scanning the cell surface with a tip microelectrode. In particular, it can further improved the electrochemical response by enhancing the interface properties of its tip. Here, an interface design strategy based on platinum single nanoparticle (Pt NP) was developed, and fluorinated self-assembled monolayers (SAMs) were used to further improve its performance. This modified tip was used as an SECM probe for NO concentration monitoring and morphological imaging of single MCF-7 cells. It has the high sensitivity (164.7 μA/μM·cm2) and good selectivity for NO detection, which benefits from the efficient catalytic properties of Pt NPs and high mass transport and hydrophobic antifouling properties of the interface. Notably, it shows a superior performance in detecting the fluctuation of NO released by a single MCF-7-cell under the stimulation of cadmium (Cd), which demonstrates a promising method for using a single-particle-based tip in SECM applications.

Ionic strength induced local electrodeposition of ZnO nanoparticles

The controlled maskless deposition of nanomaterials locally on surfaces is challenging and of relevance to numerous fields and applications, such as sensing and catalysis. Forming patterns of pre-synthesized, and therefore, well-defined NPs should become a general and simple building approach for manipulating surfaces. Electrochemical deposition of NPs under mild potentials is not common and is based on destabilizing nanomaterial dispersions by an electrical field. This approach that is termed from “nano to nano” is used here for the local deposition of ZnO NPs. Specifically, we employ scanning electrochemical microscopy (SECM) as a means of increasing locally the ionic strength by generating a flux of AuCl4− in a ZnO nanoparticle dispersion. The gold ions bind instantaneously to the ZnO NPs and drive their local deposition on a biased indium tin oxide (ITO) surface to form ZnO nanoparticle patterns embedded in Au. The ratio between Au and ZnO can be changed by controlling the AuCl4− flux at the SECM tip and the ITO potential. The as-electrodeposited ZnO NPs are characterized by microscopy and spectroscopy. Furthermore, the micron-sized Au-ZnO patterns are used as recyclable surface-enhanced Raman scattering (SERS) active substrates for the sensitive detection of a dye. The latter is only an example of the potential of such local nanomaterial-based patterns that can be readily formed by our approach.

Probing the Effects of Defects in Ultrathin Oxide Overlayers on the Selectivity of Encapsulated Electrocatalysts

Application of ultrathin oxide encapsulation layers on co-catalysts employed in z-scheme photocatalysis has the potential to increase overall efficiency through the prevention of undesirable back reactions and increased charge separation. [1,2] However, defects within the semipermeable oxide coatings – such as pinholes, cracks or particle protrusions – have been postulated to facilitate locally high rates of undesirable reactions by creating pathways for facile transport of undesired reactants to exposed active sites.[3] Local probe measurements, such as scanning electrochemical microscopy (SECM), can estimate the relative rates of production of product species generated from competing electrochemical reactions with high spatial resolution over the electrode. This can be leveraged to determine the influence of local defects on global performance metrics, such as the apparent permeability of the overlayer and selectivity of the electrode. In this presentation, we report the use of SECM to determine the influence of overlayer defects on the performance of a model silicon oxide (SiOx) -encapsulated Pt thin film electrocatalyst when operated under conditions where two competing reactions can occur. Motivated by Z-scheme photocatalysis, the hydrogen evolution reaction (HER) and Fe(III)/Fe(II) redox reaction were studied. After introducing new methodology to determine local selectivity towards HER using SECM tip current, the resulting selectivity maps are compared against defects seen in optical images, scanning electron micrographs, and atomic force microscopy images. This analysis reveals that that certain types of defects in the oxide overlayer can be responsible for a large percentage of the partial current density towards the undesired Fe(III) reduction reaction. Correcting for the defect contributions to the undesired reaction, it is determined that the true Fe(III) permeability values for the SiOx overlayers are almost two orders of magnitude lower than permeabilities determined from conventional analysis that ignored the presence of defects. Finally, different types of defects were studied revealing that the defect morphology can have varying influence on both the selectivity and calculated permeability. This work highlights the need for local measurements in addition to macroscopic measurements of oxide encapsulated catalysts as they are applied to more complex geometries such as particle photocatalysts.

Combined SECM and spectroscopy investigation of the interfacial chemistry of chalcopyrite during anodic oxidation

Hydrometallurgical copper extraction from chalcopyrite (CuFeS2), the most abundant copper-containing resource in the world, is always challenged by its passivation during industrial application, whose mechanisms are still not fully understood. In this study, the interfacial chemistry of chalcopyrite during the anodic oxidation was studied by combined Scanning Electrochemical Microscopy (SECM) and spectroscopic analyzes of Raman spectroscopy and X-ray absorption near-edge spectroscopy (XANES). Ex-situ SECM feedback mode was firstly used to detect and image the formation of passivating films. Then, the in-situ SECM substrate generation-tip collection (SG-TC) experiment by positioning the micro-platinum probe just above the chalcopyrite surface was conducted so that electrochemically active dissolution products can be deposited and monitored. Subsequently, Raman spectroscopy was performed on the SECM tip and the chalcopyrite substrate electrode respectively to identify the products that were released into the solution and then collected on the tip, as well as the ones left behind on the mineral surface. The SECM feedback mode mapping and approach curves demonstrated that under the passive and trans-passive potential regions, low conductive passivation films formed on the chalcopyrite surface, which contributed to the hindered dissolution of chalcopyrite due to the decreased electron transfer rate. In-situ SECM SG-TC results from the anodically oxidized chalcopyrite suggested that soluble sulfur species, most likely thiosulfate ions (S2O32−) were dissolved from the surface during the passivation films formation. The released S2O32− was evidenced by comparing with the pyrite anodic oxidation experiment and sulfur K-edge XANES analysis of the electrolytes collected after chalcopyrite anodic dissolution. In-situ Raman spectra of the oxidized chalcopyrite surface confirmed metal deficient layers were the passivation layers.

In Situ Monitoring of Extracellular K+ Using the Potentiometric Mode of Scanning Electrochemical Microscopy with a Carbon-Based Potassium Ion-Selective Tip.

The expression of potassium channels can be related to the occurrence and development of tumors. Their change would affect K+ outflow. Thus, in situ monitoring of extracellular K+ shows a great significance. Herein, the dual-functional K+ ion-selective electrode as the scanning electrochemical microscopy (SECM) tip (K+-ISE SECM tip) has been developed for in situ monitoring of the extracellular K+. Based on multi-wall carbon nanotubes as a transduction layer, the K+-ISE SECM tip realizes both the plotting of approach curves to position the tip for in situ detection and the recording of potential responses. It shows a near Nernstian response, good selectivity, and excellent stability. Based on these characteristics, it was used to in situ monitor K+ concentrations ([K+]o) of three breast cancer cell lines (MCF-7, MDA-MB-231, and SK-BR-3 cells) at 3 μm above the cell, and [K+]o of MDA-MB-231 cells show the highest value, followed by MCF-7 cells and SK-BR-3 cells. K+ outflow induced by electrical stimulation or pH changes of the culture environment (Δ[K+]o) was further determined, and the possible mechanism of K+ outflow was investigated with 4-aminopyridin (4-AP). MCF-7 cells present the largest value of Δ[K+]o, followed by MDA-MB-231 cells and SK-BR-3 cells at all the stimulation potentials, and pH 6.50 shows the greatest impact on K+ outflow of the three cell lines. The pretreatment of 4-AP changed K+ outflow, probably due to the regulation of voltage-gated channels. These findings provide insight into a deep understanding of the microenvironment influence on K+ outflow, thereby reflecting the possible mechanism of potassium channels.

Transparent Ultramicroelectrodes for Studying Interfacial Charge-Transfer Kinetics of Photoelectrochemical Water Oxidation at TiO2 Nanorods with Scanning Electrochemical Microscopy

Scanning electrochemical microscopy (SECM) has been extensively applied to the electrochemical analysis of the surfaces and interfaces of a photoelectrochemical (PEC) system. A semiconductor photoelectrode with a well-defined geometry and active surface area comparable to SECM's tip is highly desired for accurately quantifying interfacial charge-transfer activities and photoelectrochemically generated redox species, where the broadening effects due to the mass transfer gradient and nonlocal electron transfer at a planar semiconductor surface can be minimized. Here, we present a newly developed platform as a SECM substrate for investigating semiconductor PEC activities, which is based on a transparent ultramicroelectrode (UME) fabricated by using two-step photolithographic patterning and ion milling methods. This transparent UME with a 25 μm recessed disk shape is fully characterized with SECM for quantifying the interfacial charge-transfer rates of IrCl62-/IrCl63- by comparing with theoretical results from finite element simulations in COMSOL Multiphysics. When coated with TiO2 nanorods as a model semiconductor material, the transparent UME can be used to quantify the catalytic PEC water oxidation in a feedback mode of SECM by sampling tip and substrate current signals simultaneously. This transparent UME-SECM study provides insights into the potential-dependent PEC water oxidation reaction mechanism and the quantitative analysis of photocurrent contributions from water oxidation and the SECM tip-generated redox mediator. The transparent UME-SECM method can be potentially expanded to other SECM operation modes such as surface interrogation for understanding the dynamics of the electrode surfaces and interfaces of a PEC system.

Probing Activities of Individual Catalytic Nanoflakes by Tunneling Mode of Scanning Electrochemical Microscopy

The tunneling mode of scanning electrochemical microscopy (SECM) was developed recently and applied to studies of charge-transfer reactions at single metal nanoparticles (NPs). When an SECM tip is ...

PH Micro-Sensor from Irox SECM Microelectrode for Local pH Measurement While Chromium Electrodeposition

In this work, it will be presented the chromium electrodeposition mechanisms with the use of a Scanning ElectroChemical Microscopy (SECM) micro-electrode. Among several factors, the pH is key to achieve even a metallic deposit. pH buffers and complexing agents are often used for this purpose [1] but in the vicinity of the electrode, at the electrode/electrolyte interface, strong variations are highly likely to occur. To this respect, since pH is a key parameter, a pH micro-electrode is intended to be developed to scan the pH gradient upon the electrodeposition of chromium. The first part of this work is so devoted to the pH micro-electrode realization from microelectrode on which iridium oxide is deposited. Iridium oxide has been reported to be suitable for local pH sensing [2-4]. Different parameters have been studied such as IrOx thickness and thermal treatment. The addition of Nafion has been assessed as well to achieve a tradeoff between fast response time, good selectivity and good stability [5]. An optimized IrOx micro-electrode has been obtained and used as SECM micro-electrode to get the evolution of the pH at the electrode/electrolyte interface upon electrodeposition. Different micro-electrode diameters have been used to improve the resolution. These results are promising and will allow going further to the development of chromium electrodeposition electrolyte helping to select the different complexing agents.[1] D. Del Pianta et al. Determination of the chromium(III) reduction mechanism during chromium electroplating. Electrochimica Acta (2018) 234-241.[2] H. A. Elsen et al. Effects of electrodeposition conditions and protocol on the properties of iridium oxide pH sensors electrodes. J. Electrochem Soc. 156 (2009).[3] C. S. Santos et al. Fabrication and use od dual-function iridium oxide coated gold SECM tips. An application to pH monitoring above copper electrode surface during nitrate reduction. Electroanalysis 28 (2016) 1441-1447.[4] Z. Zhu et al. A fabrication of iridium oxide film pH micro-sensor on Pt ultramicroelectrode and its application on in-situ pH distribution of 316 stainless steel corrosion at open circuit potential. Sensors and Actuators B 25 (2018) 1974-1982.[5] I. G. Borislav et al. Thin-film IrOx pH microelectrode for microfluidic-based microsystems. Biosensors and Bioelectronics 21 (2005) 248-256.

Time-Resolved Local pH Measurements during CO2 Reduction Using Scanning Electrochemical Microscopy: Buffering and Tip Effects

The electrochemical reduction of CO2 is widely studied as a sustainable alternative for the production of fuels and chemicals. The electrolyte’s bulk pH and composition play an important role in the reaction activity and selectivity and can affect the extent of the buildup of pH gradients between the electrode surface and the bulk of the electrolyte. Quantifying the local pH and how it is affected by the solution species is desirable to gain a better understanding of the CO2 reduction reaction. Local pH measurements can be realized using Scanning Electrochemical Microscopy (SECM); however, finding a pH probe that is stable and selective under CO2 reduction reaction conditions is challenging. Here, we have used our recently developed voltammetric pH sensor to perform pH measurements in the diffusion layer during CO2 reduction using SECM, with high time resolution. Using a 4-hydroxylaminothiophenol (4-HATP)/4-nitrosothiophenol (4-NSTP) functionalized gold ultramicroelectrode, we compare the local pH developed above a gold substrate in an argon atmosphere, when only hydrogen evolution is taking place, to the pH developed in a CO2 atmosphere. The pH is monitored at a fixed distance from the surface, and the sample potential is varied in time. In argon, we observe a gradual increase of pH, while a plateau region is present in CO2 atmosphere due to the formation of HCO3– buffering the reaction interface. By analyzing the diffusion layer dynamics once the sample reaction is turned “off”, we gain insightful information on the time scale of the homogeneous reactions happening in solution and on the time required for the diffusion layer to fully recover to the initial bulk concentration of species. In order to account for the effect of the presence of the SECM tip on the measured pH, we performed finite element method simulations of the fluid and reaction dynamics. The results show the significant localized diffusion hindrance caused by the tip, so that in its absence, the pH values are more acidic than when the tip is present. Nonetheless, through the simulation, we can account for this effect and estimate the real local pH values across the diffusion layer.

High-Content Label-Free Single-Cell Analysis with a Microfluidic Device Using Programmable Scanning Electrochemical Microscopy.

The cellular heterogeneity and plasticity are often overlooked due to the averaged bulk assay in conventional methods. Optical imaging-based single-cell analysis usually requires specific labeling of target molecules inside or on the surface of the cell membrane, interfering with the physiological homeostasis of the cell. Scanning electrochemical microscopy (SECM), as an alternative approach, enables label-free imaging of single cells, which still confronts the challenge that the long-time scanning process is not feasible for large-scale analysis at the single-cell level. Herein, we developed a methodology combining a programmable SECM (P-SECM) with an addressable microwell array, which dramatically shortened the time consumption for the topography detection of the micropits array occupied by the polystyrene beads as well as the evaluation of alkaline phosphatase (ALP) activity of the 82 single cells compared with the traditional SECM imaging. This new arithmetic was based on the line scanning approach, enabling analysis of over 900 microwells within 1.2 h, which is 10 times faster than conventional SECM imaging. By implementing this configuration with the dual-mediator-based voltage-switching (VSM) mode, we investigated the activity of ALP, a promising marker for cancer stem cells, in hundreds of tumor and stromal cells on a single microwell device. The results discovered that not only a higher ALP activity is presented in cancer cells but also the heterogeneous distribution of kinetic constant (kf value) of ALP activity can be obtained at the single-cell level. By directly relating large numbers of addressed cells on the scalable microfluidic device to the deterministic routing of the above SECM tip, our platform holds potential as a high-content screening tool for label-free single-cell analysis.

PH Micro-Sensor from IrOx SECM Microelectrode for Local pH Measurement While Chromium Electrodeposition

In this work, it will be presented the chromium electrodeposition mechanisms with the use of a Scanning ElectroChemical Microscopy (SECM) micro-electrode. Among several factors, the pH is key to achieve even a metallic deposit. pH buffers and complexing agents are often used for this purpose [1] but in the vicinity of the electrode, at the electrode/electrolyte interface, strong variations are highly likely to occur. To this respect, since pH is a key parameter, a pH micro-electrode is intended to be developed to scan the pH gradient upon the electrodeposition of chromium. The first part of this work is so devoted to the pH micro-electrode realization from microelectrode on which iridium oxide is deposited. Iridium oxide has been reported to be suitable for local pH sensing [2-4]. Different parameters have been studied such as IrOx thickness and thermal treatment. The addition of Nafion has been assessed as well to achieve a tradeoff between fast response time, good selectivity and good stability [5]. An optimized IrOx micro-electrode has been obtained and used as SECM micro-electrode to get the evolution of the pH at the electrode/electrolyte interface upon electrodeposition. Different micro-electrode diameters have been used to improve the resolution. These results are promising and will allow going further to the development of chromium electrodeposition electrolyte helping to select the different complexing agents.[1] D. Del Pianta et al. Determination of the chromium(III) reduction mechanism during chromium electroplating. Electrochimica Acta (2018) 234-241.[2] H. A. Elsen et al. Effects of electrodeposition conditions and protocol on the properties of iridium oxide pH sensors electrodes. J. Electrochem Soc. 156 (2009).[3] C. S. Santos et al. Fabrication and use od dual-function iridium oxide coated gold SECM tips. An application to pH monitoring above copper electrode surface during nitrate reduction. Electroanalysis 28 (2016) 1441-1447.[4] Z. Zhu et al. A fabrication of iridium oxide film pH micro-sensor on Pt ultramicroelectrode and its application on in-situ pH distribution of 316 stainless steel corrosion at open circuit potential. Sensors and Actuators B 25 (2018) 1974-1982.