Stochastic Electrochemical Measurement of a Biofouling Layer on Gold.

Adsorption of a biofouling layer on the surface of biosensors decreases the electrochemical activity and hence shortens the service life of biosensors, particularly implantable and wearable biosensors. Real-time quantification of the loss of activity is important for in situ assessment of performance while presenting an opportunity to compensate for the loss of activity and recalibrate the sensor to extend the service life. Here, we introduce an electrochemical noise measurement technique as a tool for the quantification of the formation of a biofouling layer on the surface of gold. The technique uniquely affords thermodynamic and kinetic information without applying an external bias (potential and/or current), hence allowing the system to be appraised in its innate state. The technique relies on the analysis of non-faradaic current and potential fluctuations that are intrinsically generated by the interaction of charged species at the electrode surface, i.e., gold. An analytical model is extended to explain the significance of parameters drawn from statistical analysis of the noise signal. This concept is then examined in buffered media in the presence of albumin, a common protein in the blood and a known source of a fouling layer in biological systems. Results indicate that the statistical analysis of the noise signal can quantify the loss of electrochemical activity, which is also corroborated by impedance spectroscopy as a complementary technique.

SARS-CoV-2 spike proteins react with Au and Si, are electrically conductive and denature at 3 × 108 V m-1: a surface bonding and a single-protein circuit study.

Developing means to characterise SARS-CoV-2 and its new variants is critical for future outbreaks. SARS-CoV-2 spike proteins have peripheral disulfide bonds (S-S), which are common in all spike proteins of SARS-CoV-2 variants, in other types of coronaviruses (e.g., SARS-CoV and MERS-CoV) and are likely to be present in future coronaviruses. Here, we demonstrate that S-S bonds in the spike S1 protein of SARS-CoV-2 react with gold (Au) and silicon (Si) electrodes. Bonding to Si is induced by a spontaneous electrochemical reaction that involves oxidation of Si-H and the reduction of the S-S bonds. The reaction of the spike protein with Au enabled single-molecule protein circuits, by connecting the spike S1 protein between two Au nano-electrodes using the scanning tunnelling microscopy-break junction (STM-BJ) technique. The conductance of a single spike S1 protein was surprisingly high and ranged between two states of 3 × 10-4 G 0 and 4 × 10-6 G 0 (1G 0 = 77.5 µS). The two conductance states are governed by the S-S bonds reaction with Au which controls the orientation of the protein in the circuit, and via which different electron pathways are created. The 3 × 10-4 G 0 level is attributed to a single SARS-CoV-2 protein connecting to the two STM Au nano-electrodes from the receptor binding domain (RBD) subunit and the S1/S2 cleavage site. A lower 4 × 10-6 G 0 conductance is attributed to the spike protein connecting to the STM electrodes from the RBD subunit and the N-terminal domain (NTD). These conductance signals are only observed at electric fields equal to or lower than 7.5 × 107 V m-1. At an electric field of 1.5 × 108 V m-1, the original conductance magnitude decreases accompanied by a lower junction yield, suggesting a change in the structure of the spike protein in the electrified junction. Above an electric field of 3 × 108 V m-1, the conducting channels are blocked and this is attributed to the spike protein denaturing in the nano-gap. These findings open new venues for developing coronavirus-capturing materials and offer an electrical method for analysing, detecting and potentially electrically deactivating coronaviruses and their future variants.

Advances in single-molecule junctions as tools for chemical and biochemical analysis.

The development of miniaturized electronics has led to the design and construction of powerful experimental platforms capable of measuring electronic properties to the level of single molecules, along with new theoretical concepts to aid in the interpretation of the data. A new area of activity is now emerging concerned with repurposing the tools of molecular electronics for applications in chemical and biological analysis. Single-molecule junction techniques, such as the scanning tunnelling microscope break junction and related single-molecule circuit approaches have a remarkable capacity to transduce chemical information from individual molecules, sampled in real time, to electrical signals. In this Review, we discuss single-molecule junction approaches as emerging analytical tools for the chemical and biological sciences. We demonstrate how these analytical techniques are being extended to systems capable of probing chemical reaction mechanisms. We also examine how molecular junctions enable the detection of RNA, DNA, and traces of proteins in solution with limits of detection at the zeptomole level.

On-Surface Azide-Alkyne Cycloaddition Reaction: Does It Click with Ruthenium Catalysts?

Owing to its simplicity, selectivity, high yield, and the absence of byproducts, the "click" azide-alkyne reaction is widely used in many areas. The reaction is usually catalyzed by copper(I), which selectively produces the 1,4-disubstituted 1,2,3-triazole regioisomer. Ruthenium-based catalysts were later developed to selectively produce the opposite regioselectivity─the 1,5-disubstituted 1,2,3-triazole isomer. Ruthenium-based catalysis, however, remains only tested for click reactions in solution, and the suitability of ruthenium catalysts for surface-based click reactions remains unknown. Also unknown are the electrical properties of the 1,4- and 1,5-regioisomers, and to measure them, both isomers need to be assembled on the electrode surface. Here, we test whether ruthenium catalysts can be used to catalyze surface azide-alkyne reactions to produce 1,5-disubstituted 1,2,3-triazole, and compare their electrochemical properties, in terms of surface coverages and electron transfer kinetics, to those of the compound formed by copper catalysis, 1,4-disubstituted 1,2,3-triazole isomer. Results show that ruthenium(II) complexes catalyze the click reaction on surfaces yielding the 1,5-disubstituted isomer, but the rate of the reaction is remarkably slower than that of the copper-catalyzed reaction, and this is related to the size of the catalyst involved as an intermediate in the reaction. The electron transfer rate constant (ket) for the ruthenium-catalyzed reaction is 30% of that measured for the copper-catalyzed 1,4-isomer. The lower conductivity of the 1,5-isomer is confirmed by performing nonequilibrium Green's function computations on relevant model systems. These findings demonstrate the feasibility of ruthenium-based catalysis of surface click reactions and point toward an electrical method for detecting the isomers of click reactions.

Effect of Electric Fields on Silicon-Based Monolayers.

Electric fields can induce bond breaking and bond forming, catalyze chemical reactions on surfaces, and change the structure of self-assembled monolayers on electrode surfaces. Here, we study the effect of electric fields supplied either by an electrochemical potential or by conducting atomic force microscopy (C-AFM) on Si-based monolayers. We report that typical monolayers on silicon undergo partial desorption followed by the oxidation of the underneath silicon at +1.5 V vs Ag/AgCl. The monolayer loses 28% of its surface coverage and 55% of its electron transfer rate constant (ket) when +1.5 V electrochemical potential is applied on the Si surface for 10 min. Similarly, a bias voltage of +5 V applied by C-AFM induces complete desorption of the monolayer at specific sites accompanied by an average oxide growth of 2.6 nm when the duration of the bias applied is 8 min. Current-voltage plots progressively change from rectifying, typical of metal-semiconductor junctions, to insulating as the oxide grows. These results define the stability of Si-based organic monolayers toward electric fields and have implication in the design of silicon-based monolayers, molecular electronics devices, and on the interpretation of charge-transfer kinetics across them.

Impermeable Graphene Oxide Protects Silicon from Oxidation.

The presence of a natural silicon oxide (SiOx) layer over the surface of silicon (Si) has been a roadblock for hybrid semiconductor and organic electronics technology. The presence of an insulating oxide layer is a limiting operational factor, which blocks charge transfer and therefore electrical signals for a range of applications. Etching the SiOx layer by fluoride solutions leaves a reactive Si-H surface that is only stable for few hours before it starts reoxidizing under ambient conditions. Controlled passivation of silicon is also of key importance for improving Si photovoltaic efficiency. Here, we show that a thin layer of graphene oxide (GOx) prevents Si surfaces from oxidation under ambient conditions for more than 30 days. In addition, we show that the protective GOx layer can be modified with molecules enabling a functional surface that allows for further chemical conjugation or connections with upper electrodes, while preserving the underneath Si in a nonoxidized form. The GOx layer can be switched electrochemically to reduced graphene oxide, allowing the development of a dynamic material for molecular electronics technologies. These findings demonstrate that 2D materials are alternatives to organic self-assembled monolayers that are typically used to protect and tune the properties of Si and open a realm of possibilities that combine Si and 2D materials technologies.

Ultrasonic Generation of Thiyl Radicals: A General Method of Rapidly Connecting Molecules to a Range of Electrodes for Electrochemical and Molecular Electronics Applications.

Herein, we report ultrasonic generation of thiyl radicals as a general method for functionalizing a range of surfaces with organic molecules. The method is simple, rapid, can be utilized at ambient conditions and involves sonicating a solution of disulfide molecules, homolytically cleaving S-S bonds and generating thiyl radicals that react with the surfaces by forming covalently bound monolayers. Full molecular coverages on conducting oxides (ITO), semiconductors (Si-H), and carbon (GC) electrode surfaces can be achieved within a time scale of 15-90 min. The suitability of this method to connect the same molecule to different electrodes enabled comparing the conductivity of single molecules and the electrochemical electron transfer kinetics of redox active monolayers as a function of the molecule-electrode contact. We demonstrate, using STM break-junction technique, single-molecule heterojunction comprising Au-molecule-ITO and Au-molecule-carbon circuits. We found that despite using the same molecule, the single-molecule conductivity of Au-molecule-carbon circuits is about an order of magnitude higher than that of Au-molecule-ITO circuits. The same trend was observed for electron transfer kinetics, measured using electrochemical impedance spectroscopy for ferrocene-terminated monolayers on carbon and ITO. This suggests that the interfacial bond between different electrodes and the same molecule can be used to tune the conductivity of single-molecule devices and to control the rate of charge transport in redox active monolayers, opening prospects for relating various types of interfacial charge-transfer rate processes.

Covalent Linkages of Molecules and Proteins to Si-H Surfaces Formed by Disulfide Reduction.

Thiols and disulfide contacts have been, for decades, key for connecting organic molecules to surfaces and nanoclusters as they form self-assembled monolayers (SAMs) on metals such as gold (Au) under mild conditions. In contrast, they have not been similarly deployed on Si owing to the harsh conditions required for monolayer formation. Here, we show that SAMs can be simply formed by dipping Si-H surfaces into dilute solutions of organic molecules or proteins comprising disulfide bonds. We demonstrate that S-S bonds can be spontaneously reduced on Si-H, forming covalent Si-S bonds in the presence of traces of water, and that this grafting can be catalyzed by electrochemical potential. Cyclic disulfide can be spontaneously reduced to form complete monolayers in 1 h, and the reduction can be catalyzed electrochemically to form full surface coverages within 15 min. In contrast, the kinetics of SAM formation of the cyclic disulfide molecule on Au was found to be three-fold slower than that on Si. It is also demonstrated that dilute thiol solutions can form monolayers on Si-H following oxidation to disulfides under ambient conditions; the supply of too much oxygen, however, inhibits SAM formation. The electron transfer kinetics of the Si-S-enabled SAMs on Si-H is comparable to that on Au, suggesting that Si-S contacts are electrically transmissive. We further demonstrate the prospect of this spontaneous disulfide reduction by forming a monolayer of protein azurin on a Si-H surface within 1 h. The direct reduction of disulfides on Si electrodes presents new capabilities for a range of fields, including molecular electronics, for which highly conducting SAM-electrode contacts are necessary and for emerging fields such as biomolecular electronics as disulfide linkages could be exploited to wire proteins between Si electrodes, within the context of the current Si-based technologies.

Reduced graphene oxide-silicon interface involving direct Si-O bonding as a conductive and mechanical stable ohmic contact.

Metal-semiconductor junctions are essential contacts for semiconductor devices, but high contact junction resistance is a limiting operational factor. Here, we establish an ohmic contact of low resistance of <4 × 10-6 Ω cm2 between platinum and n-type Si (111)-H surfaces. This involved Si-O covalent bonding a monolayer of graphene oxide (GO) to the Si surface followed by electrochemical reduction to form reduced graphene oxide (rGO). Current-voltage plots demonstrate that the GO/rGO transformation is associated with a change from a rectifying to an ohmic contact. The process is a viable method for constructing semiconductor-rGO interfaces and demonstrates that GO/rGO monolayers can be used as active components in tuning the contact resistance of metal-semiconductor junctions.

Spontaneous S-Si bonding of alkanethiols to Si(111)-H: towards Si-molecule-Si circuits.

We report the synthesis of covalently linked self-assembled monolayers (SAMs) on silicon surfaces, using mild conditions, in a way that is compatible with silicon-electronics fabrication technologies. In molecular electronics, SAMs of functional molecules tethered to gold via sulfur linkages dominate, but these devices are not robust in design and not amenable to scalable manufacture. Whereas covalent bonding to silicon has long been recognized as an attractive alternative, only formation processes involving high temperature and/or pressure, strong chemicals, or irradiation are known. To make molecular devices on silicon under mild conditions with properties reminiscent of Au-S ones, we exploit the susceptibility of thiols to oxidation by dissolved O2, initiating free-radical polymerization mechanisms without causing oxidative damage to the surface. Without thiols present, dissolved O2 would normally oxidize the silicon and hence reaction conditions such as these have been strenuously avoided in the past. The surface coverage on Si(111)-H is measured to be very high, 75% of a full monolayer, with density-functional theory calculations used to profile spontaneous reaction mechanisms. The impact of the Si-S chemistry in single-molecule electronics is demonstrated using STM-junction approaches by forming Si-hexanedithiol-Si junctions. Si-S contacts result in single-molecule wires that are mechanically stable, with an average lifetime at room temperature of 2.7 s, which is five folds higher than that reported for conventional molecular junctions formed between gold electrodes. The enhanced "ON" lifetime of this single-molecule circuit enables previously inaccessible electrical measurements on single molecules.

Condensin Smc2-Smc4 Dimers Are Flexible and Dynamic.

Structural maintenance of chromosomes (SMC) protein complexes, including cohesin and condensin, play key roles in the regulation of higher-order chromosome organization. Even though SMC proteins are thought to mechanistically determine the function of the complexes, their native conformations and dynamics have remained unclear. Here, we probe the topology of Smc2-Smc4 dimers of the S. cerevisiae condensin complex with high-speed atomic force microscopy (AFM) in liquid. We show that the Smc2-Smc4 coiled coils are highly flexible polymers with a persistence length of only ∼ 4 nm. Moreover, we demonstrate that the SMC dimers can adopt various architectures that interconvert dynamically over time, and we find that the SMC head domains engage not only with each other, but also with the hinge domain situated at the other end of the ∼ 45-nm-long coiled coil. Our findings reveal structural properties that provide insights into the molecular mechanics of condensin complexes.