Electrochemistry at the Edge of a van der Waals Heterostructure.

Artificial van der Waals heterostructures, obtained by stacking two-dimensional (2D) materials, represent a novel platform for investigating physicochemical phenomena and applications. Here, the electrochemistry at the one-dimensional (1D) edge of a graphene sheet, sandwiched between two hexagonal boron nitride (hBN) flakes, is reported. When such an hBN/graphene/hBN heterostructure is immersed in a solution, the basal plane of graphene is encapsulated by hBN, and the graphene edge is exclusively available in the solution. This forms an electrochemical nanoelectrode, enabling the investigation of electron transfer using several redox probes, e.g., ferrocene(di)methanol, hexaammineruthenium, methylene blue, dopamine and ferrocyanide. The low capacitance of the van der Waals edge electrode facilitates cyclic voltammetry at very high scan rates (up to 1000 V s-1), allowing voltammetric detection of redox species down to micromolar concentrations with sub-second time resolution. The nanoband nature of the edge electrode allows operation in water without added electrolyte. Finally, two adjacent edge electrodes are realized in a redox-cycling format. All the above-mentioned phenomena can be investigated at the edge, demonstrating that nanoscale electrochemistry is a new application avenue for van der Waals heterostructures. Such an edge electrode will be useful for studying electron transfer mechanisms and the detection of analyte species in ultralow sample volumes.

Graphene-on-gold surface plasmon resonance sensors resilient to high-temperature annealing.

Gold films coated with a graphene sheet are being widely used as sensors for the detection of label-free binding interactions using surface plasmon resonance (SPR). During the preparation of such sensors, it is often essential to subject the sensor chips to a high-temperature treatment in order to ensure a clean graphene surface. However, sensor chips used currently, which often use chromium as an adhesion promoter, cannot be subjected to temperatures above 250 °C, because under such conditions, chromium is found to reorganize and diffuse to the surface, where it is easily oxidized, impairing the quality of SPR spectra. Here we present an optimized preparation strategy involving a three-cycle tempering coupled with chromium (oxide) etching, which allows the graphene-coated SPR chips to be annealed up to 500 °C with little deterioration of the surface morphology. In addition, the treatment delivers a surface that shows a clear enhancement in spectral response together with a good refractive index sensitivity. We demonstrate the applicability of our sensors by studying the kinetics of avidin-biotin binding at different pH repeatedly on the same chip. The possibility to anneal can be exploited to recover the original surface after sensing trials, which allowed us to reuse the sensor for at least six cycles of biomolecule adsorption.

Statistical Copolymers that Mimic Aspects of Mussel Adhesive Proteins: Access to Robust Adhesive-Domains for Non-Covalent Surface PEGylation.

Reconstructing functional sequence motifs of proteins, using statistical copolymers greatly reduces the information content, but simplifies synthesis significantly. Key amino acid residues involved in the adhesion of mussel foot proteins are identified. The side-chain functionalities of Dopa, lysine, and arginine are abstracted and incorporated into acrylate monomers to allow controlled radical polymerization. The resulting Dopa-acrylate (Y*-acr), arginine-acrylate (R-acr), and lysine-acrylate (K-acr) monomers are polymerized in different monomer ratios and compositions by reversible addition fragmentation transfer polymerization with a poly(ethylene glycol) (PEG) macrochain transfer agent. This results in two sets of PEG-block-copolymers with statistical mixtures and different monomer ratios of catechol/primary amine and catechol/guanidine side-chain functionalities, both important pairs for mimicking π-cation interactions. The coating behavior of these PEG-block-copolymers is evaluated using quartz crystal microbalance with dissipation energy monitoring (QCM-D), leading to non-covalent PEGylation of the substrates with clear compositional optima in the coating stability and antifouling properties. The coatings prevent non-reversible albumin or serum adsorption, as well as reduce cellular adhesion and fungal spore attachment.

Photoactive Yellow Protein Adsorption at Hydrated Polyethyleneimine and Poly-l-Glutamic Acid Interfaces.

Chiral and achiral vibrational sum-frequency generation (VSFG) spectroscopy was performed in the 1400-1700 and 2800-3800 cm-1 range to study the interfacial structure of photoactive yellow protein (PYP) adsorbed on polyethyleneimine (PEI) and poly-l-glutamic acid (PGA) surfaces. Nanometer-thick polyelectrolyte layers served as the substrate for PYP adsorption, with 6.5-pair layers providing the most homogeneous surfaces. When the topmost material was PGA, it acquired a random coil structure with a small number of ß2-fibrils. Upon adsorption on oppositely charged surfaces, PYP yielded similar achiral spectra. However, the VSFG signal intensity increased for PGA surfaces with a concomitant redshift of the chiral Cα-H and N-H stretching bands, suggesting increased adsorption for PGA compared to PEI. At low wavenumbers, both the backbone and the side chains of PYP induced drastic changes to all measured chiral and achiral VSFG spectra. Decreasing ambient humidity led to the loss of tertiary structure with a re-orientation of α-helixes, evidenced by a strongly blue-shifted chiral amide I band of the ß-sheet structure with a shoulder at 1654 cm-1. Our observations indicate that chiral VSFG spectroscopy is not only capable of determining the main type of secondary structure of PYP, i.e., ß-scaffold, but is also sensitive to tertiary protein structure.

Faradaic effects in electrochemically gated graphene sensors in the presence of redox active molecules.

Field-effect transistors (FETs) based on graphene are promising devices for the direct sensing of a range of analytes in solution. We show here that the presence of redox active molecules in the analyte solution leads to the occurrence of heterogeneous electron transfer with graphene generating a Faradaic current (electron transfer) in a FET configuration resulting in shifts of the Dirac point. Such a shift occurs if the Faradaic current is significantly high, e.g. due to a large graphene area. Furthermore, the redox shift based on the Faradaic current, reminiscent of a doping-like effect, is found to be non-Nernstian and dependent on parameters known from electrode kinetics in potentiodynamic methods, such as the electrode area, the standard potential of the redox probes and the scan rate of the gate voltage modulation. This behavior clearly differentiates this effect from other transduction mechanisms based on electrostatic interactions or molecular charge transfer doping effects, which are usually behind a shift of the Dirac point. These observations suggest that large-area unmodified/pristine graphene in field-effect sensors behaves as a non-polarized electrode in liquid. Strategies for ensuring a polarized interface are discussed.

A primary battery-on-a-chip using monolayer graphene.

We present here a bottom-up approach for realizing on-chip on-demand batteries starting out with chemical vapor deposition-grown graphene. Single graphene monolayers contacted by electrode lines on a silicon chip serve as electrodes. The anode and cathode are realized by electrodeposition of zinc and copper respectively onto graphene, leading to the realization of a miniature graphene-based Daniell cell on a chip. The electrolyte is housed partly in a gel and partly in liquid form in an on-chip enclosure molded using a 3d printer or made out of poly(dimethylsiloxane). The realized batteries provide a stable voltage (∼1.1 V) for many hours and exhibit capacities as high as 15 µAh, providing enough power to operate a pocket calculator. The realized batteries show promise for deployment as on-chip power sources for autonomous systems in lab-on-a-chip or biomedical applications.

Rolling circle amplification-based detection of human topoisomerase I activity on magnetic beads.

A high-sensitivity assay has been developed for the detection of human topoisomerase I with single molecule resolution. The method uses magnetic sepharose beads to concentrate rolling circle products, produced by the amplification of DNA molecules circularized by topoisomerase I and detectable with a confocal microscope as single and discrete dots, once reacted with fluorescent probes. Each dot, corresponding to a single cleavage-religation event mediated by the enzyme, can be counted due to its high signal/noise ratio, allowing detection of 0.3pM enzyme and representing a valid method to detect the enzyme activity in highly diluted samples.

Real-Time Monitoring of Cell Adhesion onto a Soft Substrate by a Graphene Impedance Biosensor.

Soft substrates are interesting for many applications, ranging from mimicking the cellular microenvironment to implants. Conductive electrodes on such substrates allow the realization of flexible, elastic, and transparent sensors. Single-layer graphene as a candidate for such electrodes brings the advantage that the active area of the sensor is transparent and conformal to the underlying substrate. Here, we overcome several challenges facing the routine realization of graphene cell sensors on a canonical soft substrate, namely, poly(dimethylsiloxane) (PDMS). We have systematically studied the effect of surface energy before, during, and after the transfer of graphene. Thus, we have identified a suitable support polymer, optimal substrate (pre)treatment, and an appropriate solvent for the removal of the support. Using this procedure, we can reproducibly obtain stable and intact graphene sensors on a millimeter scale on PDMS, which can withstand continuous measurements in cell culture media for several days. From local nanomechanical measurements, we infer that the softness of the substrate is slightly affected after the graphene transfer. However, we can modulate the stiffness using PDMS with differing compositions. Finally, we show that graphene sensors on PDMS can be successfully used as soft electrodes for real-time monitoring of the cell adhesion kinetics. The routine availability of single-layer graphene electrodes on a soft substrate with tunable stiffness will open a new avenue for studies, where the PDMS-liquid interface is made conducting with minimal alteration of the intrinsic material properties such as softness, flexibility, elasticity, and transparency.

Towards in vitro molecular diagnostics using nanostructures.

Nanostructures appear to be promising for a number of applications in molecular diagnostics, mainly due to the increased surface-to-volume ratio they can offer, the very low limit of detection achievable, and the possibility to fabricate point-of-care diagnostic devices. In this paper, we review examples of the use of nanostructures as diagnostic tools that bring in marked improvements over prevalent classical assays. The focus is laid on the various sensing paradigms that possess the potential or have demonstrated the capability to replace or augment current analytical strategies. We start with a brief introduction of the various types of nanostructures and their physical properties that determine the transduction principle. This is followed by a concise collection of various functionalization protocols used to immobilize biomolecules on the nanostructure surface. The sensing paradigms are discussed in two contexts: the nanostructure acting as a label for detection, or the nanostructure acting as a support upon which the molecular recognition events take place. In order to be successful in the field of molecular diagnostics, it is important that the nanoanalytical tools be evaluated in the appropriate biological environment. The final section of the review compiles such examples, where the nanostructure-based diagnostic tools have been tested on realistic samples such as serum, demonstrating their analytical power even in the presence of complex matrix effects. The ability of nanodiagnostic tools to detect ultralow concentrations of one or more analytes coupled with portability and the use of low sample volumes is expected to have a broad impact in the field of molecular diagnostics.

A highly durable graphene monolayer electrode under long-term hydrogen evolution cycling.

Achieving long term stability of single graphene sheets towards repeated electrochemical hydrogen evolution reaction (HER) cycling has been challenging. Here, we show through appropriate electrode preparation that it is possible to obtain highly durable isolated graphene electrodes, which can survive several hundreds of HER cycles with virtually no damage to the sp2-carbon framework and persistently good electron transfer characteristics.

Enzyme-free sugar sensing in microfluidic channels with an affinity-based single-wall carbon nanotube sensor.

We present a novel nonenzymatic carbon nanotube sensor integrated in a microfluidic channel for the detection of sugars. The sensor is assembled as a liquid-gated field-effect transistor, with the transistor channel composed of 1 to 10 nanotubes, which are controllably functionalized with boronic acid receptors. The devices show sensitivity to glucose in a concentration range of 5 to 30 mM. Furthermore, by controlling the type of nanotube-receptor coupling (as covalent or noncovalent) and by deploying a sensitive impedance-based detection technique, we corroborate in detail the transduction mechanism of our affinity-based sensor. In the case of covalent coupling, charge carrier scattering along the nanotubes is the dominant mechanism. While in the noncovalent case, surface charge effects dominate. The identification of the mechanism along with the tunability of the chemical coupling and the cost-effective integration in microchannels constitute a solid basis for the entry of nanotube-based sensors in lab-on-a-chip applications.

Site-specific self-assembled liquid-gated ZnO nanowire transistors for sensing applications.

A scalable bottom-up solution-based approach for the site-specific realization of ZnO nanowire (ZnO-NW)-based field-effect transistors for sensing applications in liquids is reported. The nanowires are grown across predefined electrodes patterned by photolithography. Site specificity is attained by the use of nanoparticles acting as seeds. Using integrated on-chip microchannels and microfabricated gate electrodes, electrochemically gated ZnO-NW network transistors functioning in liquids are demonstrated. The optimized devices are rendered sensitive to pH through chemical functionalization. The unique combination of the sensitivity, site specificity, scalability, and cost effectiveness of the technique opens up avenues for the routine realization of one-dimensional nanostructure-based chemical and biosensors for analytical and diagnostic applications.

Marker-free on-the-fly fabrication of graphene devices based on fluorescence quenching.

Graphene has been dominating the electronic research community recently, with a brisk surge in proposals for its use in novel devices. The aspirations of 2D-carbon-based electronics largely rely on the availability of a mass-production technique to obtain wafer-scale graphene circuits. In this paper, we take a first step towards fulfilling this aspiration by demonstrating a rapid prototyping route for graphene-based devices. The method is based on our observation that graphene quenches the fluorescence from dyes. Utilizing this property, we use a confocal microscope to identify graphene flakes and perform the required lithography steps, bypassing the need for markers and other infrastructure such as atomic force microscopy or e-beam lithography. The versatility of this technique enables it to harbour ambitions of an automated process for large scale in situ assembly of graphene-based circuits.

Effect of stacking order on the electric-field induced carrier modulation in graphene bilayers.

When planar graphene sheets are stacked on top of each other, the electronic structure of the system varies with the position of the subsequent sublattice atoms. Here, we employ scanning photocurrent microscopy to study the disparity in the behavior of charge carriers for two different stacking configurations. It has been found that deviation from the regular Bernal stacking decouples the sheets from each other, which imparts effective electrostatic screening of the farther layer from the underlying backgate. Electrochemical top-gating is demonstrated as a means to selectively tune the charge carrier density in the decoupled upper layer.

Vertical arrays of nanofluidic channels fabricated without nanolithography.

Vertical arrays of sealed nanofluidic channels, in which both cross-sectional dimensions are controllable down to 10 nm, were fabricated by selective side etching of a SiGe heterostructure comprised of layers of alternating Ge fractions. Capillary filling of these nanochannel arrays with fluorescent dye solutions was investigated using a confocal microscope. The feasibility of using nanochannels for size-based separation of biomolecules was demonstrated by imaging aggregates of tagged amyloid-beta peptide. The ability to integrate a large number of nanochannels shows promise for high throughput applications involving lab-on-a-chip systems.

Selective enhancement of carbon nanotube photoluminescence by resonant energy transfer.

Highly selective: Enhancement of the photoluminescence (PL) emission efficiency of selected chiral forms of semiconducting single-walled carbon nanotubes (SWCNTs) is presented (see figure). Excitation of Nile blue A in the presence of SWCNTs results in the quenching of its fluorescence. The energy is resonantly transferred to the (7,5) SWCNT whereas the (8,7) tube is not in resonance; hence, its PL remains unaffected.We report on a simple method for enhancing the efficiency of photoluminescence (PL) emission from selected chiral forms of semiconducting single-wall carbon nanotubes (SWCNTs). The method is based on the use of a fluorescent dye (Nile blue A) that shows the capability of resonant energy transfer on to nanotubes. The excitation of Nile blue A in the presence of SWCNTs results in the quenching of its fluorescence and the energy is resonantly transferred to certain chiral forms. The PL emission from these chiral forms shows a marked increase in efficiency signifying the occurrence of Förster type resonant energy transfer (FRET). Due to its simplicity, this procedure has widespread implications for the detection of carbon nanotubes as well as for their use as fluorophores in FRET-based in vivo and in vitro biological applications.

Applications of the static quenching of rhodamine B by carbon nanotubes.

We report here on the interaction of the fluorescent dye rhodamine B (RB) with single-walled carbon nanotubes (SWCNTs). We observe that SWCNTs statically quench the fluorescence of RB by forming a stable ground state complex. Careful spectroscopic analysis indicates that the complex formation is efficient mainly with certain chiral forms. We propose three different applications utilizing this quenching mechanism and the associated complexation. Firstly, the quenching efficiency can be utilized as a measure for the characterization and quantification of nanotube dispersions. Secondly, we demonstrate that the specific complexation of RB can be deployed to enrich certain chiral forms in suspension. Finally, we show that RB can be effectively used to visualize nanotubes deposited on substrates.

Selective electrochemical functionalization of the graphene edge.

We present a versatile and simple method using electrochemistry for the exclusive functionalization of the edge of a graphene monolayer with metal nanoparticles or polymeric amino groups. The attachment of metal nanoparticles allows us to exploit surface-enhanced Raman scattering to characterize the chemistry of both the pristine and the functionalized graphene edge. For the pristine patterned graphene edge, we observe the typical edge-related modes, while for the functionalized graphene edge we identify the chemical structure of the functional layer by vibrational fingerprinting. The ability to obtain single selectively functionalized graphene edges routinely on an insulating substrate opens an avenue for exploring the effect of edge chemistry on graphene properties systematically.

pH sensitivity of interfacial electron transfer at a supported graphene monolayer.

Electrochemical devices based on a single graphene monolayer are often realized on a solid support such as silicon oxide, glassy carbon or a metal film. Here, we show that, with graphene on insulating substrates, the kinetics of the electron transfer at graphene with various redox active molecules is dictated by solution pH for electrode reactions that are not proton dependent. We attribute the origin of this unusual phenomenon mainly to electrostatic effects between dissolved/dissociated redox species and the interfacial charge due to trace amounts of ionizable groups at the supported graphene-liquid interface. Cationic redox species show higher electron transfer rates at basic pH, while anionic species undergo faster electron transfer at acidic pH. Although this behavior is observed on graphene on three different insulating substrates, the strength of this effect appears to differ depending on the surface charge density of the underlying substrate. This finding has important implications for the design of electrochemical sensors and electrocatalysts based on graphene monolayers.

Correction: pH sensitivity of interfacial electron transfer at a supported graphene monolayer.

Correction for 'pH sensitivity of interfacial electron transfer at a supported graphene monolayer' by Michel Wehrhold et al., Nanoscale, 2019, DOI: 10.1039/c9nr05049c.