Specifically horizontally tethered DNA probes on Au surfaces allow labelled and label-free DNA detection using SERS and electrochemically driven melting.

Controlled covalent attachment of dsDNA horizontally orientated on a gold surface is achieved through the use of a single surface-linker located approximately half way along the attached DNA probe strand. We show that horizontally oriented dsDNA on a gold surface can undergo melting and re-hybridization to target strand in solution and thus can be used for the detection of specific target DNA sequences using surface-enhanced Raman spectroscopy (SERS). We show that a range of lengths of target DNA sequences from ∼30-bases to 78-bases can be specifically hybridized to the short immobilized DNA probe sequence and adopt a horizontal orientation on the gold surface. Following thermal or electrochemically driven melting of the immobilized dsDNA, the target DNA strand diffuses away while the probe strand remains attached to the surface allowing the functionalized surfaces to be reused. The melting of the horizontally orientated immobilized dsDNA can be monitored using SERS either by employing a dye label covalently attached on the DNA target strand or by employing a binding agent selective for dsDNA. This approach of covalently immobilizing the DNA probe strand through a linker located at approximately the middle of the strand has great potential to improve the sensitivity and specificity of molecular assays that employ DNA arrays on solid surfaces.

Strain discrimination of Yersinia pestis using a SERS-based electrochemically driven melting curve analysis of variable number tandem repeat sequences.

Strain discrimination within genetically highly similar bacteria is critical for epidemiological studies and forensic applications. An electrochemically driven melting curve analysis monitored by SERS has been utilised to reliably discriminate strains of the bacterial pathogen Yersinia pestis, the causative agent of plague. DNA amplicons containing Variable Number Tandem Repeats (VNTRs) were generated from three strains of Y. pestis: CO92, Harbin 35 and Kim. These amplicons contained a 10 base pair VNTR repeated 6, 5, and 4 times in CO92, Harbin 35 and Kim respectively. The assay also included a blocker oligonucleotide comprising 3 repeats of the 10-mer VNTR sequence. The use of the blocker reduced the effective length of the target sequence available to bind to the surface bound probe and significantly improved the sensitivity of the discrimination. The results were consistent during three replicates that were carried out on different days, using different batches of PCR product and different SERS sphere segment void (SSV) substrate. This methodology which combines low cost, speed and sensitivity is a promising alternative to the time consuming current electrophoretic methods.

Electrodeposition from supercritical fluids.

Recent studies have shown that it is possible to electrodeposit a range of materials, such as Cu, Ag and Ge, from various supercritical fluids, including hydrofluorocarbons and mixtures of CO2 with suitable co-solvents. In this perspective we discuss the relatively new field of electrodeposition from supercritical fluids. The perspective focuses on some of the underlying physical chemistry and covers both practical and scientific aspects of electrodeposition from supercritical fluids. We also discuss possible applications for supercritical fluid electrodeposition and suggest some key developments that are required to take the field to the next stage.

Influence of macroporous gold support and its functionalization on lactate oxidase-based biosensors response.

A general bioanalytical platform for biosensor applications was developed based on three-dimensional ordered macroporous (3DOM) gold film modified electrodes using lactate oxidase (LOx) as a case study, within the framework of developing approaches of broad applicability. The electrode was electrochemically fabricated with an inverted opal template, making the surface area of the 3DOM gold electrode up to 18 times higher than that of bare flat gold electrodes. These new electrochemical transducers were characterized by using Field Emission Scanning Electron Microscopy (FE-SEM), Atomic Force Microscopy (AFM) and the X-ray diffraction (XRD). The biosensor was developed by immobilization of lactate oxidase (LOx), on a 3DOM gold electrode modified with a self-assembled monolayer of dithiobis-N-succinimidyl propionate (DTSP). The resulting lactate oxidase biosensor was characterized by electrochemical impedance spectroscopy (EIS). The 3DOM gold electrode not only provides a good biocompatible microenvironment but also promotes the increase of conductivity and stability. Thus, the developed lactate oxidase bioanalytical platforms showed higher mediated bioelectrocatalytic activity compared to others previously described based on polycrystalline gold transducers. The response to varying lactate concentrations has been obtained in the presence of hydroxymethylferrocene as redox mediator in solution. Under these conditions, the bioanalytical platform response for DTSP covalently bound enzyme was improved with respect to that obtained in absence of DTSP.

Enhancing solar cells with localized plasmons in nanovoids.

Localized plasmon resonances of spherical nanovoid arrays strongly enhance solar cell performance by a factor of 3.5 in external quantum efficiency at plasmonic resonances, and a four-fold enhancement in overall power conversion efficiency. Large area substrates of silver nanovoids are electrochemically templated through self-assembled colloidal spheres and organic solar cells fabricated on top. Our design represents a new class of plasmonic photovoltaic enhancement: that of localized plasmon-enhanced absorption within nanovoid structures. Angularly-resolved spectra demonstrate strong localized Mie plasmon modes within the nanovoids. Theoretical modelling shows varied spatial dependence of light intensity within the void region suggesting a first possible route towards Third Generation plasmonic photovoltaics.

An analysis of the kinetics of oxidation of ascorbate at poly(aniline)-poly(styrene sulfonate) modified microelectrodes.

A detailed kinetic study is provided for the oxidation of ascorbate at poly(aniline)-poly(styrene sulfonate) coated microelectrodes. Flat films with a low degree of polymer spillover and a thickness much lower than the microelectrode radius were produced by controlled potentiodynamic electrodeposition. The currents for ascorbate oxidation are found to be independent of the polymer thickness, indicating that the reaction occurs at the outer surface of the polymer film. At low ascorbate concentrations, below around 40 mM, the currents are found to be mass transport limited. At higher ascorbate concentrations the currents became kinetically limited. The experimental data for measurements at a range of potentials are fitted to a consistent kinetic model and the results summarized in a case diagram. The results obtained for the poly(aniline)-(polystyrene sulfonate) coated microelectrode are compared to those for a poly(aniline)-poly(vinyl sulfonate) coated microelectrode and to the results of an earlier study of the reaction at poly(aniline)-poly(vinyl sulfonate) coated rotating disc electrodes. For poly(aniline)-poly(styrene sulfonate) the oxidation of ascorbate is found to proceed by one electron reaction whereas for poly(aniline)-poly(vinyl sulfonate) the reaction is found to be a two electron oxidation.

The oxidation of ascorbate at copolymeric sulfonated poly(aniline) coated on glassy carbon electrodes.

Self-doped poly(aniline)s as electrode coatings to catalyze ascorbate oxidation are revisited in this article. Sulfonated poly(aniline) (SPAN) was deposited on glassy carbon electrodes as a copolymer of aniline and its sulfonated derivative, 2-aminobenzenesulfonic acid (2-ABSA). The resulting deposits are reproducible and show good stability and electroactivity at pH>7, enabling studies at typical physiological pH values. Calibration curves were obtained using a rotating disc electrode at a sampling potential of 0.2V, displaying linear dependence in the region 0-20mM ascorbate. A kinetic model based on the Michaelis-Menten reaction mechanism, previously validated for poly(aniline) composites, was used to analyse the form of the calibration curve leading to values of the effective reaction constants K(ME) and k'(ME). New calibration curves constructed for different sampling potentials were used to elucidate the rate limiting step at saturated kinetics. Rotating disc voltammetry performed at increasing pH (from pH 2 to 9) showed a dramatic decrease in the limiting current, without any evidence for a change in the reaction mechanism.

Electrodeposition of PANi films on platinum needle type microelectrodes. Application to the oxidation of ascorbate in human plasma.

Needle type (165 microm(2)) and small Pt disc (3-11 microm(2)) microelectrodes were used for the electrodeposition of composite poly(aniline), PANi, films by cyclic voltammetry and chronoamperometry for the oxidation of ascorbate. PANi electroactivity at neutral pH was retained through polymer alkylation or by using large poly-anions, such as poly(vinylsulfonate), PVS, and poly(styrenesulfonate), PSS. Hence the growth of the composite films was studied in the presence of different counter ions such as SO(4)(2-), Cl(-), NaPVS and NaPSS. The morphology of the resulting films was examined by scanning electron microscopy. Results showed that flat PANi films with thicknesses much lower than the microelectrode radius were obtained by potentiodynamic electrodeposition. On the other hand films with mushroom shapes, with significant spill over, were obtained under constant potential. The resulting polymer modified microelectrodes films were shown to be suitable for the oxidation of ascorbate at 0.1 V vs. SCE and pH 7 with a detection limit of 1 microM for PANi/PSS composites. The current was independent of film thickness, mass transport controlled at low ascorbate concentrations and not affected by the presence of common interferences such as uric acid, glutathione or vitamin E. Due to their excellent properties the PANi-PSS film coated microelectrodes were used for the amperometric detection of ascorbate in human plasma. The results are encouraging for the use of small polymer modified Pt needle type microelectrodes for the detection of ascorbate in biological systems.

The fabrication of plasmonic Au nanovoid trench arrays by guided self-assembly.

Metallic nanoscale voids can support both localized and propagating plasmons and form plasmonic crystals. However, constructing 1D arrays is crucial for producing plasmonic circuits. In this paper we report the first experimental evidence of plasmons in templated linear arrays of self-assembled structures. Single and multilayer arrays of gold voids have been fabricated by self-assembly of sub-micron polystyrene spheres in V-shaped trenches in silicon, followed by selective area electrodeposition. Angle-dependent dispersion characteristics reveal the existence of localized plasmons.

UV SERS at well ordered Pd sphere segment void (SSV) nanostructures.

Ultraviolet laser excited surface-enhanced Raman scattering was obtained for the first time at the well ordered palladium sphere segment void (SSV) nanostructures, using adenine as the probe molecule, and the UV-SERS enhancement is found to be correlated well with the plasmon absorption of Pd SSVs in the UV region.

Extracting kinetic parameters for homogeneous [Os(bpy)2ClPyCOOH]+ mediated enzyme reactions from cyclic voltammetry and simulations.

The homogeneous reaction between glucose oxidase and osmium bipyridine-pyridine carboxylic acid in the presence of glucose has been studied in detail by cyclic voltammetry and digital simulation. Combination of the analytical equations that describe the dependence of the amperometric response on enzyme, substrate and co-substrate concentrations for the limiting cases with digital simulation of the coupled enzyme reaction diffusion problem allows us to extract kinetic parameters for the substrate-enzyme reaction: K(MS)=10.8 mM, k(cat)=254 s(-1) and for the redox mediator-enzyme reaction, k=2.2x10(5) M(-1) s(-1). The accurate determination of the kinetic parameters at low substrate concentrations (<7 mM) is limited by depletion of the substrate close to the electrode surface. At high substrate concentrations (>20 mM) inactivation of the reduced form of glucose oxidase in the bulk solution must be taken into account in the analysis of the results.

Relaxation and Simplex mathematical algorithms applied to the study of steady-state electrochemical responses of immobilized enzyme biosensors: Comparison with experiments.

A description of the implementation of the relaxation method with automatic mesh point allocation for immobilized enzyme electrodes is presented. The advantages of this method for the solution of coupled reaction-diffusion problems are discussed. The relaxation numerical simulation technique is combined with the Simplex fitting algorithm to extract kinetic parameters from experimental data. The results of the simulations are compared to experimental data from self-assembled multilayered electrodes comprised of glucose oxidase (GOx) and an Os modified redox mediator and found to be in excellent agreement.

Easily coupled whispering gallery plasmons in dielectric nanospheres embedded in gold films.

A new self-aligned robust method for coupling to whispering gallery modes (WGMs) of submicron microspheres utilizes their periodic arrangement without relying on nanopositioned external coupling devices. The microspheres are embedded in a nanostructured gold surface supporting delocalized plasmonic crystal modes that mediate the coupling, and can be tuned by the geometry. Detailed measurements of the angle- and orientation-dependent reflectivity reveal localized plasmonic WGMs whose energies scale with sphere diameter and agree closely with Mie calculations. Coupling between these plasmonic WGMs leads to mode splitting and the formation of plasmonic minibands of a controllable bandwidth.

Strong coupling between localized plasmons and organic excitons in metal nanovoids.

Hybrid emitting exciton-plasmonic composites are constructed by coating arrays of spherical nanovoids embedded in a gold film with organic semiconducting molecular J-aggregate films. In such plasmonic crystals, localized plasmons confined inside the voids can be excited. We report the first observation of polaritonic spectral narrowing and strong coupling between localized plasmons and J-aggregate excitons with Rabi splittings of 230 meV at room temperature.

Plasmonic band gaps and trapped plasmons on nanostructured metal surfaces.

Nanostructured metal surfaces comprised of periodically arranged spherical voids are grown by electrochemical deposition through a self-assembled template. Detailed measurements of the angle- and orientation-dependent reflectivity reveal the spectral dispersion, from which we identify the presence of both delocalized Bragg and localized Mie plasmons. These couple strongly producing bonding and antibonding mixed plasmons with anomalous dispersion properties. Appropriate plasmon engineering of the void morphology selects the plasmon spatial and spectral positions, allowing these plasmonic crystal films to be optimized for a wide range of sensing applications.

Voltammetry and determination of metronidazole at a carbon fiber microdisk electrode.

The electrochemistry of metronidazole, 1-(hydroxyethyl)-2-methyl-5-nitroimidazole, was investigated at a carbon fiber microdisk electrode in pH 9 Britton Robinson buffer. Under these conditions, the reduction of metronidazole is controlled by both mass transport to the microdisk and adsorption with an equilibrium constant of 4x10(3)mol(-1)dm(3) and a saturation coverage of 0.88x10(-8)molcm(-2). The adsorption and accumulation of metronidazole on the surface of the carbon fiber allows its determination at low concentrations by square wave adsorptive stripping voltammetry. A detection limit for metronidazole of 5x10(-7)moldm(-3) and a R.S.D. of 3.7% at 1x10(-6)moldm(-3) (n=4) were obtained with a two electrode system with no stirring during the accumulation step. Based on this method, a simple procedure for the determination of metronidazole in urine is described which requires no pre-treatment of the sample before analysis.

Tunable resonant optical microcavities by self-assembled templating.

Micrometer-scale optical cavities are produced by a combination of template sphere self-assembly and electrochemical growth. Transmission measurements of the tunable microcavities show sharp resonant modes with Q factors of >300 and 25-fold local enhancement of light intensity. The presence of transverse optical modes confirms the lateral confinement of photons. Calculations show that submicrometer mode volumes are feasible. The small mode volumes of these microcavities promise to lead to a wide range of applications. in microlasers, atom optics, quantum information, biophotonics, and single-molecule detection.

The demonstration of an enhanced microelectrochemical transistor for measurements in neutral solution at low analyte concentration.

We report a new, sensitive detection method for the measurement of analytes based on the combination of conducting polymer technology and redox enzyme electrochemistry. This method provides a simple measurement method potentially enabling point of care testing on disposable electrodes in under 60 s. The measurement does not require a reference or counter electrode and the analytical signal is recorded by an ammeter measuring milliampere currents through a polymer transistor. Even at very low analyte concentration, the signal is not subject to significant noise.

The measurement of alkaline phosphatase at nanomolar concentration within 70 s using a disposable microelectrochemical transistor.

We report a new approach to the measurement of alkaline phosphatase concentration based on the use of a disposable poly(aniline) microelectrochemical transistor. The measurement is carried out in a two cell configuration in which the poly(aniline) microelectrochemical transistor operates in acid solution and is connected to the alkaline buffer solution containing the analyte by a salt bridge. Disposable microelectrochemical transistors were reproducibly fabricated by electrochemical deposition of poly(aniline) onto photolithographically fabricated gold microband arrays. Using these devices alkaline phosphatase was detected by employing p-aminophenyl phosphate as the substrate for the enzyme and using glucose and glucose oxidase to recycle the p-aminophenol generated upon enzyme catalysed hydrolysis of the phosphate. Recycling the p-aminophenol with glucose and glucose oxidase amplified the detection of alkaline phosphatase approximately tenfold. Using this approach we obtain linear calibration curves for alkaline phosphatase up to 5 nM within 70 s on single use devices.

Optical properties of nanostructured metal films.

Nanostructured metal films of platinum, gold and silver up to 675 nm thick we prepared by electrochemical deposition through templates of 700 nm diameter polystyrene spheres assembled as hexagonal close packed monolayer on an evaporated gold surface followed by removal of the template by dissolution in tetrahydrofuran. The reflection spectra of the films at normal incidence were recorded as a function of film thickness and the spectra correlated with the local visual appearance of the film and the surface structure from SEM. For thin films, below one quarter sphere height, the spectra show a single reflectivity dip at a wavelength just below the sphere diameter consistent with surface-plasmon grating-like behaviour. For the thicker films several reflectivity dips are observed which move towards longer wavelength with increasing film thickness. This behaviour is shown to be consistent with a model in which light reflected from the top of the structure interferes with light reflected from within the spherical segment cavities in the film.