P.R. Dunstan

Generic epitaxial graphene biosensors for ultrasensitive detection of cancer risk biomarker

A generic electrochemical method of ?bioreceptor? antibody attachment to phenyl amine functionalized graphitic surfaces is demonstrated. Micro-channels of chemically modified multi-layer epitaxial graphene (MLEG) have been used to provide a repeatable and reliable response to nano-molar (nM) concentrations of the cancer risk (oxidative stress) biomarker 8-hydroxydeoxyguanosine (8-OHdG). X-ray photoelectron spectroscopy, Raman spectroscopy are used to characterize the functionalized MLEG. Confocal fluorescence microscopy using fluorescent-labelled antibodies indicates that the anti-8-OHdG antibody selectively binds to the phenyl amine-functionalized MLEG?s channel. Current?voltage measurements on functionalized channels showed repeatable current responses from antibody?biomarker binding events. This technique is scalable, reliable, and capable of providing a rapid, quantitative, label-free assessment of biomarkers at nano-molar (<20 nM) concentrations in analyte solutions. The sensitivity of the sensor device was investigated using varying concentrations of 8-OHdG, with changes in the sensor?s channel resistance observed upon exposure to 8-OHdG. Detection of 8-OHdG concentrations as low as 0.1 ng ml?1 (0.35 nM) has been demonstrated. This is five times more sensitive than reported enzyme linked immunosorbent assay tests (0.5 ng ml?1).

Single-walled carbon nanotubes: differential genotoxic potential associated with physico-chemical properties

Abstract Single-walled carbon nanotubes (SWCNTs) have recently attracted great attention because of their fibrous structure and high aspect ratio. Here the genotoxic potential of 400–800 nm, 1–3 μm and 5–30 μm SWCNT with respect to their geometry and surface characteristics was studied. Following thorough physico-chemical characterisation, human bronchial epithelial (BEAS−2B) and lymphoblastoid (MCL-5) cells were treated with SWCNT for 24 or 48 h. This showed significant increases in micronucleus frequency in a time- and dose-dependent manner in both cell types in the absence of cytotoxicity. Over the same dose range, only 1–3 μm SWCNT gave rise to significant increases in hprt point mutations at doses ≥25 μg/ml. Cellular 2,7-dichlorodihydrofluoresceindiacetate (DCFH-DA) fluorescence assay and RT-PCR for oxidative pathway gene profiling revealed a possible oxidative mechanism for the genotoxicity observed in the 1–3 μm SWCNT. Consequently, this study has demonstrated that SWCNT genotoxicity is dependent on its secondary structure under experimental conditions and oxidative stress alone cannot account for the observed damage.

Reproducible electrochemical etching of silver probes with a radius of curvature of 20 nm for tip-enhanced Raman applications

The performance of tip-enhanced Raman spectroscopy (TERS) largely depends on probe optimisation. An electrochemical etch using nitric acid and ethanol produces sharp silver probes with radius of curvature between 20 and 60 nm. Optimisation also identified controllable tapers; rough or smooth. Boundary element method simulations comparing the response to 532 nm excitation of silver and gold probes reveal no discernable field enhancement at the gold apex, but strong localised enhancement is observed at the silver apex. The motivation for employing this method of etching silver probes can also be seen in the signal enhancement observed in preliminary TERS data presented.

A surface extended X-ray absorption fine structure study of tellurium adsorbed onto Si(100)

Abstract The adsorption of tellurium on Si(100) has been studied using surface extended X-ray adsorption fine structure (SEXAFS) and X-ray standing wave spectroscopy (XSW). This particular system is of interest due to its potential applicability in the surfactant aided growth of CdHgTeCdTeSi(100) based infra-red detectors. The Te Si (100) structure was generated by depositing a thick layer (∼ 100 A) of CdTe onto a clean Si (2 × 1) double domain surface, and annealing the sample to 350°C. This resulted is a ∼ 1 ML Te terminated surface where the (2 × 1) reconstruction was lost in favour of a (1 × 1) symmetry. X-ray absorption of the Te L3 edge (E = 4341 eV), with a photon energy range of 4440–4700 eV, was probed using a total yield detection scheme. The SEXAFS results indicated that the Te atoms sat in 2-fold bridge sites directly above a fourth layer Si atom. The corresponding bond length was measured to be 2.52 ± 0.05 A . The XSW measurements of the (400) reflection gave a coherent position of 1.63 ± 0.03 A and a coherent fraction of 0.65. This is consistent with the breaking of the SiSi dimers and thus could be an example of the phenomena of adsorbate-induced dereconstruction of the surface. These results are compared with those of Bennet et al. who examined a similar system using soft X-ray photoemission (SXPS) and the STM study of Yoshikawa et al.

Quantum Dots for Multiplexed Detection and Characterisation of Prostate Cancer Cells Using a Scanning Near-Field Optical Microscope

In this study scanning near-field optical microscopy (SNOM) has been utilised in conjunction with quantum dot labelling to interrogate the biomolecular composition of cell membranes. The technique overcomes the limits of optical diffraction found in standard fluorescence microscopy and also yields vital topographic information. The technique has been applied to investigate cell-cell adhesion in human epithelial cells. This has been realised through immunofluorescence labelling of the cell-cell adhesion protein E-cadherin. Moreover, a dual labelling protocol has been optimised to facilitate a comparative study of the adhesion mechanisms and the effect of aberrant adhesion protein expression in both healthy and cancerous epithelial cells. This study reports clear differences in the morphology and phenotype of healthy and cancerous cells. In healthy prostate epithelial cells (PNT2), E-cadherin was predominantly located around the cell periphery and within filopodial extensions. The presence of E-cadherin appeared to be enhanced when cell-cell contact was established. In contrast, examination of metastatic prostate adenocarcinoma cells (PC-3) revealed no E-cadherin labelling around the periphery of the cells. This lack of functional E-cadherin in PC-3 cells coincided with a markedly different morphology and PC-3 cells were not found to form close cell-cell associations with their neighbours. We have demonstrated that with a fully optimised sample preparation methodology, multiplexed quantum dot labelling in conjunction with SNOM imaging can be successfully applied to interrogate biomolecular localisation within delicate cellular membranes.

Mechanisms of cell–cell adhesion identified by immunofluorescent labelling with quantum dots: A scanning near-field optical microscopy approach

Scanning near-field optical microscopy (SNOM) has been employed to simultaneously acquire high-resolution fluorescence images along with shear-force atomic force microscopy from cell membranes. Implementing such a technique overcomes the limits of optical diffraction found in standard fluorescence microscopy and also yields vital topographic information. The application of the technique to investigate cell-cell adhesion has revealed the interactions of filopodia and their functional relationship in establishing adherens junctions. This has been achieved via the selective tagging of the cell adhesion protein, E-cadherin, by immunofluorescence labelling. Two labelling routes were explored; Alexa Fluor 488 and semiconductor quantum dots. The quantum dots demonstrated significantly enhanced photostability and high quantum yield making them a versatile alternative to the conventional organic fluorophores often used in such a study. Analysis of individual cells revealed that E-cadherin is predominantly located along the cell periphery but is also found to extend throughout their filopodia. We have demonstrated that with a fully optimised sample preparation methodology, quantum dot labelling in conjunction with SNOM imaging can be successfully applied to interrogate biomolecular localisation within delicate cellular membranes.

The electronic passivation properties of Si nanoislands on GaAs(110) step defects induced by cleaving

Scanning tunneling microscopy and spectroscopy (STS) have been used to investigate submonolayer Si depositions on clean cleaved GaAs(110). These studies focus on the effect of cleaving-induced step defects on GaAs(110), the resultant Si overlayer formation, and the electrical properties of the substrate/overlayer system formed at 280 °C. STS measurements at a clean step edge on GaAs(110) show that the Fermi level shifts towards midgap for both n and p type, while measurements performed on Si nanoislands at step edges show that the Fermi level reverts back towards its “ideal” position. Results indicate the passivation properties of Si on GaAs(110) when deposited under these conditions. Implications for use in laser facets are discussed.

An investigation of the properties of intimate InInxGa1-xAs(100) interfaces formed at room and cryogenic temperatures

Abstract The electrical, chemical and structural properties of the interfaces formed at room and low temperatures, between In and atomically clean In 53 Ga 47 As/InP(100) have been studied. Current-voltage measurements indicate that diodes formed at 80 K exhibit significantly higher Schottky barriers (φ b = 0.45 eV) than diodes formed at 294 K (φ b = 0.30 eV). The reactions occurring during the formation of In In 53 Ga 47 As/InP(100) interfaces at room and low temperatures have been investigated using Soft X-Ray photoemission spectroscopy. Our results show that metallisation at room temperature results in a predominantly three dimensional mode of growth, accompanied by the out-diffusion of As. Low temperature (125 K) metallisation appears to reduce clustering and inhibit As out-diffusion. Examination of the resulting interfaces by transmission electron microscopy confirms the more uniform nature of the metal layers formed at low temperature. These observations, in conjunction with the barrier heights measured by the I-V technique, are discussed in the context of currently supported models of Schottky barrier formation.

Role of Raman spectroscopy and surface enhanced Raman spectroscopy in colorectal cancer.

Colorectal cancer (CRC) is the fourth most common cancer in the United Kingdom and is the second largest cause of cancer related death in the United Kingdom after lung cancer. Currently in the United Kingdom there is not a diagnostic test that has sufficient differentiation between patients with cancer and those without cancer so the current referral system relies on symptomatic presentation in a primary care setting. Raman spectroscopy and surface enhanced Raman spectroscopy (SERS) are forms of vibrational spectroscopy that offer a non-destructive method to gain molecular information about biological samples. The techniques offer a wide range of applications from in vivo or in vitro diagnostics using endoscopic probes, to the use of micro-spectrometers for analysis of biofluids. The techniques have the potential to detect molecular changes prior to any morphological changes occurring in the tissue and therefore could offer many possibilities to aid the detection of CRC. The purpose of this review is to look at the current state of diagnostic technology in the United Kingdom. The development of Raman spectroscopy and SERS in clinical applications relation for CRC will then be discussed. Finally, future areas of research of Raman/SERS as a clinical tool for the diagnosis of CRC are also discussed.

The electronic and structural properties of the silicon-gallium arsenide(110) interface

Abstract The passivation properties of the Si GaAs(110) interface have been studied using scanning tunnelling microscopy/spectroscopy (STM/STS) and soft X-ray photoemission spectroscopy (SXPS). Silicon has been deposited at room temperature and STM images show the sub-monolayer growth of silicon islands on the GaAs substrate. The electrical properties of these islands together with the clean surface have been investigated using scanning tunnelling spectroscopy (STS). The spectroscopy clearly illustrates the difference in electrical properties between atomically flat regions of GaAs as compared to those containing defects or steps, i.e. where surface band bending occurs. We have investigated the use of sub-monolayer Si coverages to modify the electronic structure of the surface. Height variations of 3–4Aacross Si islands and 2Aacross steps on the GaAs surface have also been observed using the STM. STS spectra, collected simultaneously with the STM image, showed the Si to have semiconducting properties differing from that of crystalline Si and the GaAs substrate. Comparisons between the STM and STS results together with SXPS have provided a correlation between the structural, electrical and chemical nature of the Si/GaAs(110) interface.