Application of mixed self-assembled monolayers (Mixed SAMs) for nucleic acid detection

Mixed self-assembled monolayers (Mixed SAMs) consisting of 8-Ferrocenyl-1-octanethiol and 6-Mercapto-1-hexanol (FcOT:MCH) with probe PNA on gold electrodes were fabricated by using two-step after the optimization of immobilization temperature of FcOT:MCH SAMs. Using AC voltammetry, a novel nucleic acid detection platform, with mixed SAMs, was proposed. A negative formal potential shift was observed after complementary ssDNA hybridization while there was no significant difference after non-complementary ssDNA hybridization. Compatible results were obtained with the measurement of formal potential differences between mixed SAMs and target DNA (complementary and non-complementary DNA) in different target DNA concentrations. The formal potential difference between mixed SAMs immobilization and complementary ssDNA hybridization was measured in different ionic strength concentrations.     

Direct atomic force microscopy observations of monovalent ion induced binding of DNA to mica

Multivalent ions in solution are known to mediate attraction between two like‐charged molecules. Such attraction has proved useful in atomic force microscopy (AFM) where DNA may be immobilized to a mica surface facilitating direct imaging in liquid. Theories of DNA immobilization suggest that either ‘salt bridging’ or fluctuation in the positions of counter ions about both the mica surface and DNA backbone secure DNA to the mica substrate. Whilst both theoretical and experimental evidence suggest that immobilization is possible in the presence of divalent ions, very few studies identify that such immobilization is possible with monovalent ions. Here we present direct AFM evidence of DNA immobilized to mica in the presence of only monovalent ions. Our data depict E. coli plasmid pBR322 adsorbed onto the negatively charged mica both after short (10 min) and long (24 h) incubation periods. These data suggest the need to re‐explore current theories of like‐charge attraction to include the possibility of monovalent interactions. We suggest that this DNA immobilization strategy may offer the potential to image natural processes with limited immobilization forces and hence enable maximum conformational freedom of the immobilized biomolecule.     

An analysis of heat transfer in the thin-layer transducer used for the measurements of EHD contact parameters

Thin-layer transducers, which are mainly used for the measurement of the EHD contact parameters, have to be attached to an isolating layer separating it from the steel roller. The isolating layer, made of SiO_x or Al₂O_3, disturbs heat flow in relation to the real contact. This paper presents a solution for the system of two heat conduction equations for a two-layer body: SiO_x-steel. The boundary conditions reflect the real situation during the measurements carried out by means of these transducers. The solution was obtained for an assumed temperature distribution of the isolating layer surface. Computation results for two different temperature distributions of the surface and for various values of the SiO_x-layer thickness are also presented.     

Reflectance in situ hybridization (RISH): detection,by confocal reflectance laser microscopy,of gold-labelled riboprobes in breast cancer cell lines and histological specimens

A method for reflectance in situ hybridization (RISH) is presented. The importance of the method is demonstrated by results obtained on cytological and histological breast cancer specimens. Scattering reflectance signals from 1-nm colloidal-gold particles after RNA/RNA in situ hybridization, using digoxigenin-labelled riboprobes, were detected by confocal scanning laser microscopy. The mRNA expression of two ras-related genes, rho B and rho C, was analysed in human histological breast cancer specimens and in human breast cancer cell lines. Horizontal (x, y) and vertical (z) optical sections after three-dimensional imaging were used for visualization. A marked heterogeneity (between individual cells and between specimens) was noted for the expression of the rho B gene, both in cytological and in histological samples. On the other hand, rho C was always expressed and showed no heterogeneity. This method allows the identification of several cellular constituents in an heterogeneous tissue structure, as demonstrated by the simultaneous detection of rho B (or rho C) by reflectance and of DNA, cytokeratin and/or vimentin by fluorescence.     

Immobilization of iron storage protein on a gold electrode based on self-assembled monolayers

Ferritin is a globular protein consisting of 24 subunits to form a hollow shell and is capable of storing iron in the cavity. Findings that the naturally existing iron core of ferritin can be readily extracted and replaced with a variety of electroactive materials make ferritin suitable for biosensor and biofuel cell applications. The immobilization of ferritin on the electrode surface is essential for various bioelectronic applications. In this work, based on self-assembled monolayers, ferritin was immobilized on a gold electrode through two different methods: chemisorption of thiolated ferritin onto bare gold electrodes and covalent binding of ferritin to succinimidyl alkanedisulfide-modified Au electrodes. Effects of experimental conditions on the ferritin immobilization were investigated. The ferritin immobilized on the gold electrode was characterized by atomic force microscopy and cyclic voltammetry.     

GaAs纳米线阵列光阴极的制备及其特性研究

采用改进的Stber法合成直径为500nm的SiO_2纳米球,用旋涂法将所合成的SiO_2纳米球制作掩模版,采用纳米球刻蚀法制备GaAs纳米线阵列。利用扫描电子显微镜(SEM)、漫反射谱对其进行了表征分析,再对所制备出的GaAs纳米线阵列结构进行Cs-F交替激活实验,使其表面形成负电子亲和势光阴极,并对最终制备出的GaAs纳米线阵列光阴极样品进行量子效率测试,验证了GaAs纳米线阵列结构的量子效率比GaAs基片提高50%以上,从而证实了纳米线阵列结构的高光电转换效率。     

High‐contrast optical microscopy of graphene sheets

In the way of making graphene an industry‐friendly material, it must be mass‐produced with high‐quality and reduced cost over large areas. Assisted by machine‐learning techniques, rapid, nondestructive and accurate determination of large graphene sheets on SiO₂/Si substrates has been made possible in recent years by the optical microscopy method. Optimization of the substrate to achieve the maximum contrast can further extend the application of the optical microscopy method for quality control of the mass‐produced graphene. Graphene/n₂/n₃three‐layer structures, where n₂ and n₃ are refractive indices, are routinely used for identifying the number of graphene layers by optical reflection microscopy. In this paper, two analytical equations are derived that can be easily used for high‐contrast optical imaging of graphene sheets without any need to resort to the cumbersome numerical methods. One of the equations is derived for choosing the best material with refractive index n₂ that when coated on a substrate with refractive index n₃, maximizes the optical contrast. The other equation is derived for finding the best thickness of the SiO₂ layer in graphene/SiO₂/Si structures, which are in common use for fabrication of graphene‐based devices. The results are implemented in a MATLAB GUI, see Supporting Information, to assist the users in using the equations.     

EFFECT OF SURFACE ENERGY ON THE GROWTH OF DIAMOND-LIKE CARBON/AMORPHOUS SILICON FILMS ON VARIOUS SUBSTRATES

Diamond-like carbon/amorphous silicon bilayer films were deposited on SiO₂, Ge, and Ta₂O₅ substrates using a pulsed filtered cathodic arc (PFCA) system. Amorphous silicon (a-Si) layer was firstly deposited on three substrates using DC magnetron sputtering, then diamond-like carbon (DLC) film was deposited on a-Si layer via pulsed filtered cathodic arc. The thicknesses of a-Si layer and DLC film as monitored by in-situ ellipsometry during the film deposition were 7 and 10 nm, respectively. The surface energy of SiO₂, Ge, and Ta₂O₅ substrates was determined by measuring the contact angle of water on these substrates. It was found that the contact angles of water on SiO₂, Ge, and Ta₂O₅ substrates were 53°, 63°, and 75°, respectively. This result indicates that SiO₂ has the highest surface energy while Ta₂O₅ has the lowest surface energy. The thickness of the a-Si layer and DLC film was determined from the cross-section transmission electron microscopy (TEM) images. The thinnest a-Si layer of 5.64 nm was obtained from SiO₂ substrate which has the highest surface energy. The thickest a-Si layer of 6.97 nm was obtained from Ta₂O₅ corresponding to the lowest surface energy. This study shows that the thickness of the growth film strongly depends on the surface energy of the substrate. However, the DLC films deposited on each a-Si layer of three substrates have the same thickness approximately of 9.9 nm, because all of them were deposited on a-Si layers having the same surface energy.     

Effect of Interfacial Properties on EHL Under Pure Sliding Conditions

This paper presents an experimental study of the effect of boundary slip on the lubricating film shape and friction of an elastohydrodynamic lubrication (EHL) contact under isothermal conditions. Ball and disc pure sliding experiments were carried out with a high viscosity polybutene oil using a conventional optical EHL test rig. The film shape and friction were measured simultaneously. The results obtained from two discs with different coatings were compared. One disc was coated only with Cr, the partially reflective layer, and the other had an extra layer of SiO₂ coating on top. When running under mild conditions of low load and speed, there was no evidence of any boundary slip effect. However, when the load increased, the Cr-coated disc produced lower film thickness and friction than the SiO₂-coated disc. The Cr-coated surface had a larger contact angle, i.e., smaller surface energy, than the SiO₂ surface, which reflects the weak bonding between the molecules of the surface and the lubricant. The study concludes that surfaces with low surface energy promote boundary slip at the EHL contact, leading to a reduction in friction and film thickness.     

Characterization of lacquer films from the middle and late Chinese warring states period 476–221BC

The famous lacquer wares excavated from the Jiuliandun Tombs of the middle and late Warring States period (476–221 BC) were analyzed using scanning electron microscopy/energy as well as dispersive X‐ray spectrometry (SEM/EDS), Raman spectroscopy (RS), optical microscopy (OM), and X‐ray diffraction (XRD). The results showed a multilayer structure in the lacquer film, including a Qihui layer (a layer made of lacquer and various plasters), undercoat layer (or finishing coat) and colored paint layer mixed with various inorganic particles, such as quartz (SiO₂) and hydroxyapatite [Ca₅(PO₄)₃(OH)], as fillers in the Qihui layers or orpiment (As₂S₃) and cinnabar (HgS), which were used as a yellow or red pigment, respectively. With the help of elemental mapping images, a double‐layer structure of the lacquer plaster was observed, corresponding to a mixture of lacquer liquid and bone ash [Ca₅(PO₄)₃(OH)], with large‐diameter particles in the ground lacquer layer near the wooden body and small quartz (SiO₂) particles in upper lacquer layer. Specifically, quartz particles detected in the undercoat layer as fillers could be beneficial for improving the moshardness value, cost reduction and abrasive resistance of the lacquer film. In fact, the mixed method that used urushi and inorganic particles to form lacquer plaster was an important technological innovation and deeply influenced lacquering technologies worldwide. The results of this study will not only contribute to understanding the importance of lacquer skills in the Chinese Warring States but also provide information for cultural relic conservation as well as modern lacquer manufacturing for their protection and duplication.     

Characterisation,by the PIXE Method,of Trace Elements During Physicochemical Reactions at the Periphery of Bioactive Glass Pastilles in Contact with Biological Fluids

Abstract

The prerequisite for bioactive glasses to bond to living bone is the formation of biologically active apatites on their surface in the body. Reactions and bioactivity mechanisms between bioactive glasses and bone depend on the glass composition. We study a glass in the SiO₂‐Na₂O‐CaO‐P₂O₅‐K₂O‐Al₂O₃‐MgO system. To characterise physicochemical reactions at the materials periphery, we immersed the glass pastilles into biological fluids for periods of 5, 10, and 20 days. The surface changes were studied at the micrometer scale by a Particle Induced X‐ray Emission (PIXE) method associated with Rutherford Backscattering Spectroscopy (RBS). After 20 days of immersion, elemental maps showed the formation of a calcium‐phosphate layer at the surface of the glass pastilles. The thickness of this layer was around 15 µm. Thanks to the PIXE method, we demonstrated the presence of traces of Mg in this layer. The glass doped with MgO leads to the formation of an apatite which incorporates magnesium. Formation of this Ca‐P‐Mg layer represents the bioactive properties of the studied glass. This biologically active layer improves the properties of the glass and will permit a chemical bond between the ceramic and bone.     

新型法布里-珀罗（Fabry—Perot）腔传感结构

应用化学腐蚀和熔接技术对光纤端面进行腐蚀与熔接的方法,构建法布里-珀罗（F-P）腔,根据法布里-珀罗腔的干涉原理设计光纤传感结构,并应用紫外交联法在传感头固定化生物物质RNA,通过观测固定化前后端面SEM图像的对比与F-P腔干涉图谱的相位偏移,得出光纤端面RNA的固定化隋况。     

AFM imaging of functionalized double-walled carbon nanotubes

We present a comparative study of several non-covalent approaches to disperse, debundle and non-covalently functionalize double-walled carbon nanotubes (DWNTs). We investigated the ability of bovine serum albumin (BSA), phospholipids grafted onto amine-terminated polyethylene glycol (PL-PEG₂₀₀₀-NH₂), as well as a combination thereof, to coat purified DWNTs. Topographical imaging with the atomic force microscope (AFM) was used to assess the coating of individual DWNTs and the degree of debundling and dispersion. Topographical images showed that functionalized DWNTs are better separated and less aggregated than pristine DWNTs and that the different coating methods differ in their abilities to successfully debundle and disperse DWNTs. Height profiles indicated an increase in the diameter of DWNTs depending on the functionalization method and revealed adsorption of single molecules onto the nanotubes. Biofunctionalization of the DWNT surface was achieved by coating DWNTs with biotinylated BSA, providing for biospecific binding of streptavidin in a simple incubation step. Finally, biotin-BSA-functionalized DWNTs were immobilized on an avidin layer via the specific avidin–biotin interaction.     

Interfacial reactions and silicate formation in highly dispersed Lu2O3– SiO2 system

Solid state interface reactions in highly dispersed Lu₂O₃– SiO₂ binary oxide system were studied at 600–1100 °C with X‐ray powder diffraction (XRD), high‐resolution transmission electron microscopy (HRTEM) and Fourier Transform Infrared spectroscopy (FTIR). The results show that at 600–900 °C an amorphous, nanometer thick Lu‐O‐Si layer covering SiO₂ particles exists in the system. At higher temperatures the breakage of the layer into amorphous islands occurs and crystalline silicates with various structures are formed. In particular, Lu₄[Si₃O₁₀][SiO₄] silicate, analogue of B‐type Dy – Tm disilicates, forms at 1000 °C.     

Development and application of copper–nickel zirconium diboride as EDM electrodes manufactured by selective laser sintering

The production of electrical discharge machining (EDM) electrodes by conventional machining processes can account for over 50 % of the total EDM process costs. The emerging additive manufacturing (AM) technologies provide the possibility of direct fabrication of EDM electrodes. Selective laser sintering (SLS) is an alternative AM technique because it has the possibility to reduce the tool-room lead time and total EDM costs. The main difficulty of manufacturing an EDM electrode using SLS is the selection of an appropriate material. This work investigated the direct production of EDM electrodes by means of the SLS using a newly developed non-conventional metal–matrix composite material composed of a metallic matrix (CuNi) and an advanced ceramic (ZrB₂). The influence of important SLS parameters and material content on the densification behavior and porosity of the electrodes was investigated. EDM experiments were conducted to observe the electrodes behavior and performance. It was found that the ZrB₂-CuNi electrodes could be successfully manufactured by SLS. Interlayer bonding and porosity are directly influenced by the layer thickness. Smaller layer thicknesses improved bonding between layers and decreased the porosity of the parts. The laser scan speed has a significant effect on the densification behavior. The scan line spacing affects the pore structure by means of overlapping. The surface morphology of the samples was not affected by varying the scan line spacing. The ZrB₂-CuNi electrodes presented a much superior performance than SLS copper powder electrodes, but inferior to solid copper electrodes.     

Molecular dynamics simulation on the tribology properties of two hard nanoparticles (diamond and silicon dioxide) confined by two iron blocks

The tribology behaviors of diamond and silicon dioxide (SiO₂) nanoparticles were examined via molecular dynamics simulations; four cases were simulated. At low velocity and low load, the nanoparticles separated the two blocks from each other and acted as ball-bearings. The plastic deformation, temperature distribution, and friction force were all improved due to the action of the nanoparticles. However, the crushing of the SiO₂ nanoparticles was accompanied by deformation-induced loss of the rolling effect, when the load was increased. Without nanoparticles, a transfer layer formed at high velocity and low load. The two nanoparticles provided support for a certain duration. However, at high velocity and high load, the support effect of these nanoparticles was lost in a short sliding time.     

Detection of single copy sequences using BAC-FISH and C-PRINS techniques in sunflower chromosomes

Bacterial artificial chromosome - fluorescence in situ hybridization (BAC-FISH) and cyclingprimed in situ labeling (C-PRINS) techniques were evaluated for integration of physical and genetic maps of sunflower (Helianthus annuus L.). Single-site SSR markers were selected from three linkage groups of a high-density sunflower genetic map. This selection was based on previously identified QTL associated to S. sclerotiorum. These markers were used to select BACs contaning single copy sequences for BAC-FISH aplication. Blocking of highly dispersed repetitive sunflower sequences reduced unspecific hybridization, and allowed the detection of specific signals for BACs containing SSR markers HA4222 and HA2600, anchored to LG 16 and LG 10, respectively. Single-site FISH signal detection was optimized by adjusting the relative quantity and quality of unlabelled repetitive sequences present in the blocking DNA. The SSR marker ORS1247 anchored to the LG 17 was detected by C-PRINS, which yielded fluorescence signals that were specific and intense. This progress in localizing single-copy sequences using BAC-FISH and indirect CPRINS strategies in sunflower will facilitate the integration of genetic and physical maps, allowing the identification of     

Ion microprobe studies of reactions in squeeze-cast aluminium alloy matrix composites

An ion microprobe with high lateral resolution has been used to study the chemical reactions at the fibre/matrix interface of metal–matrix composites. During the squeeze-casting process, the Al–Si–Mg matrix reacts with the preform made of Saffil fibres (96% Al₂O₃, 4% SiO₂). The reaction occurs mainly between the silica binder and Mg from the matrix according to SiO₂ + 2Mg = 2MgO + Si. A continuous layer of MgO was formed around the fibres, even on surfaces that were not covered by the silica binder. Possible reasons are discussed for the formation of MgO in areas where binder coating was missing. In such areas, Mg reduces SiO₂ that is contained in the fibre. However, the fibres (A1₂O₃) are not attacked by Mg. In the isolated case of fibres that were completely uncoated, no reaction products were observed at the interface. The presence of silica binder seems to be an essential requirement for this reaction to occur. When squeeze-casting is performed with sufficiently high melt temperature, Al from the matrix also reduces silica.     

Wear-resistance mechanism of an Al–12Si alloy reinforced with aluminosilicate short fibers

The wear behavior of an aluminosilicate (Al₂O₃·SiO₂) short-fiber-reinforced Al–12Si alloy composite and the parent Al–12Si alloy were investigated under dry conditions. The results show that the increased wear resistance of Al₂O₃·SiO₂/Al–12Si can be attributed to the formation of a hardened layer in the sub-surface region where realignment and redistribution of fragmented eutectic phase and fragmented aluminosilicate fibers occur during dry sliding.     

Atomic force microscopy tips (cantilevers) as molecular nucleic acid sensors

A model single strand DNA (ssDNA) was covalently immobilized onto AFM tips (cantilevers) as specific ligand. These tips were interacted with the buffer solutions with or without free ssDNA molecules as the target strands to be detected. Immobilization and hybridization onto the cantilever surfaces were observed by optical and fluorescence microscopies. Interactions between the AFM tip (cantilever) and the aqueous medium (therefore with the target ssDNAs) were quantified by obtaining the "percent separation distance" ("PSD") as the main variable. The PSD values obtained for the buffer solutions were between -2.07 and +4.91%. There were slight increases in the negative values when non-complementary ssDNA molecules were introduced into the buffer. However, after hybridization with its complementary ssDNA, the PSD values were significantly increased (between -32.24 and -43.47%). There was a correlation between the concentration of the complementary target ssDNA in the medium and the PSD value. As a result of these promising results it was concluded that this approach may be further developed to create AFM-based molecular sensors for diverse applications.