Multiplexed tandem polymerase chain reaction identifies strong expression of oestrogen receptor and Her-2 from single, formalin-fixed, paraffin-embedded breast cancer sections

Aim To establish the suitability of multiplex tandem polymerase chain reaction (MT-PCR) for rapid identification of oestrogen receptor (ER) and Her-2 status using a single, formalin-fixed, paraffin-embedded (FFPE) breast tumour section. Methods Tissue sections from 29 breast tumours were analysed by immunohistochemistry (IHC) and fluorescence in situ hybridisation (FISH). RNA extracted from 10μm FFPE breast tumour sections from 24 of 29 tumours (14 ER positive and 5 Her-2 positive) was analysed by MT-PCR. After establishing a correlation between IHC and/or FISH and MT-PCR results, the ER/Her-2 status of a further 32 randomly selected, archival breast tumour specimens was established by MT-PCR in a blinded fashion, and compared to IHC/FISH results. Results MT-PCR levels of ER and Her-2 showed good concordance with IHC and FISH results. Furthermore, among the ER positive tumours, MT-PCR provided a quantitative score with a high dynamic range. Threshold values obtained from this data set applied to 32 archival tumour specimens showed that tumours strongly positive for ER and/or Her-2 expression were easily identified by MT-PCR. Conclusion MT-PCR can provide rapid, sensitive and cost-effective analysis of FFPE material and may prove useful as triage to identify patients suited to endocrine or trastuzumab (Herceptin) treatment.

Localization of fibronectin in the rat testis : effects of serum and dibutyryl cyclic AMP on fibronectin deposition by peritubular myoid cells in culture

The peritubular zone of the rat testis has an extensive extracellular matrix (ECM). Fibronectin (FN) is distributed primarily in the basal lamina of the seminiferous tubule boundary tissue and is synthesized by peritubular myoid cells. Several extracellular changes are mediated by growth factors and these changes occur at the time of hormone mediated testicular development, particularly in the peritubular zone. The effects of serum or dibutyryl cyclic AMP (cAMP) on FN production by the mesenchymal peritubular myoid cells were evaluated. Rats of various ages (10, 15, 20, 40 and 80 days) were employed for immunofluorescent localization of rat testicular FN in frozen sections. In all age groups tested, FN was primarily present in a broad layer around each seminiferous tubule, and blood vessel, and in variable distribution throughout the interstitial stroma. By day 20 there was no clear distinction in FN staining between the peritubular zone and the interstitial tissue. This indicates an involvement of FN in the ECM developments which occur in the peritubular zone of the testis at this time. The peritubular myoid cells were isolated from 20-22 day old rat testis and cultured on glass coverslips. These cells were grown to confluence with 10% fetal calf serum (FCS) in medium until day 4 and then subcultured to have secondary monocultures maintained with or without serum. By means of immunofluorescence and cytochemistry using avidin-biotin peroxidase complex it was observed that peritubular myoid cells were positive for FN and most of the FN was localized in the perinuclear region. Subcultured peritubular myoid cells maintained for 4 days in medium containing FCS developed an extensive interconnecting FN matrix. In the presence of 0.5 mM cAMP in culture, FN became localized along the filamentous process of peritubular myoid cells and more prominently in the areas of triangulated multi-cell aggregates as well as on the surface of the contracted small spherical cells. The addition of cAMP in the presence of FCS, also caused a noticeable change in the staining pattern; FN was detected along the filamentous process developing into a complex network of cells encased in an extensive matrix. It would appear that the translocation of FN in the cytoplasmic extensions of peritubular myoid cells may be a direct consequence of morphological changes associated with metabolic regulation of cAMP. This may also be related to the puberty associated development of in vivo changes in the ECM produced by peritubular myoid cells.

Characteristics of ultrafine particle sources and deposition rates in primary school classrooms

The aim of this work was to investigate changes in particle number concentration (PNC) within naturally ventilated primary school classrooms arising from local sources either within or adjacent to the classrooms. We quantify the rate at which ultrafine particles were emitted either from printing, grilling, heating or cleaning activities and the rate at which the particles were removed by both deposition and air exchange processes. At each of 25 schools in Brisbane, Australia, two weeks of measurements of PNC and CO2 were taken both outdoors and in the two classrooms. Bayesian regression modelling was employed in order to estimate the relevant rates and analyse the relationship between air exchange rate (AER), particle infiltration and the deposition rates of particle generated from indoor activities in the classrooms. During schooling hours, grilling events at the school tuckshop as well as heating and printing in the classrooms led to indoor PNCs being elevated by a factor of more than four, with emission rates of (2.51 ± 0.25) x 1011 p min-1, (8.99 ± 6.70) x 1011 p min-1 and (5.17 ± 2.00) x 1011 p min-1, respectively. During non-school hours, cleaning events elevated indoor PNC by a factor of above five, with an average emission rate of (2.09 ± 6.30) x 1011 p min-1. Particles were removed by both air exchange and deposition; chiefly by ventilation when AER > 0.7 h-1 and by deposition when AER < 0.7 h-1.

Formation and electron field emission of graphene films grown by hot filament chemical vapor deposition

Graphene films with different structures were catalytically grown on the silicon substrate pre-deposited with a gold film by hot filament chemical vapor deposition under different conditions, where methane, hydrogen and nitrogen were used as the reactive gases. The morphological and compositional properties of graphene films were studied using advanced instruments including field emission scanning electron microscopy, micro-Raman spectroscopy and X-ray photoelectron spectroscopy. The results indicate that the structure and composition of graphene films are changed with the variation of the growth conditions. According to the theory related to thermodynamics, the formation of graphene films was theoretically analyzed and the results indicate that the formation of graphene films is related to the fast incorporation and precipitation of carbon. The electron field emission (EFE) properties of graphene films were studied in a high vacuum system of ∼10-6 Pa and the EFE results show that the turn-on field is in a range of 5.2-5.64 V μm-1 and the maximum current density is about 63 μ A cm-2 at the field of 7.7 V μm-1. These results are important to control the structure of graphene films and have the potential applications of graphene in various nanodevices.

Room-temperature photoluminescence from nitrogenated carbon nanotips grown by plasma-enhanced hot filament chemical vapor deposition

Nitrogenated carbon nanotips with a low atomic concentration of nitrogen have been synthesized by using a custom-designed plasma-enhanced hot-filament plasma chemical vapor deposition system. The properties (including morphology, structure, composition, photoluminescence, etc.) of the synthesized nitrogenated carbon nanotips are investigated using advanced characterization tools. The room-temperature photoluminescence measurements show that the nitrogenated carbon nanotips can generate two distinct broad emissions located at ∼405 and ∼507 nm, respectively. Through the detailed analysis, it is shown that these two emission bands are attributed to the transition between the lone pair valence and bands, which are related to the sp3 and sp2 C-N bonds, respectively. These results are highly relevant to advanced applications of nitrogenated carbon nanotips in light emitting optoelectronic devices.

Low-pressure planar magnetron discharge for surface deposition and nanofabrication

Current-voltage characteristics of the planar magnetron are studied experimentally and by numerical simulation. Based on the measured current-voltage characteristics, a model of the planar magnetron discharge is developed with the background gas pressure and magnetic field used as parameters. The discharge pressure was varied in a range of 0.7-1.7 Pa, the magnetic field of the magnetron was of 0.033-0.12 T near the cathode surface, the discharge current was from 1 to 25 A, and the magnetic field lines were tangential to the substrate surface in the region of the magnetron discharge ignition. The discharge model describes the motion of energetic secondary electrons that gain energy by passing the cathode sheath across the magnetic field, and the power required to sustain the plasma generation in the bulk. The plasma electrons, in turn, are accelerated in the electric field and ionize effectively the background gas species. The model is based on the assumption about the prevailing Bohm mechanism of electron conductivity across the magnetic field. A criterion of the self-sustained discharge ignition is used to establish the dependence of the discharge voltage on the discharge current. The dependence of the background gas density on the current is also observed from the experiment. The model is consistent with the experimental results. © 2010 American Institute of Physics.

Inductively coupled plasma-assisted RF magnetron sputtering deposition of boron-doped microcrystalline Si films

An innovative custom-designed inductively coupled plasma-assisted RF magnetron sputtering deposition system has been developed to synthesize B-doped microcrystalline silicon thin films using a pure boron sputtering target in a reactive silane and argon gas mixture. Films were deposited using different boron target powers ranging from 0 to 350 W at a substrate temperature of 250 °C. The effect of the boron target power on the structural and electrical properties of the synthesized films was extensively investigated using X-ray diffraction, Raman spectroscopy, scanning electron microscopy, and Hall-effect system. It is shown that, with an initial increase of the boron target power from 0 to 300 W, the structural and electrical properties of the B-doped microcrystalline films are improved. However, when the target power is increased too much (e.g. to 350 W), these properties become slightly worse. The variation of the structural and electrical properties of the synthesized B-doped microcrystalline thin films is related to the incorporation of boron atoms during the crystallization and doping of silicon in the inductively coupled plasma-based process. This work is particularly relevant to the microcrystalline silicon-based p-i-n junction solar cells.

Inductively coupled plasma-assisted RF magnetron sputtering deposition of highly uniform SiC nanoislanded films

A new deposition technique-inductively coupled plasma-assisted RF magnetron sputtering has been developed to fabricate SiC nanoislanded films. In this system, the plasma production and magnetron sputtering can be controlled independently during the discharge. The deposited SiC nanoislanded films are highly uniform, have excellent stoichiometry, have a typical size of 10-45 nm, and contain small (∼ 6 nm) cubic SiC nanocrystallites embedded in an amorphous SiC matrix.

RF magnetron sputtering deposition of bioactive Ca-P-based coatings on Ti-6Al-4V alloy

RF magnetron concurrent sputtering of Hydroxyapatite and Ti forming functionally graded calcium phosphate-based composite bioactive films on Ti-6Al-4V orthopedic alloy is reported. Calcium oxide phosphate (4CaO•P2O5) is the main crystalline phase. In vitro cell culturing tests suggest outstanding biocompatibility of the Ca-P-Ti films. Images of the plasma-enhanced sputtering processes and cell culturing are presented and discussed.

Investigation on bioactivity of titanium dioxide thin films fabricated by electron cyclotron resonance plasma aided pulsed laser deposition

Titanium dioxide thin films with a rutile crystallinite size around 20 nm were fabricated by pulsed laser deposition (PLD) aided with an electron cyclotron resonance (ECR) plasma. With annealing treatment, the crystal size of the rutile crystallinite increased to 100 nm. The apatite-forming ability of the films as deposited and after annealing was investigated in a kind of simulated body fluid with ion concentrations nearly equal to those of human blood plasma. The results indicate that ECR aided PLD is an effective way both to fabricate bioactive titanium dioxide thin films and to optimize the bioactivity of titanium dioxide, with both crystal size and defects of the film taken into account.

Tailoring carbon nanotips in the plasma-assisted chemical vapor deposition: Effect of the process parameters

Carbon nanotips have been synthesized from a thin carbon film deposited on silicon by bias-enhanced hot filament chemical vapor deposition under different process parameters. The results of scanning electron microscopy indicate that high-quality carbon nanotips can only be obtained under conditions when the ion flux is effectively drawn from the plasma sustained in a CH4 + NH3 + H2 gas mixture. It is shown that the morphology of the carbon nanotips can be controlled by varying the process parameters such as the applied bias, gas pressure, and the NH3 / H2 mass flow ratios. The nanotip formation process is examined through a model that accounts for surface diffusion, in addition to sputtering and deposition processes included in the existing models. This model makes it possible to explain the major difference in the morphologies of the carbon nanotips formed without and with the aid of the plasma as well as to interpret the changes of their aspect ratio caused by the variation in the ion/gas fluxes. Viable ways to optimize the plasma-based process parameters to synthesize high-quality carbon nanotips are suggested. The results are relevant to the development of advanced plasma-/ion-assisted methods of nanoscale synthesis and processing.

Analysis of photoluminescence background of Raman spectra of carbon nanotips grown by plasma-enhanced chemical vapor deposition

Carbon nanotips with different structures were synthesized by plasma-enhanced hot filament chemical vapor deposition and plasma-enhanced chemical vapor deposition using different deposition conditions, and they were investigated by scanning electron microscopy and Raman spectroscopy. The results indicate that the photoluminescence background of the Raman spectra is different for different carbon nanotips. Additionally, the Raman spectra of the carbon nanotips synthesized using nitrogen-containing gas precursors show a peak located at about 2120 cm-1 besides the common D and G peaks. The observed difference in the photoluminescence background is related to the growth mechanisms, structural properties, and surface morphology of a-C:H and a-C:H:N nanotips, in particular, the sizes of the emissive tips.

Graphitization of nanocrystalline carbon microcoils synthesized by catalytic chemical vapor deposition

Graphitization, a common process involving the transformation of metastable nongraphitic carbon into graphite is one of the major present-day challenges for micro- and nanocarbons due to their unique structural character and highly unusual thermal activation. Here we report on the successful graphitization of nanocrystalline carbon microcoils prepared by catalytic chemical vapor deposition and post-treated in argon atmosphere at temperatures ∼2500 °C for 2 h. The morphology, microstructure, and thermal properties of the carbon microcoils are examined in detail. The graphitization mechanism is discussed by invoking a model of structural transformation of the carbon microcoils. The results reveal that after graphitization the carbon microcoils are prominently purified and feature a clear helical morphology, as well as a more regular and ordered microstructure. The interlayer spacing of the carbon microcoils decreases from 0.36 to 0.34 nm, whereas the mean crystal sizes in the c - and a -directions increase from 1.64 to 2.04 nm and from 3.86 to 7.21 nm, respectively. Thermal treatment also substantially improves the antioxidation properties of the microcoils by lifting the oxidation onset temperature from 550 to 672 °C. This process may be suitable for other nongraphitic micro- and nanomaterials.

Carbon nanofiber growth in plasma-enhanced chemical vapor deposition

A theoretical model to describe the plasma-assisted growth of carbon nanofibers (CNFs) is proposed. Using the model, the plasma-related effects on the nanofiber growth parameters, such as the growth rate due to surface and bulk diffusion, the effective carbon flux to the catalyst surface, the characteristic residence time and diffusion length of carbon atoms on the catalyst surface, and the surface coverages, have been studied. The dependence of these parameters on the catalyst surface temperature and ion and etching gas fluxes to the catalyst surface is quantified. The optimum conditions under which a low-temperature plasma environment can benefit the CNF growth are formulated. These results are in good agreement with the available experimental data on CNF growth and can be used for optimizing synthesis of related nanoassemblies in low-temperature plasma-assisted nanofabrication. © 2008 American Institute of Physics.

Inductively coupled Ar/CH₄/H₂plasmas for low-temperature deposition of ordered carbon nanostructures

The study of inductively coupled Ar/CH 4/H 2 plasmas in the plasma enhanced chemical vapor deposition (PECVD) of self-assembled carbon nanostructures (CN) was presented. A spatially averaged (global) discharge model was developed to study the densities and fluxes of the radical neutrals and charged species, the effective electron temperature, and methane conversion factors under various conditions. It was found that the deposited cation fluxes in the PECVD of CNs generally exceed those of the radical neutrals. The agreement with the optical emission spectroscopy (OES) and quadrupole mass spectrometry (QMS) was also derived through numerical results.