Signal analysis of total internal reflection fluorescent speckle microscopy (TIR-FSM) and wide-field epi-fluorescence FSM of the actin cytoskeleton and focal adhesions in living cells

Fluorescent speckle microscopy (FSM) uses low levels of fluorescent proteins to create fluorescent speckles on cytoskeletal polymers in high‐resolution fluorescence images of living cells. The dynamics of speckles over time encode subunit turnover and motion of the cytoskeletal polymers. We sought to improve on current FSM technology by first expanding it to study the dynamics of a non‐polymeric macromolecular assembly, using focal adhesions as a test case, and second, to exploit for FSM the high contrast afforded by total internal reflection fluorescence microscopy (TIR‐FM). Here, we first demonstrate that low levels of expression of a green fluorescent protein (GFP) conjugate of the focal adhesion protein, vinculin, results in clusters of fluorescent vinculin speckles on the ventral cell surface, which by immunofluorescence labelling of total vinculin correspond to sparse labelling of dense focal adhesion structures. This demonstrates that the FSM principle can be applied to study focal adhesions. We then use both GFP‐vinculin expression and microinjected fluorescently labelled purified actin to compare quantitatively the speckle signal in FSM images of focal adhesions and the actin cytoskeleton in living cells by TIR‐FM and wide‐field epifluorescence microscopy. We use quantitative FSM image analysis software to define two new parameters for analysing FSM signal features that we can extract automatically: speckle modulation and speckle detectability. Our analysis shows that TIR‐FSM affords major improvements in these parameters compared with wide‐field epifluorescence FSM. Finally, we find that use of a crippled eukaryotic expression promoter for driving low‐level GFP‐fusion protein expression is a useful tool for FSM imaging. When used in time‐lapse mode, TIR‐FSM of actin and GFP‐conjugated focal adhesion proteins will allow quantification of molecular dynamics within interesting macromolecular assemblies at the ventral surface of living cells.     

Efficient allylation of cellulose in dimethyl sulfoxide/tetrabutylammonium fluoride trihydrate

The cellulose solvent dimethyl sulfoxide (DMSO)/tetrabutylammonium fluoride trihydrate (TBAF) was successfully applied as reaction medium for the synthesis of allyl cellulose by conversion of the polymer with allyl chloride in the presence of solid NaOH. Samples with degree of substitution from 0.50 to 2.98 were accessible by varying the molar ratio anhydroglucose unit:allyl chloride:NaOH and reaction time. DMSO/TBAF was found to be an efficient reaction medium for the preparation of highly functionalized samples from spruce sulfite pulp with degree of polymerization of about 500 even in a scale of 50 g. The allyl cellulose samples were characterized by means of FTIR- and NMR spectroscopy. Size exclusion chromatography revealed negligible polymer degradation during synthesis and purification of the samples.     

Molecular weight effect on the obtainment of parallel and perpendicular orientation in thermotropic poly(diethylene glycol <Emphasis Type="Italic">p,p′</Emphasis>-bibenzoate)

Summary The effect of molecular weight on the uniaxial orientation process has been analyzed in two samples of thermotropic poly(diethylene glycol p,p′-bibenzoate), evaluating the influence of that parameter on the type of orientation obtained. Several strain rates and deformation temperatures have been tested in order to map out the conditions for obtaining the two different kinds of orientation. The results show that in the lower molecular weight PDEB sample (M_w=31200) it is rather easy to get exclusively perpendicular “anomalous” orientation, with the molecular axes aligned transversely to the stretching direction. However, it is extremely difficult to obtain 100% parallel “normal” orientation. On the contrary, either type of orientation or a mixture of them can be easily developed for the higher molecular weight PDEB sample (M_w=102900).     

X-ray topography studies of dislocations in single crystal CVD diamond

X-ray topography has been used to study single crystal diamond samples homoepitaxially grown by microwave plasma-assisted chemical vapour deposition (CVD) on high pressure high temperature (HPHT) and CVD synthetic diamond substrates. Clusters of dislocations in the CVD diamond layers emanated from points at or near the interface with the substrate. The Burgers vectors of observed dislocations have been determined from sets of {111} projection topographs. Dislocations have line directions close to the [001] growth direction and are either edge or 45° mixed dislocations. Where groups of dislocations originated at isolated points they tended to be of the edge variety. Where the substrate surface was deliberately damaged before growth, two sets of dislocations were observed to have propagated from each line of damage and there was a tendency for dislocations to be of the 45° mixed variety with a component of their Burgers vector parallel to the polishing direction. It is demonstrated that X-ray topography can be used to deduce the growth history of CVD synthetic diamond samples produced in multiple growth stages.     

Drying Kinetics of a Porous Spherical Particle and the Inversion Temperature

A model is presented for drying of a single porous particle with superheated steam and humid air. Experimental data for spherical porous ceramic particle reported in the literature were used for the validation of the model. An inversion temperature at which the evaporation rates within superheated steam and humid air are equal was predicted. The effect of thermophysical properties of the particle (permeability 10^-14 - 10^-17 m², diameter 3 × 10^-3 - 10 × 10^-3 m) and operating variables (gas mass flux 0.26 - 0.78 kg m^-2 s^-1, drying agent temperature 120-200°C) is tested. The inversion temperature is shown to be affected by the thermophysical properties of the porous particle and of the drying agent.     

Synthesis of TiO<Subscript>2</Subscript> Hybrid Molecular Imprinted Polymer for Ethofumesate Linked by Silane Coupling Agent

TiO₂ hybrid molecular imprinted polymer (MIP) for ethofumesate using methacrylic acid (MAA) as the functional monomer and silane coupling agent 3-(trimethoxysilyl) propylmethacrylate (KH570) as organic–inorganic connective bridge was synthesized via photo-excitation method. Hydrogen bond was proved to act between MAA and ethofumesate for pre- and post-polymerization binding properties as testified by UV spectrometric method. KH570 modified TiO₂ nanoparticles were prepared via sonochemical reaction, which can accelerate hydrolysis, increase collision chance for the reactive system and improve the dispersion of the nanoparticles. Scanning electron microscope (SEM), transmission electron microscope (TEM), binding and the adsorption kinetics experiments as well as thermogravimetric analysis (TGA) were employed for characterization. The results indicated that the hybrid MIP revealed a larger surface area and more ordered imprinting cavities with improved thermal stability compared to organic-only MIP. Furthermore, faster adsorption kinetics and enhancive adsorption capacity were achieved, which made it promising in chemical sensor applications.     

Luminescence from praseodymium doped AlN thin films deposited by RF magnetron sputtering and the effect of material structure and thermal annealing on the luminescence

Thin films of Praseodymium doped AlN are deposited on silicon (111) substrates at 77 K and 950 K by rf magnetron sputtering method. About 500–1000 nm thick films are grown at 100–200 watts RF power and 5–8 mTorr nitrogen, using a metal target of Al with Pr. X-rays diffraction results show that films deposited at 77 K are amorphous and those deposited at 950 K are crystalline. Cathodoluminescence studies are performed at room temperature and luminescence peaks are observed in a wide range from ultraviolet to infrared region. The most intense peak is obtained in green at 526 nm from amorphous films as a result from ³P₁→³H₅ transition. In crystalline films the intense peak was obtain in red at 648 nm as a result from ³P₀→³F₂ transition. Films are thermally activated at 1300 K for half an hour in a nitrogen atmosphere. Thermal activation enhances the intensity of luminescence. Two peaks at 488 nm and 505 nm merged after thermal activation, giving rise to a single peak at 495 nm.     

Effect of heat treatment in air on the thermal properties of SiC fibre reinforced composite: Part 2 A magnesium aluminium silicate (MAS) matrix glass ceramic composite

The thermal properties of a magnesium aluminium silicate (MAS) glass ceramic matrix composite reinforced by SiC (Nicalon) fibres have been investigated before and after heat treatment in the temperature range 600–1,200 °C. Within this temperature range, during the heat treatment at lower temperatures such as 600 and 700 °C, the oxidation of the carbon layer occurred and mixture of silicon and carbon was formed in the interface. This results in a decrease in thermal diffusivity values. After heat treatment at the temperatures higher than 1,000 °C, the carbon layer was thickened and resulted in the higher thermal diffusivity values.     

Optimizing simultaneously over the numerator and denominator polynomials in the Youla-Kuc/spl caron/era parametrization

Traditionally, when approaching controller design with the Youla-Kuc/spl caron/era parametrization of all stabilizing controllers, the denominator of the rational parameter is fixed to a given stable polynomial, and optimization is carried out over the numerator polynomial. In this note, we revisit this design technique, allowing to optimize simultaneously over the numerator and denominator polynomials. Stability of the denominator polynomial, as well as fixed-order controller design with H/sub /spl infin// performance are ensured via the notion of a central polynomial and linear matrix inequality (LMI) conditions for polynomial positivity.     

Measurement of the density and surface tension of Ni-based superalloys in the liquid and mushy states

The densities of three Ni-based superalloys have been measured in both liquid and mushy states by both a modified sessile drop method (MSDM) and a modified pycnometric method (MPM) for alloys CMSX-4 and CM186LC, and for CMSX-10 alloy by MSDM only. The surface tensions of liquid CMSX-4, CM186LC, and CMSX-10 superalloys were measured using the sessile drop method. All measurements were carried out in a highly purified argon atmosphere with the oxygen partial pressure of less than 10⁻¹⁹ MPa in the gas outlet. The densities of all superalloys in both liquid and mushy states were found to decrease with increasing temperature. The volume thermal expansion of each superalloy in the mushy state was found to be higher than that in the liquid state. The densities determined by different methods have been critically assessed and recommended values in both liquid and mushy states are given as a linear function of temperature for the three Ni-based superalloys. The surface tension of liquid CMSX-4 superalloy was found to decrease with increasing temperature, while that of liquid CMSX-10 superalloy increases with increasing temperature. The wetting behavior of liquid CM186LC on the alumina substrate was found (1) to differ significantly from that of CMSX-4 and CMSX-10 and (2) to vary with time. A HfO₂-rich layer was found in the contact area of CM186LC with the alumina substrate, which could lead to some uncertainty in the value obtained for the surface tension determined for CM186LC.