A simple handling technique for mammalian oocytes and embryos during preparation for transmission electron microscopy

Due to their small size, mammalian oocytes and embryos pose unique problems during preparation for transmission electron microscopy. This paper outlines a method which combines protein embedding with centrifugation to locate the specimens on the face of a Beem capsule mould. This method facilitates both the processing of oocytes with minimal loss and rapid location of the specimens within the block for simultaneous sectioning, staining and examination.     

Effects of gallia additions on sintering behavior of gadolinia-doped ceria

The effects of gallia additions on the sintering behavior of gadolinia-doped ceria were systematically investigated from the following aspects: the variation in sintered density, the variation in grain size, and the existing forms of Ga₂O₃ in CeO₂.Sintered density increased with increasing Ga₂O₃ content up to 5 mol.% and then it decreased with further addition of Ga₂O₃. Grain size also increased with increasing Ga₂O₃ content up to 5 mol.% and then decreased with further addition of Ga₂O₃. Decrease in grain size was caused by a pinning effect of Ga₂O₃ precipitation at grain boundaries. Lattice constant decreased with increasing Ga₂O₃ content up to 5 mol.%. This decrease will be due to the substitution of smaller Ga³⁺ ions for Ce⁴⁺ ions in the CeO₂ structure. According to the results obtained from scanning electron microscope (SEM) and X-ray diffraction (XRD) analyses, the solubility limit of Ga₂O₃ in Ce_0.8Gd_0.2O_1.9 ceramics can be estimated to be nearly 5 mol.%. The addition of Ga₂O₃ up to the solubility limit was found to promote the sintering properties of Gd₂O₃-doped CeO₂.     

Discrimination of B–C–N nanotubes through energy-filtering electron microscopy

Various multi-walled nanotubes in the B–C–N system are thoroughly investigated using a JEOL-3100FEF high-resolution field emission transmission electron microscope operating at 300 kV and equipped with an in-column built Omega filter. Spatially-resolved B, C and N elemental maps of the nanotubes are constructed. It is realized that a wide variety of tubular arrays composed of B, C and N atoms may exist in the system. Sandwich-like BN-rich and C-rich alternating tubular shells, graphitic C layers inside and outside of pure BN shells induced either by surface contamination, or electron beam irradiation, separation of C-rich and BN-rich tubes and/or BN particles within tubular bunches may take place. One should carefully take these effects into account while analyzing nanotube physical properties, e.g., electrical or optical, rather than simply rely on electron energy loss spectra typically collected from B, C and N containing nanostructures as a whole. Striking dependence of an individual nanotube electrical conductivity on tubular shell chemistry is demonstrated using I–V curve recording in an atomic force microscope.     

Assessing organo-clay dispersion in polymer layered silicate nanocomposites: A SAXS approach

A SAXS method for the quantitative assessment of the morphology of polymer layered silicate nanocomposites is proposed. Fitting the SAXS patterns, the number of clay layers, the periodicity of the layers in the tactoids, the thickness of the regions interposed between the clay platelets and their distributions can be measured. A good agreement with TEM data was obtained, avoiding the inconsistencies with microscopical observations often reported in the literature.     

Corrosion Behavior of 316LN and 316 Stainless Steels During Long-term Exposure to Aerated 0.5 M NaCl Using Electrochemical Noise Technique

In the present work 316LN as well as 316 stainless steel (SS) coupons each of dimensions (0.025 × 0.018 × 0.006 m³) in well-polished condition were used as two nominal electrodes which were immersed in the aerated solution of 0.5 M NaCl. Correlated current and potential electrochemical noise (EN) signals were collected at 1 Hz sampling frequency for 1 h daily over a period of 30 days. The detrended EN data were used to calculate the noise resistance (R _N) as well as the spectral noise resistance at zero frequency (R _SN^o) values and other statistical parameters. To study the nature of pits and distribution of their diameters as well as depths, extensive observations of the pitted and the blank specimens were carried out using Confocal Laser Scanning Microscopy (CLSM). The current and the potential records of the two alloys showed distinct differences in their corrosion behavior. It was observed that within less than 4 h of immersion, 316SS showed signals indicative of unstable pitting and onset of stable pitting was noticed after 48 h of exposure. However, until about 24 h, 316LN showed just the random signals and unstable pitting was observed after 28 h. The signals clearly indicated continuous growth of the stable pits in 316SS as against the repassivation of the unstable pits in 316LN after 7 days exposure. It was observed that R _N values showed a continuous decrease in the case of 316SS, but were quite stable in the case of 316LN over the exposure period. Concurrent to these observations it was observed that 316SS specimen was extensively pitted. The frequency distributions of pit diameters as well pit depths were observed to be highest at 10-20 μm and 5-10 μm, respectively. However, pits with as large as 70-80 μm diameter and as deep as 20-25 μm too were observed. No pits were observed in case of 316LN even after 30 days of exposure, an observation that corroborates well with the stable R _N values. Thus, in the present investigation, the long-term tests using EN technique coupled with CLSM studies conclusively prove that many unstable pits initiated in 316SS turn into stable ones resulting in insidious localized corrosion attack whereas the unstable pits initiated in 316LN get passivated in the simulated coastal environment.     

Grain growth kinetics in Dy-doped Bi0.5Na0.5TiO3 ceramics

Dysprosium-doped bismuth sodium titanate ceramics were prepared using the conventional mixed-oxide method. The amount of dysprosium used was varied from 0 to 2 at.%. The mixed powders were calcined at 800 °C and checked for phase purity using X-ray diffraction technique. The calcined powders were then cold-pressed into pellets and sintered at 1050 °C for the time ranging from 2 to 48 h. The ceramics were checked for phases and microstructures using an X-ray diffractometer and a scanning electron microscope, respectively. The analysis showed that undoped BNT ceramics sintered at longer time exhibited a significant grain growth with non-uniform grain size distribution and shape. The Dy-doped BNT however showed a much more limited grain growth behavior, resulting in smaller grain size and more equiaxed grain shape. It was also found that all Dy-doped BNT ceramics sintered at 48 h possessed lower porosity than those sintered for shorter time.     

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.     

Nucleation of branches in elastomeric polypropylene

The formation of a crosshatched morphology during crystallization of an isotactic polypropylene with low crystallinity is studied in detail. Here, a lamella seems to grow through another nearly perpendicular lamella that was crystallized before. The crystallization was observed with in situ scanning force microscopy with a temporal resolution of 3 min per image. After crystallization the volume structure was imaged by nanotomography. A detailed scheme for the nucleation of branches is proposed.