Scale-bridging analysis on deformation behavior of high-nitrogen austenitic Fe–18Cr–10Mn–(0.39 and 0.69)N steels was performed by neutron diffraction, electron backscattered diffraction (EBSD), and transmission electron microscopy (TEM). Two important modes of deformation were identified depending on the nitrogen content: deformation twinning in the 0.69 N alloy and strain-induced martensitic transformation in the 0.39 N alloy. The phase fraction and deformation faulting probabilities were evaluated based on analyses of peak shift and asymmetry of neutron diffraction profiles. Semi in situ EBSD measurement was performed to investigate the orientation dependence of deformation microstructure and it showed that the variants of ε martensite as well as twin showed strong orientation dependence with respect to tensile axis. TEM observation showed that deformation twin with a {111}⟨112⟩ crystallographic component was predominant in the 0.69 N alloy whereas two types of strain-induced martensites (ε and α′ martensites) were observed in the 0.39 N alloy. It can be concluded that scale-bridging analysis using neutron diffraction, EBSD, and TEM can yield a comprehensive understanding of the deformation mechanism of nitrogen-alloyed austenitic steels.

Ferruginous bodies observed in lungs of patients affected by mesothelioma, asbestosis, and pulmonary carcinoma are important to relate the illness to exposure, environmental or occupational, to asbestos. Identification of the inorganic phase constituting the core of the ferruginous bodies, formed around asbestos but also around phases different from asbestos, is essential for legal purposes. Environmental scanning electron microscopy/energy dispersive spectroscopy was used to identify the fibrous mineral phase in the core of ferruginous bodies observed directly in thin sections of tissue, without digestion of the biological matrix. Spectra were taken with sequential analyses along a line crossing the core of the ferruginous bodies. By comparing the spectra taken near to and far from the core, the chemical elements that make up the core could be identified.

High-resolution characterizations of intergranular attack in alloy 600 (Ni-17Cr-9Fe) exposed to 325°C simulated pressurized water reactor primary water have been conducted using a combination of scanning electron microscopy, NanoSIMS, analytical transmission electron microscopy, and atom probe tomography. The intergranular attack exhibited a two-stage microstructure that consisted of continuous corrosion/oxidation to a depth of ~200 nm from the surface followed by discrete Cr-rich sulfides to a further depth of ~500 nm. The continuous oxidation region contained primarily nanocrystalline MO-structure oxide particles and ended at Ni-rich, Cr-depleted grain boundaries with spaced CrS precipitates. Three-dimensional characterization of the sulfidized region using site-specific atom probe tomography revealed extraordinary grain boundary composition changes, including total depletion of Cr across a several nm wide dealloyed zone as a result of grain boundary migration.

Direct imaging of nanoscale objects suspended in liquid media can be accomplished using cryogenic transmission electron microscopy (cryo-TEM). Cryo-TEM has been used with particular success in microbiology and other biological fields. Samples are prepared by plunging a thin film of sample into an appropriate cryogen, which essentially produces a snapshot of the suspended objects in their liquid medium. With successful sample preparation, cryo-TEM images can facilitate elucidation of aggregation and self-assembly, as well as provide detailed information about cells and viruses. This work provides an explanation of sample preparation, detailed examples of the many artifacts found in cryo-TEM of aqueous samples, and other key considerations for successful cryo-TEM imaging.

A mean-shift clustering (MSC) algorithm is introduced as a valuable alternative to perform materials phase classification from multispectral images. As opposed to other multivariate statistical techniques, such as factor analysis or principal component analysis (PCA), clustering techniques directly assign a class label to each pixel, so that their outputs are phase segmented images, i.e., there is no need for an additional segmentation algorithm. On the other hand, as compared to other clustering procedures and classification methods, such as segmentation by thresholding of multiple spectral components, MSC has the advantages of not requiring previous knowledge of the number of data clusters and not assuming any shape for these clusters, i.e., neither the number nor the composition of the phases must be previously known. This makes MSC a particularly useful tool for exploratory research, assisting phase identification of unknown samples. Visualization and interpretation of the results are also simplified, since the information content of the output image does not depend on the particular choice of the content of the color channels. We applied MSC to the analysis of two sets of X-ray maps acquired in scanning electron microscopes equipped with energy-dispersive detection systems. Our results indicate that MSC is capable of detecting additional phases, not clearly identified through PCA or multiple thresholding, with a very low empirical reject rate.

It is well known that extracellular signal-regulated kinase 8 (ERK8) plays pivotal roles in various mitotic events. But its physiological roles in oocyte meiotic maturation remain unclear. In this study, we found that although no specific ERK8 signal was detected in oocyte at the germinal vesicle stage, ERK8 began to migrate to the periphery of chromosomes shortly after germinal vesicle breakdown. At prometaphase I, metaphase I (MI), anaphase I, telophase I, and metaphase II (MII) stages, ERK8 was stably detected at the spindles. By taxol treatment, we clarified that the ERK8 signal was stained on the spindle fibers as well as microtubule asters in MI and MII oocytes. In fertilized eggs, the ERK8 signal was not observed in the two pronuclei stages. At prometaphase, metaphase, and anaphase of the first mitosis, ERK8 was detected on the mitotic spindle. ERK8 knock down by antibody microinjection and specific siRNA caused abnormal spindles, failed chromosome congression, and decreased first polar body extrusion. Taken together, our results suggest that ERK8 plays an important role in spindle organization during mouse oocyte meiotic maturation and early embryo cleavage.

Methods to determine the rotation axis using the rotation electron diffraction technique are described. A combination of rotation axis tilt, beam tilt, and simulated experimental diffraction patterns with nonintegers zone axis has been used. Accurate knowledge of the crystallographic direction of the incident beam for deducing the excitation error of reflections simultaneously near Bragg positions is essential in quantitative electron diffraction. Experimental patterns from CoP3 are used as examples.

Hepatolithiasis (HL) or intrahepatic calculi is an uncommon condition in Western countries, with a prevalence under 1%. It is much more frequent in East Asia, reaching 20% in China and Taiwan, up to 50% of which show associated cholelithiasis. The authors report a European patient with primary HL of the left hepatic lobe, 12 years after cholecystectomy for acute calculous cholecystitis, now successfully treated after left lobe hepatectomy.

A 63-year-old caucasian man with a past medical history of hypertension, coronary artery disease, dyslipidaemia and urgent cholecystectomy 12 years ago (due to an acute calculous cholecystitis without ultrasonographic or perioperative evidence of bile duct dilation or choledocholithiasis, and with no cholangitis ever since) was now admitted to our ward following a 2-day course of fever (39ºC), epigastric pain, nausea, vomiting and tender right upper abdominal quadrant.

Bloodwork showed mild leukocytosis with neutrophilia and low platelets; elevated C-reactive protein, γGT, alkaline phosphatase and LDH; normal AST, ALT, bilirubin and amylase. Negative HBV, HCV and HIV serologies. CA19.9 was notably high (7500 U/mL), 200x above normal range (N≤37). A diagnosis of acute cholangitis was assumed on clinical and laboratory basis, despite the absence of jaundice. The patient began therapy with iv ceftriaxone and improved clearly over the next few days. Raised CA19.9 in the setting of acute cholangitis, however, forced us into further study directed at the possibility of underlying biliary or pancreatic malignancy.

Abdominal ultrasound disclosed multiple calcifications in the left lobe. CT displayed several left lobe intrahepatic bile duct dilations but no calcifications, thus suggesting intrahepatic cholesterol stones and cholangitis due to an obstacle before the hilum. MR-cholangiography was performed, showing marked dilation of the left intrahepatic bile duct and moderate dilation in the extrahepatic portion of the common bile duct (CBD). Neither exam showed any sign of cancer. Surgery was the therapeutic approach, and the patient underwent a left hepatectomy.

Intra-operative ultrasound study unveiled severe dilation of the left intrahepatic bile duct, which was filled with gallstones, but no choledocholithiasis and no tumour whatsoever. Both the surgical procedure and the post-operative period were uneventful.

Macroscopic liver section showed multiple dilated bile ductules containing fragile yellowish-green gallstones (Fig.1). Microscopy revealed cystic dilations of the left intrahepatic biliary tree packed with intraluminal gallstones (Fig.2), fibrosis and moderate chronic inflammatory infiltrate with lymphoid aggregates (Fig.3). Some areas of the epithelium displayed erosion and reactive changes. No biliary neoplasm was found.

Immunocytochemistry for angiogenesis or lymphocyte membrane markers are not available in our Hospital. Soon after surgery, CA19.9 values decreased abruptly and just 4 months after the left hepatectomy they were back within reference range.

Our working hypotheses were primary hepatolithiasis (HL), secondary HL and Caroli disease.

Secondary HL, due to stone migration from the CBD, was discarded because calculi in the CBD were absent in the cholecystectomy 12 years ago and in ultrasonographic studies both then and now. Caroli disease, a congenital condition characterised by intrahepatic bile duct dilation, was contradicted by the ultrasonography in 2000 showing normal bile duct dimension. Excluding these two entities made primary idiopathic HL our final diagnosis.

The patient is now asymptomatic, having returned to his normal life with no limitations.

This report showcases a rare entity known as primary hepatolithiasis, usually causing recurrent cholangitis in older patients of Asian descent but seldom seen in Europe. The steep increase in CA19.9 related to cholangitis, although previously reported, is also very uncommon. Another interesting aspect is the conspicuous inexistence, over 12 years, of recurrent episodes of acute cholangitis (a traditional finding in hepatolithiasis). These three features help to compose this most peculiar case.

Band contrast (BC) is a qualitative measure of electron back-scattered diffraction (EBSD), which is derived from the intensity of the Kikuchi bands. The BC is dependent upon several factors including scanning electron microscope measurement parameters, EBSD camera setup, and the specimen itself (lattice defect and grain orientation). In this study, the effective factors for BC variations and the feasibility of using BC variations for the quantification of microstructure evolutions have been investigated. In addition, the effects of the lattice defect and the grain orientation on the BC variations are studied. Next, a shear-deformed microstructure of 316L stainless steel, which contains nanosized grains and a large portion of twin boundaries, is revealed by BC map and histogram. Recovery and recrystallization of shear-deformed 316L stainless steel are displayed by BC variations during isothermal annealing at 700 and 800°C, respectively. It is observed that the BC turns bright as the shear-deformed crystal structure is recovered or recrystallized.

Extended abstract of a paper presented at Microscopy and Microanalysis 2013 in Indianapolis, Indiana, USA, August 4 – August 8, 2013.

The BiMeVOx family of compounds appears to be more attractive for applications at low temperatures when ionic conductivity is the determining parameter. The objective of this study was to analysis the influence of voltage of the behavior of the Schottky barrier in both BiCuVOX and BiTiVOX. The samples were analyzed by atomic force microscopy and electric force microscopy (EFM). EFM experiments were conducted to map the electric field distribution on the surface. The formation of Schottky barriers was observed, and their height and width measured. BiCuVOX samples show a barrier width of 140 nm, and BiTiVOX shows a barrier width of 350 nm. The applied voltage has no effect on the barrier width but increases the peak height as observed in the cantilever frequency as measured with the EFM technique.

Ionic liquids (ILs) and their mixtures with low molecular solvents present ideal properties for use as flotation liquids in cryo-ultramicrotomy. With control of Tg and η by co-solvent addition, flat, ultra-thin sections are reliably floated onto transmission electron microscopy grids even at temperatures as low as −100°C. Even more, the liquids and their mixtures are stable in the microtome trough for several hours because of low vapor pressure and low solidification temperature. Compared to established flotation media for soft polymer systems, the time and skill needed for cryo-ultramicrotomy are significantly reduced. Although just a handful of ILs are discussed and a good general choice identified, if different liquid characteristics are needed for a particular sample, thousands of additional ILs will perform similarly, giving this new approach enormous flexibility.

The advent of simultaneous energy dispersive X-ray spectroscopy (EDS) data collection has vastly improved the phase separation capabilities for electron backscatter diffraction (EBSD) mapping. A major problem remains, however, in distinguishing between multiple cubic phases in a specimen, especially when the compositions of the phases are similar or their particle sizes are small, because the EDS interaction volume is much larger than that of EBSD and the EDS spectra collected during spatial mapping are generally noisy due to time limitations and the need to minimize sample drift. The backscatter electron (BSE) signal is very sensitive to the local composition due to its atomic number (Z) dependence. BSE imaging is investigated as a complimentary tool to EDS to assist phase segmentation and identification in EBSD through examination of specimens of meteorite, Cu dross, and steel oxidation layers. The results demonstrate that the simultaneous acquisition of EBSD patterns, EDS spectra, and the BSE signal can provide new potential for advancing multiphase material characterization in the scanning electron microscope.

We propose a nonscanning three-dimensional (3D) optical microscope based on reflectivity-height transformation in applications of biological and transparent plate measurements. The reflectivity of a prism can be transformed to the surface height of the specimen based on geometrical optics and the principle of internal reflection. Thus, the pattern of reflectivity is representative of the surface profile. Using charge-coupled device cameras to obtain the two-dimensional image patterns and combining with its reflectivity pattern, the 3D profile can be generated. The lateral resolution is determined by the diffraction limit, and the vertical resolution is better than several nanometers according to the incident angle and polarization used.

With the advent of in vivo laser scanning fluorescence microscopy techniques, time-series and three-dimensional volumes of living tissue and vessels at micron scales can be acquired to firmly analyze vessel architecture and blood flow. Analysis of a large number of image stacks to extract architecture and track blood flow manually is cumbersome and prone to observer bias. Thus, an automated framework to accomplish these analytical tasks is imperative. The first initiative toward such a framework is to compensate for motion artifacts manifest in these microscopy images. Motion artifacts in in vivo microscopy images are caused by respiratory motion, heart beats, and other motions from the specimen. Consequently, the amount of motion present in these images can be large and hinders further analysis of these images. In this article, an algorithmic framework for the correction of time-series images is presented. The automated algorithm is comprised of a rigid and a nonrigid registration step based on shape contexts. The framework performs considerably well on time-series image sequences of the islets of Langerhans and provides for the pivotal step of motion correction in the further automatic analysis of microscopy images.

The understanding of solid–gas interactions has been greatly advanced over the past decade on account of the availability of high-resolution transmission electron microscopes (TEMs) equipped with differentially pumped environmental cells. The operational pressures in these differentially pumped environmental TEM (DP-ETEM) instruments are generally limited up to 20 mbar. Yet, many industrial catalytic reactions are operated at pressures equal or higher than 1 bar—50 times higher than that in the DP-ETEM. This poses limitations for in situ study of gas reactions through ETEM and advances are needed to extend in situ TEM study of gas reactions to the higher pressure range. Here, we present a first series of experiments using a gas flow membrane cell TEM holder that allows a pressure up to 4 bar. The built-in membrane heaters enable reactions at a temperature of 95–400°C with flowing reactive gases. We demonstrate that, using a conventional thermionic TEM, 2 Å atomic fringes can be resolved with the presence of 1 bar O2 gases in an environmental cell and we show real-time observation of the Kirkendall effect during oxidation of cobalt nanocatalysts.

A new automated crystallographic orientation mapping tool in a transmission electron microscope technique, which is based on pattern matching between every acquired electron diffraction pattern and precalculated templates, has been used for the microstructural characterization of nondeformed and deformed aluminum thin films and twinning-induced plasticity steels. The increased spatial resolution and the use of electron diffraction patterns rather than Kikuchi lines make this tool very appropriate to characterize fine grained and deformed microstructures.

We investigated an eclogitic gneiss from the Western Gneiss Region in Norway, which underwent subduction as part of Baltica lithosphere beneath Laurentia during the Scandian orogeny. Petrologic data indicate that the eclogite was deformed plastically at about 4 GPa and 800°C producing a strong macroscopic foliation and lineation. Whereas garnet remained largely stable during the retrograde uplift, omphacite was transformed statically into a symplectite consisting of lamellar diopside and plagioclase with more equant grains of hornblende and orthopyroxene. Measurements of the crystallographic preferred orientation with electron backscatter diffraction show that diopside and hornblende, as well as orthopyroxene, have a systematic orientation relationship with the macroscopic fabric, as well as the (presumed) orientation of the host omphacite. The orientation relationship between the chain silicates is very sharp with the crystallographic forms {100}, {010}, and ⟨001⟩ being parallel. Their bulk texture shows a maximum of ⟨001⟩ parallel to the lineation and girdles of {010} and {110} perpendicular to the lineation with maxima subparallel to the foliation corresponding to an L-type texture of the original omphacite and indicating constrictional strain with an additional component of pure shear/simple shear component.