Mg-based implants have promising applications as biodegradable materials in medicine for orthopedic, dental, and cardiovascular therapies. During wear and degradation microdebris are released. Time-lapse multidimensional microscopy (MM) is proposed here as a suitable tool to follow, in fixed intervals over 24-h periods, the interaction between cells and particles. Results of MM show interactions of macrophages (J774) with the magnesium particles (MgPa) that led to modifications of cell size and morphology, a decrease in duplication rate, and cell damage. Corrosion products were progressively formed on the surface of the particles and turbulence was generated due to hydrogen development. Changes were more significant after treating MgPa with potassium fluoride. In order to complement MM observations, membrane damage as detected by a lactase dehydrogenase (LDH) assay and mitochondrial activity as detected by a WST-1 assay with macrophages and osteoblasts (MC3T3-E1) were compared. A more significant concentration-dependent effect was detected for macrophages exposed to MgPa than for osteoblasts. Accordingly, complementary data showed that viability and cell cycle seem to be more altered in macrophages. In addition, protein profiles and expression of proteins associated with the adhesion process changed in the presence of MgPa. These studies revealed that time-lapse MM is a helpful tool for monitoring changes of biodegradable materials and the biological surrounding in real time and in situ. This information is useful in studies related to biodegradable biomaterials.

In this work, fluorescent Bacillus subtilis (B. subtilis) cells were developed as probes for imaging applications and to explore behaviorial interaction between B. subtilis and Staphylococcus aureus (S. aureus). A novel biological strategy of coupling intracellular biochemical reactions for controllable biosynthesis of CdSe quantum dots by living B. subtilis cells was demonstrated, through which highly luminant and photostable fluorescent B. subtilis cells were achieved with good uniformity. With the help of the obtained fluorescent B. subtilis cells probes, S. aureus cells responded to co-cultured B. subtilis and to aggregate. The degree of aggregation was calculated and nonlinearly fitted to a polynomial model. Systematic investigations of their interactions implied that B. subtilis cells inhibit the growth of neighboring S. aureus cells, and this inhibition was affected by both the growth stage and the amount of surrounding B. subtilis cells. Compared to traditional methods of studying bacterial interaction between two species, such as solid culture medium colony observation and imaging mass spectrometry detection, the procedures were more simple, vivid, and photostable due to the efficient fluorescence intralabeling with less influence on the cells’ surface, which might provide a new paradigm for future visualization of microbial behavior.

The crayfish Cherax quadricarinatus stores calcium ions, easily mobilizable after molting, for calcifying parts of the new exoskeleton. They are chiefly stored as amorphous calcium carbonate (ACC) during each premolt in a pair of gastroliths synthesized in the stomach wall. How calcium carbonate is stabilized in the amorphous state in such a biocomposite remains speculative. The knowledge of the microstructure at the nanometer level obtained by field emission scanning electron microscopy and atomic force microscopy combined with scanning electron microscopy energy-dispersive X-ray spectroscopy, micro-Raman and X-ray absorption near edge structure spectroscopy gave relevant information on the elaboration of such an ACC-stabilized biomineral. We observed nanogranules distributed along chitin-protein fibers and the aggregation of granules in thin layers. AFM confirmed the nanolevel structure, showing granules probably surrounded by an organic layer and also revealing a second level of aggregation as described for other crystalline biominerals. Raman analyses showed the presence of ACC, amorphous calcium phosphate, and calcite. Elemental analyses confirmed the presence of elements like Fe, Na, Mg, P, and S. P and S are heterogeneously distributed. P is present in both the mineral and organic phases of gastroliths. S seems present as sulfate (probably as sulfated sugars), sulfonate, sulfite, and sulfoxide groups and, in a lesser extent, as sulfur-containing amino acids.

Conventional and highly sophisticated analytical methods (Cyria et al., 1989; Massar et al., 2012a) were used to analyze micro-structural and micro-analytical aspects of the blood of snake head fish, Channa gachua, exposed to municipal wastes and city garbage. Red (RBC) and white blood cell (WBC) counts and hemhemoglobin content were found to be higher in pollution affected fish as compared with control. Scanning electron microscopy revealed the occurrence of abnormal erythrocytes such as crenated cells, echinocytes, lobopodial projections, membrane internalization, spherocytes, ruptured cells, contracted cells, depression, and uneven elongation of erythrocyte membranes in fish inhabiting the polluted sites. Energy-dispersive X-ray spectroscopy (EDS) revealed the presence of silicon and lead in the RBCs of pollution affected fish. Significance of the study includes the highly sophisticated analytical approach, which revealed the aforementioned micro-structural abnormalities.

Millipedes are ecologically important soil organisms and may also be an economically threatening species in rural and urban areas when population outbreaks occur. In order to control infestations commercial formulations of deltamethrin have been commonly applied, even though there are few studies about the effects of such insecticide on millipedes. This paper describes the effects of this insecticide on millipedes showing neurotoxic effects assessed by synapsin labeling and confocal microscopy. Deltamethrin concentrations related to the DL50 of the active ingredient and a field concentration were applied topically in the diplopod Gymnostreptus olivaceus to evaluate the behavior, mortality rate, and synapsin levels in the brain 12, 24, and 48h after contact with deltamethin. The insecticide caused mortality at the higher concentrations employed, in which no change was observed in neurotransmission in the survivors. In contrast, at field concentrations, deltamethrin did not cause any deaths, but triggered significant changes in synapsin levels. The results obtained form the synapsin labeling provide several interpretations suggesting that the isolated application of this tool must be associated with additional tools in order to evaluate biologically induced effects of deltamethrin in an accurate way. In addition, the feasibility of chemical control of millipedes with deltamethrin is questioned.

Characterization of collagen fiber angle distribution throughout the blood vessel wall provides insight into the mechanical behavior of healthy and diseased arteries and their capacity to remodel. Atherosclerotic plaque contributes to the overall mechanical behavior, yet little is known experimentally about how collagen fiber orientation is influenced by atherogenesis. We hypothesized that atherosclerotic lesion development, and the factors contributing to lesion development, leads to a shift in collagen fiber angles within the aorta. Second-harmonic generation microscopy was used to visualize the three-dimensional organization of collagen throughout the aortic wall and to examine structural differences in mice maintained on high-fat Western diet versus age-matched chow diet mice in a model of atherosclerosis. Image analysis was performed on thoracic and abdominal sections of the aorta from each mouse to determine fiber orientation, with the circumferential (0°) and blood flow directions (axial ±90°) as the two reference points. All measurements were used in a multiple regression analysis to determine the factors having a significant influence on mean collagen fiber angle. We found that mean absolute angle of collagen fibers is 43° lower in Western diet mice compared with chow diet mice. Mice on a chow diet have a mean collagen fiber angle of ±63°, whereas mice on a Western diet have a more circumferential fiber orientation (~20°). This apparent shift in absolute angle coincides with the development of extensive aortic atherosclerosis, suggesting that atherosclerotic factors contribute to collagen fiber angle orientation.

An original high-pressure microscopy chamber has been designed for real-time visualization of biological cell growth during high isostatic (gas or liquid) pressure treatments up to 200 MPa. This new system is highly flexible allowing cell visualization under a wide range of pressure levels as the thickness and the material of the observation window can be easily adapted. Moreover, the design of the observation area allows different microscope objectives to be used as close as possible to the observation window. This chamber can also be temperature controlled. In this study, the resistance and optical properties of this new high-pressure chamber have been tested and characterized. The use of this new chamber was illustrated by a real-time study of the growth of two different yeast strains – Saccharomyces cerevisiae and Candida viswanathii – under high isostatic gas pressure (30 or 20 MPa, respectively). Using image analysis software, we determined the evolution of the area of colonies as a function of time, and thus calculated colony expansion rates.

The new generation of energy-dispersive X-ray (EDX) detectors with higher count rates than ever before, paves the way for a new approach to quantitative elemental analysis in the scanning transmission electron microscope. Here we demonstrate a method of calculating partial cross sections for use in quantifying EDX data, beneficial especially because of the simplicity of its implementation. Applying this approach to acid-leached PtCo catalyst nanoparticles leads to quantitative determination of the Pt surface enrichment.

Scanning electron microscope-cathodoluminescence (SEM-CL) analysis was performed for neodymium–iron–boron (NdFeB) and samarium–cobalt (Sm–Co) magnets to analyze the rare-earth elements present in the magnets. We examined the advantages of SEM-CL analysis over conventional analytical methods such as SEM-energy-dispersive X-ray (EDX) spectroscopy and SEM-wavelength-dispersive X-ray (WDX) spectroscopy for elemental analysis of rare-earth elements in NdFeB magnets. Luminescence spectra of chloride compounds of elements in the magnets were measured by the SEM-CL method. Chloride compounds were obtained by the dropwise addition of hydrochloric acid on the magnets followed by drying in vacuum. Neodymium, praseodymium, terbium, and dysprosium were separately detected in the NdFeB magnets, and samarium was detected in the Sm–Co magnet by the SEM-CL method. In contrast, it was difficult to distinguish terbium and dysprosium in the NdFeB magnet with a dysprosium concentration of 1.05 wt% by conventional SEM-EDX analysis. Terbium with a concentration of 0.02 wt% in an NdFeB magnet was detected by SEM-CL analysis, but not by conventional SEM-WDX analysis. SEM-CL analysis is advantageous over conventional SEM-EDX and SEM-WDX analyses for detecting trace rare-earth elements in NdFeB magnets, particularly dysprosium and terbium.

Micro-scale observations in karst caves help to identify different processes that shaped local morphology. Scanning electron microscopy/energy-dispersive X-ray spectroscopy inspection of speleothems from two karst caves in Slovenia, Predjama and Črna Jama, confirmed the presence of sub-angular to sub-rounded detrital fragments of clay minerals, feldspars, quartz, Fe-oxides/hydroxides, rutile and Nb-rutile, xenotime, kassite, allanite, fluorapatite, epidote, ilmenite, monazite, sphene, and zircon, between 2 and 50 μm across. These occur in porous layers separating calcite laminae in the clayey coating on the layer below the surface of the speleothems, and are also incorporated within actual crystals. It is likely that they are derived from the weathered rocks of the Eocene flysch. Probably they were first transported into the caves by floodwaters forming cave sediments. Later, depending upon the climate conditions, they were moved by air currents or by water to the surface of active speleothems. They might also be redeposited from overlying soils enriched with wind-transported minerals from the flysch, or from higher passages filled with weathered flysch sediment, by drip water percolating through the fissured limestone. As some of the identified minerals are carriers of rare earth elements, Ti and Zr, their presence could affect any palaeoclimatic interpretations that are based upon the geochemical composition of the speleothems.

Phase identification of multi-phase materials provides essential information relating the material to its mechanical properties. In this study we selected DP980, a type of dual-phase steel, to investigate the content of martensite and ferrite grains. A combination of advanced techniques was used to provide detailed and precise information of the microstructure. Scanning and transmission electron microscopy were used to provide observations of the sample surface at different scales. Martensite and ferrite phases of DP980 were further identified and characterized using electron backscatter diffraction and scanning probe microscopy. Results obtained with nanoindentation tests confirmed that the differences in nanohardness values in single-phase grains are martensite and ferrite with different surface heights shown by scanning probe microscopy. The similarity shown in the image quality map and scanning probe microscopy proves that a large fraction of martensite can be distinguished in this undeformed material using image quality parameters obtained during electron backscatter diffraction imaging.

In this study low temperature micro-photoluminescence technology was employed to investigate effects of the irradiation and nitrogen concentration on nitrogen-vacancy (NV) luminescence, with the photochromic and vibronic properties of the NV defects. Results showed that the NV luminescence was weakened due to recombination of self-interstitials created by electron irradiation in diamond and the vacancies within the structure of NV centers. For very pure diamond, the vacancies migrated the long distance to get trapped by N atoms only after sufficient high temperature annealing. As with the increase in nitrogen content, the migration distance of vacancies got smaller. The nitrogen also favored the formation of negatively charged NV centers with the donating electrons. Under the high-energy ultraviolet laser excitation, the photochromic property of the NV− center was also observed, though it was not stable. Besides, the NV centers showed very strong broad sidebands, and the vibrations involved one phonon with energy of ~42 meV and another with ~67 meV energy.

(Ba0.5Sr0.5)(Co0.8Fe0.2)O3−δ (BSCF) is a promising material with mixed ionic and electronic conductivity which is considered for oxygen separation membranes. Selective improvement of material properties, e.g. oxygen diffusivity or suppression of secondary phase formation, can be achieved by B-site doping. This study is concerned with the formation of Co-oxide precipitates in undoped BSCF at typical homogenization temperatures of 1,000°C, which act as undesirable nucleation sites for other secondary phases in the application-relevant temperature range. Y-doping successfully suppresses Co-oxide formation, whereas only minor improvements are achieved by Sc-doping. To understand the reason for the different behavior of Y and Sc, the lattice sites of dopant cations in BSCF were experimentally determined in this work. Energy-dispersive X-ray spectroscopy in a transmission electron microscope was applied to locate dopant sites exploiting the atom location by channeling enhanced microanalysis technique. It is shown that Sc exclusively occupies B-cation sites, whereas Y is detected on A- and B-cation sites in Y-doped BSCF, although solely B-site doping was intended. A model is presented for the suppression of Co-oxide formation in Y-doped BSCF based on Y double-site occupancy.

A novel type of alumina (Al2O3)-doped molybdenum (Mo) alloy sheet was prepared by a hydrothermal method and a subsequent powder metallurgy process. Then the characterization of α-Al2O3 was investigated using high-resolution transmission electron microscopy as the research focus. The tensile strength of the Al2O3-doped Mo sheet is 43–85% higher than that of the pure Mo sheet, a very obvious reinforcement effect. The sub-micron and nanometer-scale Al2O3 particles can increase the recrystallization temperature by hindering grain boundary migration and improve the tensile strength by effectively blocking the motion of the dislocations. The Al2O3 particles have a good bond with the Mo matrix and there exists an amorphous transition layer at the interface between Al2O3 particles and the Mo matrix in the as-rolled sheet. The sub-structure of α-Al2O3 is characterized by a number of nanograins in the $\left[ {2\bar{2}1} \right]$ direction. Lastly, a new computer-based method for indexing diffraction patterns of the hexagonal system is introduced, with 16 types of diffraction patterns of α-Al2O3 indexed.

Melt-back etching is an effect that can occur for gallium (Ga) containing III/V semiconductors grown on Si. Since this effect influences interfaces between the two compounds and therefore the physical characteristics of the material composition, it is desirable to understand its driving forces. Therefore, we investigated Ga grown on Si (001) via metal organic chemical vapor deposition using trimethyl Ga as a precursor. As a result of the melt-back etching, Ga-containing droplets formed on the Si surface which reach into the Si wafer. The shape of these structures was analyzed by plan view investigation and cross sectional tomography in a (scanning) transmission electron microscope. For plan view preparation a focused ion beam was used to avoid damage to the Ga-containing structures, which are sensitive to the chemicals normally used during conventional plan view preparation. Combining the results of both investigation methods confirms that the Ga-containing structure within the Si exhibits a pyramid shape with facets along the Si {111} lattice planes.

Three-phase three-dimensional (3D) microstructural reconstructions of lithium-ion battery electrodes are critical input for 3D simulations of electrode lithiation/delithiation, which provide a detailed understanding of battery operation. In this report, 3D images of a LiCoO2 electrode are achieved using focused ion beam-scanning electron microscopy (FIB-SEM), with clear contrast among the three phases: LiCoO2 particles, carbonaceous phases (carbon and binder) and the electrolyte space. The good contrast was achieved by utilizing an improved FIB-SEM sample preparation method that combined infiltration of the electrolyte space with a low-viscosity silicone resin and triple ion-beam polishing. Morphological parameters quantified include phase volume fraction, surface area, feature size distribution, connectivity, and tortuosity. Electrolyte tortuosity was determined using two different geometric calculations that were in good agreement. The electrolyte tortuosity distribution versus position within the electrode was found to be highly inhomogeneous; this will lead to inhomogeneous electrode lithiation/delithiation at high C-rates that could potentially cause battery degradation.

This paper intends to analyze the microstructure of concrete with recycled aggregates (RA) from construction and demolition waste from various Portuguese recycling plants. To that effect, several scanning electron microscopy (SEM) and energy-dispersive X-ray spectroscopy (EDS) analyses were performed. Various concrete mixes were evaluated in order to analyze the influence of the RA’s collection point and consequently of their composition on the mixes’ characteristics. Afterward all the mixes were subjected to the capillary water absorption test in order to quantitatively evaluate their porosity. Results from the SEM/EDS analysis were compared with those from capillary water absorption test. The SEM/EDS analysis showed that the bond capacity of aggregates to the new cement paste is greatly influenced by the RA’s nature. On the other hand, there was an increase in porosity with the incorporation of RA.

Biomimetic hierarchical surface structures that exhibit features having multiple length scales have been used in many technological and engineering applications. Their surface topographies are most commonly analyzed using scanning electron microscopy (SEM), which only allows for qualitative visual assessments. Here we introduce fractal and lacunarity analyses as a method of characterizing the SEM images of hierarchical surface structures in a quantitative manner. Taking femtosecond laser-irradiated metals as an example, our results illustrate that, while the fractal dimension is a poor descriptor of surface complexity, lacunarity analysis can successfully quantify the spatial texture of an SEM image; this, in turn, provides a convenient means of reporting changes in surface topography with respect to changes in processing parameters. Furthermore, lacunarity plots are shown to be sensitive to the different length scales present within a hierarchical structure due to the reversal of lacunarity trends at specific magnifications where new features become resolvable. Finally, we have established a consistent method of detecting pattern sizes in an image from the oscillation of lacunarity plots. Therefore, we promote the adoption of lacunarity analysis as a powerful tool for quantitative characterization of, but not limited to, multi-scale hierarchical surface topographies.

A new closed cell is presented for in situ X-ray ptychography which allows studies under gas flow and at elevated temperature. In order to gain complementary information by transmission and scanning electron microscopy, the cell makes use of a Protochips E-chipTM which contains a small, thin electron transparent window and allows heating. Two gold-based systems, 50 nm gold particles and nanoporous gold as a relevant catalyst sample, were used for studying the feasibility of the cell. Measurements showing a resolution around 40 nm have been achieved under a flow of synthetic air and during heating up to temperatures of 933 K. An elevated temperature exhibited little influence on image quality and resolution. With this study, the potential of in situ hard X-ray ptychography for investigating annealing processes of real catalyst samples is demonstrated. Furthermore, the possibility to use the same sample holder for ex situ electron microscopy before and after the in situ study underlines the unique possibilities available with this combination of electron microscopy and X-ray microscopy on the same sample.

Different quartz types from several localities in the Czech Republic and Sweden were examined by polarizing microscopy combined with cathodoluminescence (CL) microscopy, spectroscopy, and petrographic image analysis, and tested by use of an accelerated mortar bar test (following ASTM C1260). The highest alkali–silica reaction potential was indicated by very fine-grained chert, containing significant amounts of fine-grained to cryptocrystalline matrix. The chert exhibited a dark red CL emission band at ~640 nm with a low intensity. Fine-grained orthoquartzites, as well as fine-grained metamorphic vein quartz, separated from phyllite exhibited medium expansion values. The orthoquartzites showed various CL of quartz grains, from blue through violet, red, and brown. Two CL spectral bands at ~450 and ~630 nm, with various intensities, were detected. The quartz from phyllite displayed an inhomogeneous dark red CL with two CL spectral bands of low intensities at ~460 and ~640 nm. The massive coarse-grained pegmatite quartz from pegmatite was assessed to be nonreactive and displayed a typical short-lived blue CL (~480 nm). The higher reactivity of the fine-grained hydrothermal quartz may be connected with high concentrations of defect centers, and probably with amorphized micro-regions in the quartz, respectively; indicated by a yellow CL emission (~570 nm).