Atom probe tomography is routinely used for the characterization of materials microstructures, usually assuming that the microstructure is unaltered by the analysis. When analyzing ionic conductors, however, gradients in the chemical potential and the electric field penetrating dielectric atom probe specimens can cause significant ionic mobility. Although ionic mobility is undesirable when aiming for materials characterization, it offers a strategy to manipulate materials directly in situ in the atom probe. Here, we present experimental results on the analysis of the ionic conductor lithium-manganese-oxide with different atom probe techniques. We demonstrate that, at a temperature of 30 K, characterization of the materials microstructure is possible without measurable Li mobility. Also, we show that at 298 K the material can be deintercalated, in situ in the atom probe, without changing the manganese-oxide host structure. Combining in situ atom probe deintercalation and subsequent conventional characterization, we demonstrate a new methodological approach to study ionic conductors even in early stages of deintercalation.

The choice of materials that constitute electrodes and the way they are interconnected, i.e., the microstructure, influences the performance of lithium-ion batteries. For batteries with high energy and power densities, the microstructure of the electrodes must be controlled during their manufacturing process. Moreover, understanding the microstructure helps in designing a high-performance, yet low-cost battery. In this study, we propose a systematic algorithm workflow for the images of the microstructure of anodes obtained from a focused ion beam scanning electron microscope (FIB-SEM). Here, we discuss the typical issues that arise in the raw FIB-SEM images and the corresponding preprocessing methods that resolve them. Next, we propose a Fourier transform-based filter that effectively reduces curtain artifacts. Also, we propose a simple, yet an effective, global-thresholding method to identify active materials and pores in the microstructure. Finally, we reconstruct the three-dimensional structures by concatenating the segmented images. The whole algorithm workflow used in this study is not fully automated and requires user interactions such as choosing the values of parameters and removing shine-through artifacts manually. However, it should be emphasized that the proposed global-thresholding method is deterministic and stable, which results in high segmentation performance for all sectioning images.

Acquiring and processing phase-contrast microscopy images in wide-field long-term live-cell imaging and high-throughput screening applications is still a challenge as the methodology and algorithms used must be fast, simple to use and tune, and as minimally intrusive as possible. In this paper, we developed a simple and fast algorithm to compute the cell-covered surface (degree of confluence) in phase-contrast microscopy images. This segmentation algorithm is based on a range filter of a specified size, a minimum range threshold, and a minimum object size threshold. These parameters were adjusted in order to maximize the F-measure function on a calibration set of 200 hand-segmented images, and its performance was compared with other algorithms proposed in the literature. A set of one million images from 37 myoblast cell cultures under different conditions were processed to obtain their cell-covered surface against time. The data were used to fit exponential and logistic models, and the analysis showed a linear relationship between the kinetic parameters and passage number and highlighted the effect of culture medium quality on cell growth kinetics. This algorithm could be used for real-time monitoring of cell cultures and for high-throughput screening experiments upon adequate tuning.

We evaluated the depth resolution of annular dark-field (ADF) scanning confocal electron microscopy (SCEM) with a stage-scanning system by observation of nanoparticles. ADF-SCEM is a three-dimensional (3D) imaging technique that we recently proposed. An ADF-SCEM instrument involves a pinhole aperture before a detector for rejecting electrons from the out-of-focal plane in a specimen and an annular aperture under the specimen for collecting only scattered electrons. The stage-scanning system enables us to directly obtain optical slice images perpendicular and parallel to an optical axis at a desired position. In particular, the parallel slices visualize the elongation of nanoparticles along the optical axis, which depends on the depth resolution. ADF-SCEM effectively reduced the elongation length of the nanoparticles sufficiently to demonstrate depth sectioning, in comparison with scanning transmission electron microscopy and bright-field SCEM. The experimentally obtained length was nearly equal to the theoretically estimated one from the probe size considering the experimental conditions. Furthermore, we applied this ADF-SCEM technique to analysis of the 3D position of catalytic nanoparticles on carbon nanostructures.

Environmental transmission electron microscopy (ETEM) is opening an important window for in situ studies of interaction of water with oxides. Studies of MgO smoke nanocrystals under partial pressures of water ranging from 10 mTorr to 10 Torr found their {100} neutral surfaces to be extremely resistant to dissociative adsorption of water and hydroxylation, in agreement with recent theoretical predictions. ETEM observations of electron irradiation driven MgO smoke nanocrystal hydroxylation displayed the anticipated volume expansion, but revealed complex shape changes with elongations toward oxide corners. The reaction rate was found to increase with electron flux at constant water pressure. In situ selected area diffraction studies of MgO single crystals showed that the hydroxide grows with its basal (0001) plane parallel to the polar MgO (111) planes. This is the same crystallographic relationship as in dehydroxylation experiments, but with four variants. Electron energy loss spectroscopy found oxygen K-edge changes consistent with bulk hydroxylation.

The current hypothesis of alveolar capillary membrane dysfunction fails to completely explain the severe and persistent leak of protein-rich fluid into the pulmonary interstitium, seen in the exudative phase of acute lung injury (ALI). The presence of intact red blood cells in the pulmonary interstitium may suggest mechanical failure of pulmonary arterioles and venules. These studies involved the pathological and ultrastructural evaluation of the pulmonary vasculature in Staphylococcal enterotoxin B (SEB)-induced ALI. Administration of SEB resulted in a significant increase in the protein concentration of bronchoalveolar lavage fluid and vascular leak in SEB-exposed mice compared to vehicle-treated mice. In vivo imaging of mice demonstrated the pulmonary edema and leakage in the lungs of SEB-administered mice. The histopathological studies showed intense clustering of inflammatory cells around the alveolar capillaries with subtle changes in architecture. Electron microscopy studies further confirmed the diffuse damage and disruption in the muscularis layer of the terminal vessels. Cell death in the endothelial cells of the terminal vessels was confirmed with TUNEL staining. In this study, we demonstrated that in addition to failure of the alveolar capillary membrane, disruption of the pulmonary arterioles and venules may explain the persistent and severe interstitial and alveolar edema.

One of the critical prerequisites for accurately measuring the mechanical properties of profiled fibers is the precise determination of their cross-sectional areas (CSAs). In this study, a new method is established for determining a single profiled fibers’ CSA based on the frozen section method and digital photo, pixel-ratio method (FS-DP). FS-DP is used to obtain a transverse section of a fiber, by acquiring an image of the cross section using optical microscopy or scanning electron microscopy, and then calculating the CSA using Photoshop. Using FS-DP, it was found that the shape of a fiber of silk changes little in a range of 50 μm, but varies considerably over a range of 1 m, while the CSA of cocoon silk (900 m) first increases and then decreases. Mechanical property tests showed that the elongation, strength, elastic modulus, and toughness values of the cocoon silk are consistent with those reported previously. Additionally, FS-DP was also used to observe other profiled fibers. The application tests indicated that FS-DP can be used to quickly and accurately obtain the CSA of a single profiled fiber, and that it is suitable for the large-scale determination and analysis of the mechanical properties of profiled fibers.

The emergence of commercial electron backscatter diffraction (EBSD) equipment ushered in an era of information rich maps produced by determining the orientation of user-selected crystal structures. Since then, a technological revolution has occurred in the quality, rate detection, and analysis of these diffractions patterns. The next revolution in EBSD is the ability to directly utilize the information rich diffraction patterns in a high-throughput manner. Aided by machine learning techniques, this new methodology is, as demonstrated herein, capable of accurately separating phases in a material by crystal symmetry, chemistry, and even lattice parameters with fewer human decisions. This work is the first demonstration of such capabilities and addresses many of the major challenges faced in modern EBSD. Diffraction patterns are collected from a variety of samples, and a convolutional neural network, a type of machine learning algorithm, is trained to autonomously recognize the subtle differences in the diffraction patterns and output phase maps of the material. This study offers a path to machine learning coupled phase mapping as databases of EBSD patterns encompass an increasing number of the possible space groups, chemistry changes, and lattice parameter variations.

Transmission electron microscopy characterization of two major long-period stacking ordered (LPSO) phases in Mg–Zn–Y alloy, i.e., 18R- and 14H-LPSO are reported. The space group and atomic-scale microstructures of both compounds were determined using a combination of electron diffraction, convergent beam electron diffraction, high-resolution transmission electron microscopy, and Z-contrast scanning transmission electron microscopy. The 18R-LPSO phase is demonstrated to have a point group and space group 3m and R3m (or 3m and R3m), with the lattice parameter a = 1.112 nm and c = 4.689 nm in a hexagonal coordinate system. The 14H-LPSO phase has a point group 6/mmm and a space group P63 /mmc, and the lattice parameter is a = 1.112 nm and c = 3.647 nm. In addition, insertion of extra thin Mg platelets of several atomic layers, results in stacking faults in the LPSO phase. These results may shed some new light on a better understanding of the microstructure and deformation mechanisms of LPSO phases in Mg alloys.

In this paper we describe the preparation of fluorescent mesoporous silica nanoparticles (MSNs) for traceable drug delivery systems. The nanoparticles were prepared following a sol–gel procedure, incorporating a modified perylenediimide dye in the silica structure. Transmission electron microscopy and scanning electron microscopy show that the nanoparticles are monodispersed, with a spheroid shape and a raspberry-type surface morphology. The hybrid MSNs are robust, maintaining the mesoporous structure after template removal, with a pore diameter above 2 nm. A polymer shell was synthesized from the external surface of the hybrid nanoparticles by atom transfer radical polymerization, showing temperature-switchable collapsed/expanded conformation control. The fluorescent properties of the perylenediimide dye incorporated in the MSN pore walls are intact, and internalization in HEK293 cells shows that the nanoparticles are efficiently dispersed in the cytosol. These results show that the mesoporous fluorescent hybrid nanoparticles are an excellent platform for development of a traceable drug delivery system.

Cemented tungsten carbide has been analyzed using laser-pulsed atom probe tomography (APT). The influence of experimental parameters, including laser pulse energy, pulse repetition rate, and specimen base temperature, on the acquired data were evaluated from different aspects, such as mass spectrum, chemical composition, noise-to-signal ratio, and multiple events. Within all the applied analysis conditions, only 1 MHz pulse repetition rate led to a strong detector saturation effect, resulting in a largely biased chemical composition. A comparative study of the laser energy settings showed that an ~12 times higher energy was required for the less focused green laser of the LEAPTM 3000X HR system to achieve a similar evaporation field as the finer spot ultraviolet laser of the LEAPTM 5000 XS system.

Although acknowledged to be variable and subjective, manual annotation of cryo-electron tomography data is commonly used to answer structural questions and to create a “ground truth” for evaluation of automated segmentation algorithms. Validation of such annotation is lacking, but is critical for understanding the reproducibility of manual annotations. Here, we used voxel-based similarity scores for a variety of specimens, ranging in complexity and segmented by several annotators, to quantify the variation among their annotations. In addition, we have identified procedures for merging annotations to reduce variability, thereby increasing the reliability of manual annotation. Based on our analyses, we find that it is necessary to combine multiple manual annotations to increase the confidence level for answering structural questions. We also make recommendations to guide algorithm development for automated annotation of features of interest.

The article compares the relative stability of MCM-41 and related mesoporous materials in electron beam at an accelerating voltage of 100–300 kV. The work encountered in electron microscopy presents a comparison with similar research that has been carried out on nonporous and microporous silicates, especially α-quartz and zeolite Y. The trends in stability are analyzed, classifying the effects of sample preparation, organic and inorganic moieties, and electron accelerating voltage on beam stability. A higher synthesis temperature, the use of an acid catalyst in the synthesis, and the presence of additional organic or inorganic material within the channels were all found to stabilize these materials. The dose required to completely disrupt the structure increased with accelerating voltage for nearly all samples, suggesting a primarily radiolytic damage mechanism. The exception, MCM-41 containing nanometer-sized titania particles in its channels, was found to be almost insensitive to accelerating voltage.

Focused ion beam nanotomography (FIB-nt) is a novel method for high resolution three-dimensional (3D) imaging. In this investigation we assess the methodological parameters related to image acquisition and data processing that are critical for obtaining reproducible microstructural results from granular materials and from complex suspensions. For this purpose three case studies are performed: (1) The precision of FIB-nt is evaluated by analyzing a reference sample with nanospheres. Due to the implementation of an automated correction procedure, drift phenomena can be removed largely from the FIB data. However, at high magnifications remaining drift components can induce problems for 3D-shape reconstructions. (2) Correct object recognition from densely packed microstructures requires specific algorithms for splitting of agglomerated particles. To establish quantitative criteria for the correct degree of splitting, a parametric study with dry portland cement is performed. It is shown that splitting with a k-value of 0.6 leads to accurate results. (3) Finally, the reproducibility of the entire cryo-FIB analysis is investigated for high pressure frozen cement suspensions. Reproducible analyses can be obtained if the magnification is adapted to the particle size. At low magnifications the small particles and their surface area are underestimated. At high magnifications representativity is questioned because local inhomogeneities can become dominant.

In this work, we report that polarization second harmonic generation (PSHG) microscopy, commonly used in biomedical imaging, can quantitatively discriminate naturally aged from fresh starch-based glues used for conservation or restoration of paintings, works of art on paper, and books. Several samples of fresh and aged (7 years) flour and starch pastes were investigated by use of PSHG. In these types of adhesives, widely used in cultural heritage conservation, second harmonic generation (SHG) contrast originates primarily from the starch granules. It was found that in aged glues, the starch SHG effective orientation (SHG angle, θ) shifts to significantly higher values in comparison to the fresh granules. This shift is attributed to the different degree of granule hydration between fresh and aged adhesives. Thus noninvasive high-resolution nonlinear scattering can be employed to detect and quantify the degree of deterioration of restoration adhesives and to provide guidance toward future conservation treatments.

Nonwoven materials are found in a gamut of critical applications. This is partly due to the fact that these structures can be produced at high speed and engineered to deliver unique functionality at low cost. The behavior of these materials is highly dependent on alignment of fibers within the structure. The ability to characterize and also to control the structure is important, but very challenging due to the complex nature of the structures. Thus, to date, focus has been placed mainly on two-dimensional analysis techniques for describing the behavior of nonwovens. This article demonstrates the utility of three-dimensional (3D) digital volumetric imaging technique for visualizing and characterizing a complex 3D class of nonwoven structures produced by hydroentanglement.