While electron cryotomography (ECT) provides “molecular” resolution, three-dimensional images of unique biological specimens, sample crowdedness, and/or resolution limitations can make it difficult to identify specific macromolecular components. Here we used a 1.4 nm Nanogold® cluster specifically attached to the Fc fragment of IgG to monitor its interaction with the neonatal Fc receptor (FcRn), a membrane-bound receptor that transports IgG across cells in acidic intracellular vesicles. ECT was used to image complexes formed by Nanogold-labeled Fc bound to FcRn attached to the outer surface of synthetic liposomes. In the resulting three-dimensional reconstructions, 1.4 nm Nanogold particles were distributed predominantly along the interfaces where 2:1 FcRn-Fc complexes bridged adjacent lipid bilayers. These results demonstrate that the 1.4 nm Nanogold cluster is visible in tomograms of typically thick samples (∼250 nm) recorded with defocuses appropriate for large macromolecules and is thus an effective marker.

Microscopic images of hair from 36 individuals were evaluated for color (using the CIE L*a*b* color space), hair thickness, and the pattern of pigmentation. These measurements were compared to each other and to reflective spectrophotometric measurements of macroscopic hair color. Three areas on three strands of hair were examined for each individual. Despite a large ΔE* value, there is a strong relationship between macroscopic and microscopic L* values that would be useful if one was trying to predict the color of a head of hair (macroscopic) from a strand (microscopic). A relationship between hair lightness (L*) and thickness is also found at both the macroscopic and microscopic levels. The information acquired in this study may be useful to those conducting a microscopic examination of hair, especially in forensic science, as well as those researchers investigating macroscopic hair color and hair morphology for the purposes of forensic science, anthropology, and medical science. Further investigation into objectively measuring and quantifying the microscopic color and properties of hair will assist in increasing its validity as evidence in forensic casework.

With light and scanning electron microscopy of vascular corrosion casts, we have observed in Urobatis jamaicensis that the fourth epibranchial arteries do not merge completely with the dorsal aorta. Instead they form a brief anastomosis with a short vessel projecting ventrally from the dorsal aorta and maintain their integrity as separately distinct vessels. Posterior to the anastomosis, the right epibranchial becomes the celiac trunk and left epibranchial becomes the anterior mesenteric artery/posterior intestinal artery. This vascular configuration appears to be unique in elasmobranchs.

Electron backscatter diffraction (EBSD) is becoming a widely used technique to determine crystallographic orientation in biogenic carbonates. Despite this use, there is little information available on preparation for the analysis of biogenic carbonates. EBSD data are compared for biogenic aragonite and calcite in the common blue mussel, Mytilus edulis, using different types of resin and thicknesses of carbon coating. Results indicate that carbonate biomineral samples provide better EBSD results if they are embedded in resin, particularly epoxy resin. A uniform layer of carbon of 2.5 nm thickness provides sufficient conductivity for EBSD analyses of such insulators to avoid charging without masking the diffracted signal. Diffraction intensity decreases with carbon coating thickness of 5 nm or more. This study demonstrates the importance of optimizing sample preparation for EBSD analyses of insulators such as carbonate biominerals.

An in situ electron energy loss spectroscopy (EELS) technique has been developed to investigate the dynamic processes associated with electron-beam nanofabrication on thin membranes. In this article, practical applications germane to e-beam nanofabrication are illustrated with a case study of the drilling of nanometer-sized pores in silicon nitride membranes. This technique involves successive acquisitions of the plasmon-loss and the core-level ionization-loss spectra in real time, both of which provide the information regarding the hole-drilling kinetics, including two respective rates for total mass loss, individual nitrogen and silicon element depletion, and the change of the atomic bonding environment. In addition, the in situ EELS also provides an alternative method for endpoint detection with a potentially higher time resolution than by imaging. On the basis of the time evolution of in situ EELS spectra, a qualitative working model combining knock-on sputtering, irradiation-induced mass transport, and phase separation can be proposed.

This article presents a (scanning) transmission electron microscopy (TEM) study of Mn valency and its structural origin at La0.7Sr0.3MnO3/SrTiO3(0 0 1) thin film interfaces. Mn valency deviations can lead to a breakdown of ferromagnetic order and thus lower the tunneling magnetoresistance of tunnel junctions. Here, at the interface, a Mn valency reduction of 0.16 ± 0.10 compared to the film interior and an additional feature at the low energy-loss flank of the Mn-L3 line have been observed. The latter may be attributed to an elongation of the (0 0 1) plane spacing at the interface detected by geometrical phase analysis of high-resolution images. Regarding the interface geometry, high-resolution high-angle annular dark-field scanning TEM images reveal an atomically sharp interface in some regions whereas the transition appears broadened in others. This can be explained by the presence of steps. The performed measurements indicate that, among the various structure-related influences on the valency, the atomic layer termination and the local oxygen content are most important.

A method for the analysis of cathodoluminescence spectra is described that enables quantitative trace-element-level distributions to be mapped within minerals and materials. Cathodoluminescence intensities for a number of rare earth elements are determined by Gaussian peak fitting, and these intensities show positive correlation with independently measured concentrations down to parts per million levels. The ability to quantify cathodoluminescence spectra provides a powerful tool to determine both trace element abundances and charge state, while major elemental levels can be determined using more traditional X-ray spectrometry. To illustrate the approach, a scheelite from Kalgoorlie, Western Australia, is hyperspectrally mapped and the cathodoluminescence is calibrated against microanalyses collected using a laser ablation inductively coupled plasma mass spectrometer. Trace element maps show micron scale zoning for the rare earth elements Sm3+, Dy3+, Er3+, and Eu3+/Eu2+. The distribution of Eu2+/Eu3+ suggests that both valences of Eu have been preserved in the scheelite since its crystallization 1.63 billion years ago.

A detector that looks promising for advanced imaging modalities—such as X-ray absorption contrast imaging, X-ray fluorescence imaging, and diffraction-enhanced imaging—is the controlled-drift detector (CDD). The CDD is a novel two-dimensional X-ray imager with energy resolving capability of spectroscopic quality. It is built on a fully depleted silicon wafer and features fast readout while being operated at or near room temperature. The use of CDDs in the aforementioned applications allows translating these techniques from synchrotron-based experiments to laboratory-size experiments using polychromatic X-ray generators. We have built a dedicated and versatile detection module based on a 36 mm2 CDD chip featuring pixels of 180 × 180 μm2, and we evaluated the system performance in different X-ray imaging applications both with synchrotron-based experiments and in the laboratory environment.

The International Technology Roadmap for Semiconductors ranks dopant profiling as one of the most difficult challenges for analysis of semiconductors. Dopant mapping in the scanning electron microscope (SEM) has the potential to provide a solution. This technique has not yet found widespread application, however, mainly due to the lack of a comprehensive theoretical model, uncertain quantification, and its inability to differentiate doping levels in n-type silicon. Although a Monte Carlo model was recently published that closely matched experimental data obtained in p-doped silicon to data obtained from the theoretical model, a large discrepancy between experimental data obtained for n-type silicon was found. Here we present a Monte Carlo model that provides close matches between experimental and calculated data in both n- and p-type silicon, paving the way for a widespread application of SEM dopant contrast.

Electrical interconnects in integrated circuits have shrunk to sizes in the range of 20–100 nm. Accurate measurements of the dimensions of these nanowires are essential for identifying the dominant electron scattering mechanisms affecting wire resistivity as they continue to shrink. We report a systematic study of the effect of line edge roughness on the apparent cross-sectional area of 90 nm Cu wires with a TaN/Ta barrier measured by conventional two-dimensional projection imaging and three-dimensional electron tomography. Discrepancies in area measurements due to the overlap of defects along the wire's length lead to a 5% difference in the resistivities predicted by the two methods. Tomography of thick cross sections is shown to give a more accurate representation of the original structure and allows more efficient sampling of the wire's cross-sectional area. The effect of roughness on measurements from projection images is minimized for cross-section thicknesses less than 50 nm, or approximately half the spatial frequency of the roughness variations along the length of the investigated wires.

Two purely carbon-based functional polymer systems were investigated by bright-field conventional transmission electron microscopy (CTEM) and high-angle annular dark-field scanning transmission electron microscopy (HAADF-STEM). For a carbon black (CB) filled polymer system, HAADF-STEM provides high contrast between the CB agglomerates and the polymer matrix so that details of the interface organization easily can be revealed and assignment of the CB phase is straightforward. For a second system, the functional polymer blend representing the photoactive layer of a polymer solar cell, details of its nanoscale organization could be observed that were not accessible with CTEM. By varying the camera length in HAADF-STEM imaging, the contrast can be enhanced between crystalline and amorphous compounds due to diffraction contrast so that nanoscale interconnections between domains are identified. In general, due to its incoherent imaging characteristics HAADF-STEM allows for reliable interpretation of the data obtained.

A numerical and experimental analysis of the optical beam deflection system used to monitor microcantilevers subjected to simultaneous deflection and twisting such as in lateral or frictional force microscopy was performed. This study focused on two optical beam deflection orientations where in the first case the optical beam and the detector are at a right angle to the length of the cantilever and the second case, which is the more standard orientation, the optical beam is parallel to the length of the lever. This study finds that it is possible to model the twist and the deflection separately and treat each motion independently. Simulations have shown that the above-mentioned systems are equivalent in accuracy and sensitivity for monitoring the simultaneous twist and deflection of cantilevers.