Standardless EDS analysis of bulk and thin specimens

A direct relationship between the x-ray intensity ratio and the concentration ratio for bulk and thin specimens has been established by use of a revised full-diffusion model of electron scattering. The suitable ionization cross section, the most important parameter influencing the accuracy of the calculated Cliff-Lorimer factors, has been found after comparing the experimental intensity ratio I(L)/I(K) of eight elements (from Ge to Sn) and I(M)/I(L) of six elements (from Sm to Bi) with the calculated values. The quantitative standardless EDS analysis of bulk samples obtained by this direct method is more satisfactory than the commercial indirect method which gives the composition through ZAF correction after calculating the intensity factors of pure elements. The quantitative standardless analysis of thin samples has been improved by the suitable cross section significantly. This method has been applied to the analysis of film on substrate either without any common element or with one common element (P-Si glass film on Si). It has also been used to calculate the intensity factors of pure bulk samples and the backscattering correction factor in Auger electron spectroscopy.     

The microstructure of experimental SiC fibre-reinforced yttrium magnesium aluminosilicate (SiCf-YMAS) materials

Two experimental SiC fibre-reinforced yttrium magnesium aluminosilicate (SiCf-YMAS)-type ceramic-matrix composite (CMC) materials fabricated (i) by the glass process and (ii) by chemical precursor infiltration have been studied by light microscopy, transmission electron microscopy (TEM), high-resolution electron microscopy (HREM) and energy-dispersive X-ray spectroscopy (EDS). The distribution of the fibres inside the composite as well as the average diameter of fibres have been determined by image analysis. The microstructure of the YMAS matrices has been characterized by TEM observations. YMAS matrices are formed of two main phases, cordierite and β-yttrium silicate (Y₂Si₂O₇). Two minor phases (mullite and spinel) have been found to crystallize inside the cordierite and the yttrium silicate crystals. Fibre-matrix interfaces have been observed in HREM. A thin turbostratic carbon layer (20–30 nm) has been imaged in both composites at the fibre-matrix interface. It crystallizes along the matrix interface and grows inside the fibre, forming a diffuse interphase. The carbon layer is believed to be the consequence of reaction between oxygen in the matrix and SiC nanocrystals of the Nicalon fibres.     

Microdiffraction,stem imaging and ELS at crystal surfaces

The interaction of electrons with the flat face of a small crystal, when the beam is almost parallel to the face, is strongly influenced by the form of the potential field at the surface and the excitations associated with the surface. Microdiffraction patterns show evidence of refraction displacements from which information on the form of the potential distribution can be deduced. Channeling of the electrons along the surface gives additional spots in the microdiffraction patterns and fringes of varying periodicity in the image. The generation of transition radiation as the electron is accelerated in the vicinity of the surface gives rise to prominent characteristic features in the 0–30 eV range of the energy loss spectra. Also the peaks associated with excitations of surface electronic states are strong.     

TEM/STEM requirements: multipurpose or specialized instruments?

The relative advantages and disadvantages of high resolution and other more general imaging techniques are considered and the potential of a multipurpose, moderate resolution, TEM with EELS is assessed.     

HREM and STEM of defects in multiply-twinned particles

The structure of defects in multiply-twinned particles has been studied in detail using high-resolution lattice imaging, dark field and microdiffraction techniques. Icosahedral particles with sizes greater than about 15 nm were found to contain defects, in the form of stacking fault loops parallel with the external surface, which were extremely difficult to detect by conventional amplitude contrast techniques. Microdiffraction mappings correlated with these results, showing large rotations of the face-centred cubic segments. Results for decahedral particles indicated the presence of stacking faults running adjacent to, and parallel with, the twin boundaries. Microdiffraction maps confirmed that the particle structure was face-centred cubic, and also verified that the apparent epitaxy of these particles was highly variable. Models for the defects are proposed and discussed, and the relative merits of HREM and STEM for elucidating structural details in small particles is briefly considered. Finally, the potential for direct imaging at surfaces, as demonstrated by some recent images, is discussed.     

HREM and STEM of intergranular films at zinc oxide varistor grain boundaries

Grain boundaries in model ZnO–Bi₂O₃ and ZnO–Bi₂O₃–CoO varistors and a commercial multicomponent varistor have been characterized by high-resolution electron microscopy (HREM) and scanning transmission electron microscopy (STEM), in order to determine the relationship between Bi grain boundary segregation and formation of thin intergranular films. By controlling Bi₂O₃ content, applied pressure and temperature, the grain boundary Bi excess has been systematically varied from nearly zero to Γ_Bi = 1 × 10¹⁵ cm⁻² (≈ 1 monolayer), as measured by HB 603 STEM using an area-scan method. HREM shows that intergranular amorphous films are clearly distinguishable in samples with Γ_Bi > 8 × 10¹⁴ cm⁻². These films range in thickness, depending on the Bi excess, from 0.6 to 1.5 nm. Similar films of ≈ 1 nm thickness are widely observed in the commercial varistor. The composition of the films is a ZnO–Bi₂O₃ solid solution, which is in all cases more enriched in ZnO than the bulk eutectic liquid. The Bi-doped grain boundaries in ZnO varistors therefore contain an intergranular amorphous film which has not only an equilibrium thickness, but also a distinct equilibrium composition.     

Persistent misconceptions about incoherence in electron microscopy

Incoherence in electron microscopic imaging occurs when during the observation the microscope and the object are subject to fluctuations. In order to speed up the computer simulation of the images, approximations are used that are considered as valid. In this paper we will question the validity of these approximations and show that in specific cases they can lead to erroneous results.It is shown in particular in the case of one single vibrating atom that the thermal diffuse scattering that causes the signal in HAADF STEM is not only dependent on Z but also on the mean square displacement of the atom so that it can even be large for light atoms in soft matter, provided the right HAADF aperture is used.In HREM imaging the diffuse scattering leaks out of the coherent (elastic) wave and is redistributed in the background. This might explain the mismatch in elastic contrast (Stobbs factor) especially for crystals with a thickness beyond the extinction distance, where also the HAADF signal saturates and the elastic (coherent) component vanishes.     

Preparing and analyzing fractured archaeological fibers

A technique was developed to prepare archaeological fiber cross sections for electron microscopic examination and x-ray analysis. Use of this new method allows chemical and morphological information to be obtained from the interior of a single fiber or yarn. Fibers are fractured while frozen and then freeze dried. Following mounting and carbon coating, fibers are examined by scanning and backscatter electron microscopy and then analyzed by using energydispersive spectrometry. Elemental distribution is mapped by using image-processing software. In this report, the described technique is employed in the examination of ancient fibers from three different long-term storage environments (moist buried, dry buried, museum stored). Data obtained by examining the interior of fibers such as these provide insight into the conditions of a fiber's growth, the treatments applied during the fiber's processing and use, and the conditions in which the fiber was stored.     

High-resolution rlectron microscopic investigation of supported platinum particles reduced at high temperatures

High-resolution electron microscopy has been used to characterize the platinum particles supported on TiO₂ or ZnO. After reduction at elevated temperatures, the metallic particles display a regular, faceted shape, and several superstructures, Pt₃ Ti(C), Pt₃ Ti(H), PtTi, and PtZn, have been found. These results, which may involve strong metal-support interaction, have been confirmed by optical diffraction and image simulation.     

Freeze-fracturing at low temperatures: I. A device for fracturing biological specimens at 77–10 K under high vacuum

Standard freeze-etching or freeze-cleaving is performed at 173 K in a vacuum of 133 μPa or at 77 K under liquid nitrogen with subsequent transfer of the specimen into a vacuum chamber. It has been suggested that the frequent poor resolution of morphological details, the poor complementarity of innermembrane protein particles and the semi-crystalline substructures in biomembranes are caused by structural distortion or plastic deformation due to sheer forces which occur even at 77 K during fracturing or cleaving. In addition, water contamination and radiant heat damage occurring during replication introduce artefacts to the structural record. These artefacts could be avoided or reduced by lowering the temperature at which fracturing or cleaving and shadowing is carried out, to about 10 K. Therefore, a device for cleaving biological specimens at 15–10 K under high vacuum was constructed. To allow the use of existing equipment, the device was built into a standard Balzers 301 vacuum unit, where the specimen transfer is done via an airlock system which allows hoar frost contamination free transport of the specimen holder onto the specimen table. To reduce or prevent the condensation of water and other residual gases in the vacuum onto the freshly cleaved specimen surface at 10 K, the specimen is surrounded by two cooled surfaces of 6 and 20 K. All condensable gases outside those shielding shrouds will condense on these surfaces before reaching the specimen. This makes it possible to work at a high vacuum of 3 μPa outside the cooled shrouds, which can be reached with standard turbomolecular pumps. The actual vacuum within the cooled shrouds is estimated to be approximately 13 nPa. Residual gas analysis before and during replication reveals equal conditions to ultra high vacuum systems. An analysis of the yeast cell paracrystalline plasmalemma structure shows that the topographic resolution of the crystalline arrays has been improved by working at 12 K. However, plastic deformation still occurs under these conditions. This observation points to the possibility that what is described as plastic deformation, for at least some membrane proteins, may be a loss of resilience at low temperatures.     

High-resolution electron microscopy of human enamel crystals

The structure of enamel crystals obtained from four human premolars has been studied by high-resolution electron microscopy (HREM) in the [0001], [***2110], [***1540], [***0110] and [***1213] crystallographic directions at various microscope defocus and crystal thickness values. The resolution obtained has not previously been reported for human enamel crystals. In all cases, it was possible to match the experimental images to images calculated using the atomic positions of mineral hydroxyapatite. However, a deviation from hexagonal symmetry characterized by marked (***1010) planes of intensity different from the one of the (***3030) and {***1010}-type planes was observed. In this work, we present an improvement of Scherzer resolution of 0.25-0.20 nm over previous work on biological enamel crystals. This improvement of resolution has permitted the incorporation of crystallographic reflections of higher spatial frequencies into the imaging process of the microscope and has led to a more precise structure determination of the crystals studied.     

Procedures for focusing,stigmating and alignment in high resolution electron microscopy

Several different approaches to the alignment, stigmating and focusing of a high resolution electron microscope are evaluated theoretically and experimentally. Ambiguities in the interpretation of diffractograms are pointed out which make it necessary to explore a range of incident beam directions before the correct alignment can be established. The variation of image contrast with the imaging conditions is examined in detail, and its global minimization is shown to be a reliable criteria for adjustment of all parameters. Recommendations are made as to the best procedures to adopt in various circumstances, and a computer-controlled procedure based on the image contrast is described which sets all parameters automatically in less than a minute.     

On high-resolution transmission electron microscopy in an unstable magnetic environment

Owing to the particular environment of the JEM4000EX HREM at the University of Pennsylvania, it became necessary to control the magnetic environment around the microscope. The unstable magnetic environment included two components, a slowly wandering vertical DC magnetic field and an AC magnetic field. The effects of these fields on the microscope performance were to introduce an uncertainty in the objective lens defocus value over a series of images and to reduce the attainable resolution of the microscope, respectively. A solution is presented in which these fields are stably reduced well within the limits of sensitivity of the JEOL, Ltd. JEM4000EX for high-resolution imaging. This solution is based on a feedback loop using a pseudo-Helmholtz coil to generate a well-defined vertical magnetic field.     

A review of on-line processing of chemical analysis data using a conventional CTEM/STEM

A review of the on-line processing of X-rays and the energetically complementary electron intensities to yield chemical compositions is followed by a discussion of X-ray mapping, energy filtered images and some extended uses of conventionally supplied equipment associated with these analytic techniques.     

X-ray microanalysis of freeze-dried and frozen-hydrated cryosections

The elemental composition and the ultrastructure of biological cells were studied by scanning transmission electron microscopy (STEM) combined with energy dispersive X-ray microanalysis. The preparation technique involves cryofixation, cryoultramicrotomy, cryotransfer, and freeze-drying of samples. Freeze-dried cryosections 100-nm thick appeared to be appropriate for measuring the distribution of diffusible elements and water in different compartments of the cells. The lateral analytical resolution was less than 50 nm, depending on ice crystal damage and section thickness. The detection limit was in the range of 10 mmol/kg dry weight for all elements with an atomic number higher than 12; for sodium and magnesium the detection limits were about 30 and 20 mmol/kg dry weight, respectively. The darkfield intensity in STEM is linearly related to the mass thickness. Thus, it becomes possible to measure the water content in intracellular compartments by using the darkfield signal of the dry mass remaining after freeze-drying. By combining the X-ray microanalytical data expressed as dry weight concentrations with the measurements of the water content, physiologically more meaningful wet weight concentrations of elements were determined. In comparison to freeze-dried cryosections frozen-hydrated sections showed poor contrast and were very sensitive against radiation damage, resulting in mass loss. The high electron exposure required for recording X-ray spectra made reproducible microanalysis of ultrathin (about 100-nm thick) frozen-hydrated sections impossible. The mass loss could be reduced by carbon coating; however, the improvement achieved thus far is still insufficient for applications in X-ray microanalysis. Therefore, at present only bulk specimens or at least 1-μm thick sections can be used for X-ray microanalysis of frozen-hydrated biological samples.     

Optimized conditions for imaging the effects of bonding charge density in electron microscopy

We report on the observability of valence bonding effects in aberration-corrected high resolution electron microscopy (HREM) images along the [0 1 0] projection of the mineral Forsterite (Mg₂SiO₄). We have also performed exit wave restorations using simulated noisy images and have determined that both the intensities of individual images and the modulus of the restored complex exit wave are most sensitive to bonding effects at a level of 25% for moderately thick samples of 20-25 nm. This relatively large thickness is due to dynamical amplification of bonding contrast arising from partial de-channeling of 1s states. Simulations also suggest that bonding contrast is similarly high for an un-corrected conventional electron microscope, implying an experimental limitation of signal to noise ratio rather than spatial resolution.     

Microstructure of Sm5Ni19 intermetallic compound observed by high-resolution electron microscope

The method of high-resolution electron microscopy is applied to study the microstructure of the Sm–Ni intermetallic compound whose composition is near the Sm₅Ni₁₉. Several new polytypic structures (5T, 7T, 15R and 18R) are found and their crystal structures are analysed by high-resolution imaging method combined with electron diffraction method. Various kinds of defects are observed and the atomic arrangements near the defects are deduced; the defects include stacking faults in Sm₅Ni₁₉, intergrowth of the Sm₂Ni₇ or other complex structures in between the matrix of Sm₅Ni₁₉, complicated defect structure which changes the stacking sequence of Sm₅Ni₁₉, and lattice defects in the region of terminating the intergrowth.     

Chemical and morphological changes in human dentin after Er:YAGlaser irradiation: EDS and SEM analysis

Sixty samples of human dentin were divided into six groups (n = 10) and were irradiated with Er:YAG laser at 100 mJ–19.9 J/cm², 150 mJ–29.8 J/cm², 100 mJ–35.3 J/cm², 150 mJ–53.0 J/cm², 200 mJ–70.7 J/cm², and 250 mJ–88.5 J/cm², respectively, at 7 Hz under a water spray. The atomic percentages of carbon, oxygen, magnesium, calcium, and phosphorus and the Ca‐to‐P molar ratio on the dentin were determined by energy dispersive X‐ray spectroscopy. The morphological changes were observed using scanning electron microscopy. A paired t‐test was used in statistical analysis before and after irradiation, and a one‐way ANOVA was performed (P ≤ 0.05). The atomic percent of C tended to decrease in all of the groups after irradiation with statistically significant differences, O and Mg increased with significant differences in all of the groups, and the Ca‐to‐P molar ratio increased in groups IV, V, and VI, with statistically significant differences between groups II and VI. All the irradiated samples showed morphological changes. Major changes in the chemical composition of dentin were observed in trace elements. A significant increase in the Ca‐to‐P ratio was observed in the higher energy density groups. Morphological changes included loss of smear layer with exposed dentinal tubules. The changes produced by the different energy densities employed could have clinical implications, additional studies are required to clarify them. Microsc. Res. Tech. 78:1019–1025, 2015. © 2015 Wiley Periodicals, Inc.     

High-resolution electron microscopy and its applications

A review of research on high-resolution electron microscopy (HREM) carried out at the Institute of Physics, the Chinese Academy of Sciences, is presented. Apart from the direct observation of crystal and quasicrystal defects for some alloys, oxides, minerals, etc., and the structure determination for some minute crystals, an approximate image-contrast theory named pseudo-weak-phase object approximation (PWPOA), which shows the image contrast change with crystal thickness, is described. Within the framework of PWPOA, the image contrast of lithium ions in the crystal of R-Li₂Ti₃O₇ has been observed. The usefulness of diffraction analysis techniques such as the direct method and Patterson method in HREM is discussed. Image deconvolution and resolution enhancement for weak-phase objects by use of the direct method are illustrated. In addition, preliminary results of image restoration for thick crystals are given.     

Structure of superconducting thin films of YBa2Cu3O7−x grown on SrTiO3 and cubic zirconia

Thin films of the superconductive oxide YBa₂Cu₃O_7?x have been made by electron-beam coevaporation of the metals in an oxygen atmosphere onto single-crystal {001}-oriented SrTiO₃ and yttria-stabilized zirconia (YSZ) substrates. The oxide films were superconducting in the as-deposited state (T_c = 81–83K, J_c = 10⁶ A/cm² at 4.2K). Bright-field imaging, selectedarea diffraction (SAD), and high-resolution imaging in the transmission electron microscope were used to characterize the microstructure of these films. All of the films were polycrystalline. On SrTiO₃ the films were oriented, for the most part, with {110} parallel to the substrate surface. On YSZ, two microstructures were observed: one with smaller rectangular grains oriented with (100) or (010) parallel to the substrate surface and the other with (001) parallel to the surface (i.e., c-axis up).