Reflection electron microscopy methods for the study of surface structure

The techniques of reflection electron microscopy (REM) using TEM instruments and scanning reflection electron microscopy (SREM) using STEM instruments have been explored as means for the observation of surface structure with high spatial resolution, better than 1 nm in each case. Under the ordinary environment of a commercial TEM instrument, we have studied the contrast in REM images of atomic steps and made comparison with the calculated results from the multi-slice dynamical diffraction theory. Comparison has also been made between the REM images of defects and the calculated images based on the column approximation. The influence of surface resonances on the contrast has been investigated. By SREM performed in a modified HB5 STEM with attached high vacuum preparation chamber, we have observed the formation of periodically distributed Pd particles on the surface of cleaved MgO.     

Identifying threading dislocations in GaN films and substrates by electron channelling

Electron channelling contrast imaging of threading dislocations in GaN (0002) substrates and epitaxial films has been demonstrated using a conventional polepiece-mounted backscatter detector in a commercial scanning electron microscope. The influence of accelerating voltage and diffraction vector on contrast features denoting specific threading dislocation types has been studied. As confirmed by coordinated transmission electron microscopy analysis, electron channelling contrast imaging contrast features for edge-type threading dislocations are spatially smaller than mixed-type threading dislocations in GaN. This ability to delineate GaN edge threading dislocations from mixed type was also confirmed by defect-selective etch processing using molten MgO/KOH. This study validates electron channelling contrast imaging as a nondestructive and widely accessible method for spatially mapping and identifying dislocations in GaN with wider applicability for other single-crystal materials.     

Measurements of plastic strain in copper due to sliding wear

Wear experiments have been conducted to determine the plastic strains that are introduced in surface material near sliding wear tracks. Both oillubricated and dry-sliding experiments have been carried out at different sliding distances on surfaces of copper. The strain values were determined from selected-area electron channelling patterns obtained from regions as small as 10 μm in size and 0.05 μm deep around the wear track using a scanning electron microscope. A deformed calibration specimen was used to relate the electron channelling band contrast to the deformation strain. Strain maps were obtained on the wear surface lateral to the wear track and also below the surface using electropolishing metal removal techniques. Particular attention was given to the near-surface strain values. In all cases the maximum strain was found at the wear surface located at the track center and the strains decreased uniformly with depth. Significant large strains were also found outside the wear tracks. The results are compared with those previously reported for iron and with other relevant studies.     

Fine structure of the images of atoms in 011-gold crystals in various Bragg reflecting conditions

Fine structures with petal and horseshoe shape contrast have been observed in the high resolution electron microscope images of atoms in a 011 gold crystal in deviated Bragg reflecting conditions. The images with such fine structures have been recorded by using only the beams of 000, four 111 and two 200 reflections with axial illumination. The contrast of the images has been discussed by the many beam dynamical theory of electron diffraction and the image formation theory considering the spatial and temporal coherence of the electron waves. It has been noted that the temporal coherence has much more effect on the change of image contrast than the spatial coherence. The imaging conditions have been analysed in terms of Fourier images and the aberration free focus conditions.     

Design of a microfabricated, two-electrode phase-contrast element suitable for electron microscopy

A miniature electrostatic element has been designed to selectively apply a 90 degrees phase shift to the unscattered beam in the back focal plane of the objective lens, in order to realize Zernike-type, in-focus phase contrast in an electron microscope. The design involves a cylindrically shaped, biased-voltage electrode, which is surrounded by a concentric grounded electrode. Electrostatic calculations have been used to determine that the fringing fields in the region of the scattered electron beams will cause a negligible phase shift as long as the ratio of electrode length to the transverse feature size is greater than 5:1. Unlike the planar, three-electrode einzel lens originally proposed by Boersch for the same purpose, this new design does not require insulating layers to separate the biased and grounded electrodes, and it can thus be produced by a very simple microfabrication process. Scanning electron microscope images confirm that mechanically robust devices with feature sizes of approximately 1 microm can be easily fabricated. Preliminary experimental images demonstrate that these devices do apply a 90 degrees phase shift between the scattered and unscattered electrons, as expected.     

Backscattered electron imaging and scanning transmission electron microscopy imaging of multi-layers

Experimental and theoretical results on image contrast of semiconductor multi-layers in scanning electron microscopy investigation are reported. Two imaging modes have been considered: backscattered electron imaging of bulk specimen and scanning transmission imaging of thinned specimens. The following main results have been reached. The image resolution of the multi-layers is, in both cases, defined by the probe size. The contrast, governed by density and atomic number differences, is affected by the size of the interaction volume in backscattered electron imaging and by the beam broadening in scanning transmission. Operating in the scanning transmission mode, the contrast of bright field images can be easily related to local variation in atomic number and density of the specimen while the dark field image contrast is strongly affected by electron beam energy, detector collection angles and specimen thickness. All these factors are able to produce contrast reversals that are difficult to explain without the support of a suitable simulation code.     

The contribution of phonon scattering to high-resolution images measured by off-axis electron holography

The contribution of electrons that have been phonon scattered to the lattice fringe amplitude and the background intensity of a high-resolution electron microscope (HREM) image is addressed experimentally through the analysis of a defocus series of energy-filtered off-axis electron holograms. It is shown that at a typical specimen thickness used for HREM imaging approximately 15% of the electrons that contribute to an energy-filtered image have been phonon scattered. At this specimen thickness, the phonon-scattered electrons contribute a lattice image of opposite contrast to the elastic lattice image. The overall lattice fringe contrast is then reduced to 70% of the value that it would have in the absence of phonon scattering. At higher specimen thickness, the behaviour is defocus-dependent, with the phonon image having lattice fringe contrast of either the same or the opposite sense to the elastic image as the defocus is varied.     

Quantitative phase-amplitude microscopy II: differential interference contrast imaging for biological TEM

Although phase contrast microscopy is widespread in optical microscopy, it has not been as widely adopted in transmission electron microscopy (TEM), which has therefore to a large extent relied on staining techniques to yield sufficient contrast. Those methods of phase contrast that are used in biological electron microscopy have been limited by factors such as the need for small phase shifts in very thin samples, the requirement for difficult experimental conditions, or the use of complex data analysis methods. We here demonstrate a simple method for quantitative TEM phase microscopy that is suitable for large phase shifts and requires only two images. We present a TEM phase image of unstained Radula sp. (liverwort spore). We show how the image may be transformed into the differential interference contrast image format familiar from optical microscopy. The phase images contain features not visible with the other imaging modalities. The resulting technique should permit phase contrast TEM to be performed almost as readily as phase contrast optical microscopy.     

Contrast In Transmission Electron Micrographs Of Void Arrays In Crystals

In order to understand the detailed features in electron micrographs from radiation induced void lattices in crystals, an attempt has been made to determine the contrast from columns of regularly spaced strain-free spherical voids using calculations based on the two-beam approximation. Analytical expressions have been obtained for the wave amplitudes at the exit surface of a model crystal using matrix algebra. Line profiles for the contrast across a void column have been computed and two-dimensional images simulated on a line printer. The contrast behaviour has been analysed in terms of deviation from the Bragg condition for varying void column parameter: void distance/void radius. For dynamical, as well as for kinematical diffraction conditions the influence on the image of defocusing phase contrast has been investigated by addition of extra phase contributions to the diffracted beam amplitudes. Defocusing in most cases induces an enhanced contrast from the void column.     

Morphology of organic electronic materials imaged via electron tomography

Organic electronic materials and nanostructures have been studied with the use of electron tomography. Nanostructured materials including contrast enhancing features have been studied and double tilt data collection has been employed to improve reconstructions. Tomography reconstructions of active layers of organic solar cells, where various preparation techniques have been used, have been analysed and compared to device behaviour. Small changes in preparation procedures may lead to large differences in morphology and device performance, and the results also indicate a complex relation between these.     

STEM imaging of monatomic surface steps and emergent dislocations

Surface features such as dislocations and monatomic steps have previously been imaged using reflection microscopy with conventional transmission electron microscopes. Here it is shown that similar features can be observed with a scanning transmission electron microscope by using phase contrast. Bragg contrast is very small unless a displaced aperture technique is employed.     

Aperture contrast in thick amorphous specimens using scanning transmission electron microscopy

The contrast observed in thick amorphous specimens using a scanning transmission electron microscope (STEM) can be considerably improved by the use of an optimum collector aperture angle. The size of this angle can be calculated by considering the variation of electron current transmitted through the specimen as a function both of the specimen thickness and of the angle of collection subtended at the specimen. Typically these calculations predict optimum angles to be several times the half-width of the elastic scattering distribution, often 10(-1) rad or more. Observations of biological sections of up to 2 micron in thickness using scanning attachments of commercial transmission microscopes have verifie these results at beam voltages of 50, 100 and 200 kV. Wide angle convergent beam diffraction patterns were used to give accurate values of the effective angles represented by the various collector apertures. Once the linearity of the detector-amplifier system had been established, operation in a line modulation mode enabled quantitative measurements to be made of the image contrast. Such measurements also offer a quick effective method of comparing electron beam penetrations.     

Assessment of electron irradiation damage to biomolecules using the Patterson function

A method for the assessment of electron radiation damage to 5-iodouracil is described which involves the use of the Patterson function. Changes in Patterson maps computed from the electron diffraction patterns recorded at increasing electron irradiation have been related to structural and chemical damage to 5-iodouracil. The process of radiation damage is also discussed in terms of a disorder parameter which was found to increase with increasing irradiation.     

TEM characterization of self-organized CdSe/ZnSe quantum dots

CdSe quantum dots (QDs) grown on ZnSe were investigated by various transmission electron microscopy (TEM) techniques including diffraction contrast imaging, high-resolution and analytical transmission electron microscopy both of plan-view as well as cross-section specimens. The size of the QDs ranges from about 5–50 nm, where from the contrast features in plan-view imaging two classes can be differentiated. In the features of the smaller dots there is no inner fine structure resolvable. The larger ones exhibit contrast features of fourfold symmetry as expected for pyramid-like islands. Corresponding simulations of diffraction contrast images of truncated CdSe pyramids with the edges of the basal plane orientated parallel to <100> are in relatively good agreement with this assumption. In TEM diffraction contrast imaging of cross-section samples the locations of the quantum dots are visualized by additional dark contrast features. The QDs have a distinct larger extension in growth direction compared to the almost uniformly thick CdSe wetting layer. The presence of the CdSe QDs was also confirmed by energy-dispersive X-ray spectroscopy.     

Application of the scatter diagram technique to the scanning electron microscope: preliminary results from diamond

Using primary beam energies E0 ranging from 0.2 to 15 keV and an in-lens detector, a series of images of the same region of an artificial microstructured diamond sample have been acquired in scanning electron microscopy. Next, the images were analysed by using a scatter diagram technique to underline the topographic contrast change and contrast reversal. The results obtained from 0.5 to 15 keV are discussed with the help of an expression derived from the constant loss model for the secondary electron yield delta of diamond, but including the respective roles of the angle of incidence, i, and of the angle of detection, alpha. More surprising is the quality of images obtained at a beam energy as low as 0.2 keV, and more difficult to explain is the significant contrast change between 0.2 keV and 0.5 keV. For the first time, scatter diagrams are used as a diagnostic tool in scanning electron microscopy, and after some improvements it is hoped that the experimental approach followed here may lead to quantitative estimates of the local tilts of a specimen surface.     

Lattice fringe studies of (100) defects in enamel crystallites

Lattice imperfections in the structure of shark (Nataprion brevirostrais) enamel crystallites have been studied by means of lattice-imaging in a high-resolution transmission electron microscope. Line defects exhibiting various lengths (100–2500 Å) and widths (10–30 Å) and demonstrating phase behaviour different from that of the host crystal were observed along the fringes representing (100) lattice planes. When defect sites were subjected to an electron beam, an annealing process was observed in which the difference in phase contrast eventually disappeared and the defect itself became an intricate part of the (100)-type lattice planes. A through-focal series of the defects show a reversal in phase contrast. By examining the contrast behaviour under certain focal conditions and by adopting the Fresnel diffraction mode, the refractive index of the defect regions was determined to be lower than that of the apatite crystal. Present findings suggest that the interlayering structure along the (100) planes is responsible for the line defects observed in enamel apatites.     

Application of Nomarski interference contrast microscopy as a thickness monitor in the preparation of transparent,SiC-based,cross-sectional TEM samples

Reflected light optical microscopy using a Nomarski prism and a differential interference contrast filter have been employed in concert to achieve a technique that provides an accurate color reference for thickness during the dimpling and ion milling of transparent transmission electron microscopy samples of 6H-SiC(000 1) wafers. The samples had thin films of AIN, GaN, and Au deposited on the SiC substrate. A sequence of variously colored primary and secondary interference bands was observed when the SiC was thinner than 20 microm using an optical microscope. The color bands were correlated with the TEM sample thickness as measured via scanning electron microscopy. The interference contrast was used to provide an indication of the dimpling rate, the ion milling rate, and also the most probable location of perforation, which are useful to reduce sample breakage. The application of pressure during the initial cross-sectional preparation reduced the separation of the two halves of the sample sandwich and resulted in increased shielding of the film surface from ion milling damage.     

Scanning electron microscopy studies of double-layer silicon structures prepar by epitaxy and direct bonding

The electrical properties of multilayer structures obtained by direct bonding of silicon wafers and epitaxial growth have been investigated. The measurements were made by scanning electron microscopy (SEM) in either secondary electron or electron beam-induced current (EBIC) regime, using cross sections of the structures with a p-n junction formed in the subsurface region of the active layer. The measurements of defect recombination activity were made using Schottky diodes formed on the active layer surfaces. Parasitic p-n junctions in some samples under a small direct voltage have been observed and the reason for the appearance of such parasitic junctions has been established. Two types of defects with different distribution densities and amplitudes of EBIC contrast have been detected.     

Quantitative X-ray projection microscopy: phase-contrast and multi-spectral imaging

We outline a new approach to X‐ray projection microscopy in a scanning electron microscope (SEM), which exploits phase contrast to boost the quality and information content of images. These developments have been made possible by the combination of a high‐brightness field‐emission gun (FEG)‐based SEM, direct detection CCD technology and new phase retrieval algorithms. Using this approach we have been able to obtain spatial resolution of < 0.2 µm and have demonstrated novel features such as: (i) phase‐contrast enhanced visibility of high spatial frequency image features (e.g. edges and boundaries) over a wide energy range; (ii) energy‐resolved imaging to simultaneously produce multiple quasi‐monochromatic images using broad‐band polychromatic illumination; (iii) easy implementation of microtomography; (iv) rapid and robust phase/amplitude‐retrieval algorithms to enable new real‐time and quantitative modes of microscopic imaging. These algorithms can also be applied successfully to recover object–plane information from intermediate‐field images, unlocking the potentially greater contrast and resolution of the intermediate‐field regime. Widespread applications are envisaged for fields such as materials science, biological and biomedical research and microelectronics device inspection. Some illustrative examples are presented. The quantitative methods described here are also very relevant to projection microscopy using other sources of radiation, such as visible light and electrons.     

Quantitative high-resolution microscopy on a suspended chain of gold atoms

High-resolution electron microscope images of a suspended gold chain were analyzed quantitatively. The images of the suspended gold chain were recorded on videotapes, during in situ observation in an ultra-high-vacuum electron microscope. The chain consisted of four gold atoms free from any substrate. Image simulation verified that the gold chain has a center-top geometry. The simulation also showed that the chain has no glue atoms spinning around the axis of the chain. A spinning gold atom was shown to give 50% of the stationary atom in gray scale. By comparing the noise level of the observed image and the gray scale of single atoms such as C, Si, S, Cu, and Au, we verified that the Si and S atoms have almost twice as large contrast as the noise (peak-to-peak), and C atoms have almost the same contrast as the noise level. The analysis showed it critical to detect a single Si and S atoms.