Reverse Monte Carlo reconstruction algorithm for discrete electron tomography based on HAADF‐STEM atom counting

In this paper, we propose an algorithm to obtain a three‐dimensional reconstruction of a single nanoparticle based on the method of atom counting. The location of atoms in three dimensions has been successfully performed using simulations of high‐angle‐annular‐dark‐field images from only three zone‐axis projections, [110], [310] and [211], for a face‐centred cubic particle. These three orientations are typically accessible by low‐tilt holders often used in high‐performance scanning transmission electron microscopes.     

STEM imaging of thick specimens with off-axis detectors

For scanning transmission electron microscopy (STEM) images obtained with relatively small objective aperture sizes, the contrast of small objects contained within thick specimens may be considerably enhanced by using an off-axis detector aperture situated on the edge of the central beam spot. The effect is demonstrated for both crystalline and amorphous specimens. The effect arises because the detector collects part of the small angle inelastic scattering and is modified by refraction effects for specimens of rapidly changing thickness.     

Quantitation of SEM EBIC and CL signals using Monte Carlo electron-trajectory simulations

A microcomputer Monte Carlo program simulates electron trajectories in solids and describes the distribution of energy deposited throughout the energy-dissipation (electron-hole pair generation) volume. From this distribution, the electron-beam-induced current or cathodoluminescence signal that will be generated can be calculated for the chosen beam conditions in a multilayer specimen of any geometry and compositions. The use of this program is illustrated by applications (1) to simulate curves of cathodoluminescence intensity versus beam energy for fitting to experimental data to evaluate materials and device parameters, (2) to calculate the energy deposited in each layer of a HEMT structure in which electron-beam-induced current studies are in progress, and (3) to the simulation of defect contrast linescan profiles which are compared to experimental observations.     

Scanning electron microscopic observations on deembedded biological tissue sections: Comparison of different fixatives and embedding materials

Conventional plant histological specimens fixed in formalin-acetic acid-alcohol, chromic acid-acetic acid-formaldehyde, or glutaraldehyde-osmium and embedded in either paraffin or plastic are examined as possible rapid methods for providing an alternative image of cellular structure by using scanning electron microscopy. Using the mitotic figures of actively growing onion root tips as a study specimen, the organization of the nucleus and spindle apparatus is reasonably well preserved as compared with isolated mitotic spindles and studies of mitosis in endosperm tissue. Relief of internal structure in this technique is obtained through the coagulant nature of the fixative. Used judiciously, this technique can reveal aspects of the three-dimensional nature of internal tissue structure that may otherwise be difficult to discern.     

Quantification of metallic nanoparticle morphology on TiO2 using HAADF-STEM tomography

Electron tomography is applied to photocatalytic gold/titanium oxide and gold/silver/titanium oxide samples. In order to obtain a tilt series for the electron tomography measurement, high-angle annular dark-field scanning transmission electron microscopy (HAADF-STEM) is used under cryogenic conditions. Dedicated programs have been developed for measuring volume, surface area, thickness distribution and nearest-neighbour distance of metallic nanoparticles on samples. Using these quantification programs, the 3D morphology of gold and silver nanoparticles is accurately characterized. We paid particular attention to the quantitative measurement of surface area. The measurement error of the method and appropriate magnification are defined using spherical nanoparticle models. We measured the 3D morphology of gold nanoparticles supported on titanium oxide (total volume=6.5×10⁵ [nm³], surface area=1.4×10⁵ [nm²], and average nearest-neighbour distance=40 [nm]).     

Imaging thin and thick sections of biological tissue with the secondary electron detector in a field-emission scanning electron microscope

A field-emission scanning electron microscope (FESEM) equipped with the standard secondary electron (SE) detector was used to image thin (70–90 nm) and thick (1–3 μm) sections of biological materials that were chemically fixed, dehydrated, and embedded in resin. The preparation procedures, as well as subsequent staining of the sections, were identical to those commonly used to prepare thin sections of biological material for observation with the transmission electron microscope (TEM). The results suggested that the heavy metals, namely, osmium, uranium, and lead, that were used for postfixation and staining of the tissue provided an adequate SE signal that enabled imaging of the cells and organelles present in the sections. The FESEM was also used to image sections of tissues that were selectively stained using cytochemical and immunocytochemical techniques. Furthermore, thick sections could also be imaged in the SE mode. Stereo pairs of thick sections were easily recorded and provided images that approached those normally associated with high-voltage TEM.     

A new heavy-metal-containing resin for low-temperature embedding and imaging of unstained sections of biological material

A styrene-based, low viscosity, negative-staining resin has been formulated for low-temperature embedding of unstained biological material. The resin, containing 3 atom% tin, permits sample preparations at 243°K, and provides sufficient contrast with bright field conventional transmission electron microscopy (CTEM) and excellent contrast in scanning transmission electron microscopy (STEM) ratio mode.     

Monte Carlo simulation of electron backscattering from compounds with low mean atomic number

This paper reports a Monte Carlo simulation where a single atom scattering model is adopted. The element taking part in each electron-atom interaction is selected on the basis of its contribution eitherto the total elastic cross section or to the electron's mean free path. Both Rutherford and Mott scattering are considered, with the continuous slowing down process of Bethe used to calculate the energy loss to the system. The backscattered electron coefficients show good agreement with experimental results from a large group of low atomic number materials when using a model which selects the scattering atom by its contribution to the whole compound calculated from its atomic fraction of the total elastic cross-section.     

Optimal experimental design of STEM measurement of atom column positions

A quantitative measure is proposed to evaluate and optimize the design of a high-resolution scanning transmission electron microscopy (STEM) experiment. The proposed measure is related to the measurement of atom column positions. Specifically, it is based on the statistical precision with which the positions of atom columns can be estimated. The optimal design, that is, the combination of tunable microscope parameters for which the precision is highest, is derived for different types of atom columns. The proposed measure is also used to find out if an annular detector is preferable to an axial one and if a C_s-corrector pays off in quantitative STEM experiments. In addition, the optimal settings of the STEM are compared to the Scherzer conditions for incoherent imaging and their dependence on the type of object is investigated.     

Optimization of STEM tomography acquisition — A comparison of convergent beam and parallel beam STEM tomography

In this paper two imaging modes in a state-of-the-art scanning transmission electron microscope (STEM) are compared: conventional STEM with a convergent beam (referred to as nanoprobe) and STEM with a parallel beam (referred to as microprobe). The effect and influence of both modes with respect to their depth of field are investigated. Tomograms of a human white blood cell (hemophagocytes) are acquired, aligned, and evaluated. It is shown that STEM using a parallel beam produces tomograms with fewer distortions and artifacts that allows resolving finer features. Microprobe STEM tomography is advantageous especially in life science, when semi-thin sections (approximately 0.5 μm thick) of biological samples are imaged at relatively low magnification with a large field of view.     

Optimal strategies for imaging thick biological specimens: exit wavefront reconstruction and energy-filtered imaging

In transmission electron microscopy (TEM) of thick biological specimens, the relationship between the recorded image intensities and the projected specimen mass density is distorted by incoherent electron–specimen interactions and aberrations of the objective lens. It is highly desirable to develop a strategy for maximizing and extracting the coherent image component, thereby allowing the projected specimen mass density to be directly related to image intensities. For this purpose, we previously used exit wavefront reconstruction to understand the nature of image formation for thick biological specimens in conventional TEM. Because electron energy-loss filtered imaging allows the contributions of inelastically scattered electrons to be removed, it is potentially advantageous for imaging thick, biological samples. In this paper, exit wavefront reconstruction is used to quantitatively analyse the imaging properties of an energy-filtered microscope and to assess its utility for thick-section microscopy. We found that for imaging thick biological specimens (> 0.5 μm) at 200 keV, only elastically scattered electrons contribute to the coherent image component. Surprisingly little coherent transfer was seen when using energy-filtering at the most probable energy loss (in this case at the first plasmon energy-loss peak). Furthermore, the use of zero-loss filtering in combination with exit wavefront reconstruction is considerably more effective at removing the effects of multiple elastic and inelastic scattering and microscope objective lens aberrations than either technique by itself. Optimization of the zero-loss signal requires operation at intermediate to high primary voltages (> 200 keV). These results have important implications for the accurate recording of images of thick biological specimens as, for instance, in electron microscope tomography.     

High-angle triple-axis specimen holder for three-dimensional diffraction contrast imaging in transmission electron microscopy

Electron tomography requires a wide angular range of specimen-tilt for a reliable three-dimensional (3D) reconstruction. Although specimen holders are commercially available for tomography, they have several limitations, including tilting capability in only one or two axes at most, e.g. tilt-rotate. For amorphous specimens, the image contrast depends on mass and thickness only and the single-tilt holder is adequate for most tomographic image acquisitions. On the other hand, for crystalline materials where image contrast is strongly dependent on diffraction conditions, current commercially available tomography holders are inadequate, because they lack tilt capability in all three orthogonal axes needed to maintain a constant diffraction condition over the whole tilt range. We have developed a high-angle triple-axis (HATA) tomography specimen holder capable of high-angle tilting for the primary horizontal axis with tilting capability in the other (orthogonal) horizontal and vertical axes. This allows the user to trim the specimen tilt to obtain the desired diffraction condition over the whole tilt range of the tomography series. To demonstrate its capabilities, we have used this triple-axis tomography holder with a dual-axis tilt series (the specimen was rotated by 90° ex-situ between series) to obtain tomographic reconstructions of dislocation arrangements in plastically deformed austenitic steel foils.     

Monte Carlo simulation of secondary electron images for real sample structures in scanning electron microscopy

Monte Carlo simulation methods for the study of electron beam interaction with solids have been mostly concerned with specimens of simple geometry. In this article, we propose a simulation algorithm for treating arbitrary complex structures in a real sample. The method is based on a finite element triangular mesh modeling of sample geometry and a space subdivision for accelerating simulation. Simulation of secondary electron image in scanning electron microscopy has been performed for gold particles on a carbon substrate. Comparison of the simulation result with an experiment image confirms that this method is effective to model complex morphology of a real sample.     

A novel method of Z-contrast imaging in STEM applied to double-labelling

A novel method of Z-contrast imaging in the scanning transmission electron microscope (STEM) is presented. The technique relies on the element dependence of the angular distribution of the scattered electrons, and is realized with a detector consisting of a set of concentric rings. It is possible to discriminate 9-nm colloidal gold and silver specifically distributed on thin sections. In addition to this practical work, numerical evaluations are used to assess the method. With two smaller markers, this approach will be useful in discriminating closely spaced antigenic sites when steric hindrance occurs with double-labelling using probes of different sizes.     

Use of Monte Carlo modeling for interpreting scanning electron microscope linewidth measurements

A scanning electron microscope (SEM) can be used to measure the dimensions of the microlithographic features of integrated circuits. However, without a good model of the electron-beam/specimen interaction, accurate edge location cannot be obtained. A Monte Carlo code has been developed to model the interaction of an electron beam with one or two lines lithographically produced on a multilayer substrate. The purpose of the code is to enable one to extract the edge position of a line from SEM measurements. It is based on prior codes developed at the National Institute of Standards and Technology, but with a new formulation for the atomic scattering cross sections and the inclusion of a method to simulate edge roughness or rounding. The code is currently able to model the transmitted and backscattered electrons, and the results from the code have been applied to the analysis of electron transmission through gold lines on a thin silicon substrate, such as is used in an x-ray lithographic mask. Significant reductions in backscattering occur because of the proximity of a neighboring line.     

Monte Carlo study for correcting the broadened line-scan profile in scanning electron microscopy

Line-scan profile is always broadened due to the probe shape of the primary electron (PE) beam in scanning electron microscopy (SEM), which leads to an inaccurate dimension metrology. Currently, the effective electron beam shape (EEBS) is suggested as the broadening function to overcome this issue for theoretical analysis, rather than the widely used Gaussian profile. However, EEBS is almost impossible to be acquired due to it strongly depends on both the sample topography and the electron beam focusing condition, which makes it is impossible to be applied in practical analysis. Taking the case of gate linewidth measurement, an approach is proposed to find a best-fit traditional Gaussian profile, which can optimally replace the EEBS in the case of the same sample structure and experimental condition for construction of a database of the parameter in traditional Gaussian profile. This approach is based on the use of the ideal and broadened line-scan profiles which are both obtained from Monte Carlo (MC) simulation, but respectively by an ideal and a focusing incident electron beam model. The expected value of parameter can be obtained through deconvoluting (here using a maximum-entropy algorithm) the broadened line-scan profile then fitting it to the ideal profile. Experimenters can benefit from this database to obtain true line-scan profiles for accurate gate linewidth measurement. This work should prove useful for samples of other structures and be an extension of the database in the future.     

Lateral sensitivity in electron probe microanalysis studied by Monte Carlo simulations involving fluorescence enhancements

In electron probe microanalysis, secondary fluorescence can occur leading to an increase of the volume analysed, degrading the lateral resolution of this technique. An adequate knowledge of the interaction volumes from where the different signals of interest are detected is determinant to estimate the minimum size of the zone that can be characterized. In this work, the size of the signal source volume is surveyed for a wide set of samples at different beam energies. To this aim, the PENELOPE software package was chosen to run Monte Carlo simulations for several experimental situations in order to produce the various lateral radiation distributions of interest. A comparison between the interaction volumes of the different signals was performed by taking into account the different fluorescence enhancement possibilities. An unexpected behaviour was found in the particular cases of aluminium and alumina, where the secondary photons signal exhibits a decreasing trend up to certain beam energy (～17 keV); this implies that lower beam energies may degrade the lateral resolution of the technique in these materials.     

Monte Carlo simulation of secondary electron and backscattered electron images in scanning electron microscopy for specimen with complex geometric structure

A new Monte Carlo technique for the simulation of secondary electron (SE) and backscattered electron (BSE) of scanning electron microscopy (SEM) images for an inhomogeneous specimen with a complex geometric structure has been developed. The simulation is based on structure construction modeling with simple geometric structures, as well as on the ray-tracing technique for correction of electron flight-step-length sampling when an electron trajectory crosses the interface of the inhomogeneous structures. This correction is important for the simulation of nanoscale structures of a size comparable with or even less than the electron scattering mean free paths. The physical model for electron transport in solids combines the use of the Mott cross section for electron elastic scattering and a dielectric function approach for electron inelastic scattering, and the cascade SE production is also included.     

基于蒙特卡罗模拟的NaI，BGO，LSO，GSO，YSO闪烁晶体对比研究

发射断层成像系统中大都采用闪烁晶体与光电倍增管相耦合组成的γ射线探测器。闪烁晶体的光收集效率直接影响着探测器的能量、时间和空间分辨力。为了设计更好的探测器,利用M onte Carlo方法对N aI,BGO,LSO,GSO,YSO闪烁晶体进行了对比研究,分析比较了不同表面处理、晶体尺寸,外界反射材料对光收集效率的影响。模拟结果表明:入射面为粗糙面,其余为抛光面同时外层包装上高反射率的材料可得到最大的光输出,其中YSO表现出最优的光输出性能。晶体的几何尺寸对得到高的光输出也起着非常重要的作用。