Low voltage scanning electron microscopy

The scanning electron microscope (SEM) is usually operated with a beam voltage, V₀, in the range of 10–30 kV, even though many early workers had suggested the use of lower voltages to increase topographic contrast and to reduce specimen charging and beam damage. The chief reason for this contradiction is poor instrumental performance when V₀=1–3 kV, The problems include low source brightness, greater defocusing due to chromatic aberration greater sensitivity to stray fields, and difficulty in collecting the secondary electron signal. Responding to the needs of the semiconductor industry, which uses low V₀ to reduce beam damage, considerable efforts have been made to overcome these problems. The resulting equipment has greatly improved performance at low kV and substantially removes the practical deterrents to operation in this mode. This paper reviews the advantages of low voltage operation, recent progress in instrumentation and describes a prototype instrument designed and built for optimum performance at 1 kV. Other limitations to high resolution topographic imaging such as surface contamination, the de-localized nature of the inelastic scattering event and radiation damage are also discussed.     

Simulations of scanning electron microscopy imaging and charging of insulating structures

We present a three‐dimensional simulation of scanning electron microscope (SEM) images and surface charging. First, the field above the sample is calculated using Laplace's equation with the proper boundary conditions; then, the simulation algorithm starts following the electron trajectory outside the sample by using electron ray tracing. When the electron collides with the specimen, the algorithm keeps track of the electron inside the sample by simulating the electron scattering history with a Monte Carlo code. During this phase, secondary and backscattered electrons are emitted to form an image and primary electrons are absorbed; therefore, a charge density is formed in the material. This charge density is used to recalculate the field above and inside the sample by solving the Poisson equation with the proper boundary conditions. Field equation, Monte Carlo scattering simulation, and electron ray tracing are therefore integrated in a self‐consistent fashion to form an algorithm capable of simulating charging and imaging of insulating structures. To maintain generality, this algorithm has been implemented in three dimensions. We shall apply the so‐defined simulation to calculate both the global surface voltage and local microfields induced by the scanning beam. Furthermore, we shall show how charging affects resolution and image formation in general and how its characteristics change when imaging parameters are changed. We shall address magnification, scanning strategy, and applied field. The results, compared with experiments, clearly indicate that charging and the proper boundary conditions must be included in order to simulate images of insulating features. Furthermore, we shall show that a three‐dimensional implementation is mandatory for understanding local field formation.     

Quantitative electron spectroscopic imaging studies of microelectronic metallization layers

The combination of focused ion beam (FIB) sample preparation and quantitative electron spectroscopic imaging is an ideal tool for the investigation of layered structures used in microelectronic metallization schemes. In the present work, Si₃N₄/Cu/Si₃N₄/SiO₂/Si and Al/TiN/Ti/SiO₂/Si metallization layers produced by physical vapour deposition are investigated. We apply series of energy filtered images in the low loss region for a mapping of the sample thickness which makes it possible to refine the parameters of the FIB process. We also show how series of energy filtered images in the core loss region can be used to obtain elemental distribution images and chemical bonding information on these samples on a nanometre scale. For materials with a small grain size and/or a strong variation in Bragg orientation, the intensity distribution of the elemental map is strongly influenced by the superimposed Bragg contrast. This detrimental effect can be reduced greatly by using hollow cone illumination, as is demonstrated for polycrystalline Cu. One striking feature observed in Cu layers prepared with FIB is strong, regularly arranged contrast variations caused by subsurface defects in the Cu grains. We suppose that these defects are a consequence of a strong interaction of Ga atoms from the FIB with Cu.     

Recent developments and new strategies in scanning electron microscopy*

In addition to improvements in lateral resolution in scanning electron microscopy, recent developments of interest here concern extension of the incident beam energy, E₀, over two decades, from ≈ 20 keV to ≈ 0.1–0.5 keV and the possibility of changing the take-off emission, α, of detected secondary electrons. These two degrees of freedom for image acquisition permit a series of images of the same field of view of a specimen to be obtained, each image of the series differing from the others in some aspect. The origins of these differences are explored in detail and they are tentatively interpreted in terms of the change in the secondary electron emission yield δ vs. E₀, δ = f(E₀), and also of the change in δ vs. α, ∂δ/∂α. Various origins for the chemical contrast and topographic contrast have been identified. Illustrated by correlating a secondary electron image and a backscattered electron image, use of the scatter diagram technique facilitates image comparison. The difference between the lateral resolution and the size of the minimum detectable detail is outlined to avoid possible errors in nanometrology. Some aspects related to charging are also considered and possible causes of contrast reversal are suggested. Finally, the suggested strategy consists of the acquisition of various images of a given specimen by changing one parameter: primary beam energy and take-off angle for conductive specimens; working distance or beam intensity for high-resolution experiments; scanning frequency for insulating specimens.     

A systematic investigation of the charging effect in scanning electron microscopy for metal nanostructures on insulating substrates

Scanning electron microscopy is perhaps the most important method for investigating and characterizing nanostructures. A well‐known challenge in scanning electron microscopy is the investigation of insulating materials. As insulating materials do not provide a path to ground they accumulate charge, evident as image drift and image distortions. In previous work, we have seen that sample charging in arrays of metal nanoparticles on glass substrates leads to a shrinkage effect, resulting in a measurement error in the nanoparticle dimension of up to 15% at 10 kV and a probe current of 80 ± 10 pA. In order to investigate this effect in detail, we have fabricated metal nanostructures on insulating borosilicate glass using electron beam lithography. Electron beam lithography allows us to tailor the design of our metal nanostructures and the area coverage. The measurements are carried out using two commonly available secondary electron detectors in scanning electron microscopes, namely, an InLens‐ and an Everhart–Thornley detector. We identify and discriminate several contributions to the effect by varying microscope settings, including the size of the aperture, the beam current, the working distance and the acceleration voltage. We image metal nanostructures of various sizes and geometries, investigating the influence of scan‐direction of the electron beam and secondary electron detector used for imaging. The relative measurement error, which we measure as high as 20% for some settings, is found to depend on the acceleration voltage and the type of secondary electron detector used for imaging. In particular, the Everhart–Thornley detectors lower sensitivity to SE₁ electrons increase the magnitude of the shrinkage of up to 10% relative to the InLens measurements. Finally, a method for estimating charge balance in insulating samples is presented.     

Calibration of an electron back-scattering pattern set-up

The determination of lattice orientations from electron back-scattering patterns (EBSPs) in a scanning electron microscope (SEM) requires accurate knowledge of the position of the pattern centre and the source point to screen distance. This paper outlines a new procedure that enables the determination of these parameters for any given set-up of the EBSP/SEM system. The calibration procedure simply requires the positions and indices of at least four poles in a pattern obtained from an arbitrary specimen, and eliminates the need for standard specimens or special attachments to the EBSP/SEM system. The pattern centre is shown to be located with a precision of approximately 0·5° and the source point to screen distance can be determined with a relative precision of approximately 0·5%.     

A mirror electron microscope for surface analysis

The principle of mirror microscopy has been adapted to provide a relatively low resolution surface microscope (<1000 ×), a large transfer width low energy electron diffractometer and a photoelectron analyser in k_|| space. A focused electron beam of ? 10 kV is decelerated through a Johansson lens, reflected in front of the sample and reaccelerated back through the lens to produce an electron image over a field of view of a few microns. The image can be interpreted as a micrograph of work function variations on the surface if other effects (geometry, magnetic field) are uniform. In the LEED mode, diffracted beams virtually retain their positions on the screen over the whole impact energy range used (0.160 V). Secondary electrons are preferentially focused around the lens-gun electro-optic axis, thus effectively filtering them out from the diffraction pattern. The design has an inherently large coherence length, of up to 10⁴ Å. Photoelectrons can similarly be imaged in k_|| space on the detector plane. The addition of energy filtering at the screen allows the two-dimensional Fermi surface to be imaged.     

Nanobeam electron diffraction and high resolution imaging analysis of InN films grown on sapphire

A JEOL JEM-3000F field emission, analytical, high-resolution transmission electron microscope (HRTEM) was used to study InN films grown on sapphire substrates. It was found that, while the InN films maintained the hexagonal (wurtzite) structure, InN nanodomains with a cubic (zincblende) structure were also formed in the films. Nanobeam electron diffraction techniques were applied for identification of the cubic phase. The identification of the cubic InN was confirmed by HRTEM structural imaging. The cubic InN nanodomains are 3-10 nm in diameter, and are orientated in two different orientations with their [110](cubic) and [110](cubic) axes parallel to each other and their (111)(cubic) planes parallel to the (0001)(hex) plane of the hexagonal InN.     

Studies of supported metal catalysts using high resolution secondary electron imaging in a STEM

An additional technique for use in the characterization of catalysts by electron microscopy is presented. High resolution secondary electron images obtained in a VG HB501 scanning transmission electron microscope have been used to study the surface topography of catalysts consisting of small metal particles on high surface area carbon supports. Surface features down to nanometre dimensions can be seen, allowing the examination of micropores in the support as well as larger pore structures. The results are compared with pore size distributions determined by gas adsorption methods, and are shown to yield valuable additional information. In addition, the method in principle allows examination of the locations of small metal catalyst particles on the support.     

3D visualization and quantification of rat cortical bone porosity using a desktop micro-CT system: a case study in the tibia

Although micro-computed tomography (micro-CT) has become the gold standard for assessing the 3D structure of trabecular bone, its extension to cortical bone microstructure has been relatively limited. Desktop micro-CT has been employed to assess cortical bone porosity of humans, whereas that of smaller animals, such as mice and rats, has thus far only been imaged using synchrotron-based micro-CT. The goal of this study was to determine if it is possible to visualize and quantify rat cortical porosity using desktop micro-CT. Tibiae (n = 10) from 30-week-old female Sprague-Dawley rats were imaged with micro-CT (3 μm nominal resolution) and sequential ground sections were then prepared. Bland-Altman plots were constructed to compare per cent porosity and mean canal diameter from micro-CT (3D) versus histology (2D). The mean difference or bias (histology-micro-CT; ±95% confidence interval) for per cent porosity was found to be -0.15% (±2.57%), which was not significantly different from zero (P= 0.720). Canal diameter had a bias (±95% confidence interval) of -5.73 μm (±4.02 μm) which was found to be significantly different from zero (P < 0.001). The results indicated that cortical porosity in rat bone can indeed be visualized by desktop micro-CT. Quantitative assessment of per cent porosity provided unbiased results, whereas direct analysis of mean canal diameter was overestimated by micro-CT. Thus, although higher resolution, such as that available from synchrotron micro-CT, may ultimately be required for precise geometric measurements, desktop micro-CT--which is far more accessible--is capable of yielding comparable measures of porosity and holds great promise for assessment of the 3D arrangement of cortical porosity in the rat.     

Reduction of charging effects using vector scanning in the scanning electron microscope.

We describe a vector scanning system to reduce charging effects during scanning electron microscope (SEM) imaging. The vector scan technique exploits the intrinsic charge decay mechanism of the specimen to improve imaging conditions. We compare SEM images obtained by conventional raster scanning versus vector scanning to demonstrate that vector scanning successfully reduces specimen-charging artifacts.     

Cryoprotection in electron microscopy

A set of experiments has been undertaken to resolve questions about the cryoprotection factor afforded by cooling specimens in the electron microscope. Two specimens, an n-alkane (paraffin) and bacterial purple membrane, were prepared and distributed to several laboratories. Radiation damage studies were carried out at these laboratories independently, but following a standard protocol as closely as possible. While it was not possible to make all measurements at all of the laboratories, sufficient data have been obtained to establish a general reproducibility in the overall features of the cryoprotection effect from room temperature to near 4.2 K.     

Post‐thinning using Ar ion‐milling system for transmission electron microscopy specimens prepared by focused ion beam system

We investigate Ar ion‐milling rates and Ga‐ion induced damage on sample surfaces of Si and GaAs single crystals prepared by focused ion beam (FIB) method for transmission electron microscopy observation. The convergent beam electron diffraction technique with Bloch simulation is used to measure the thickness of the Ar‐ion milled samples to calculate the milling rates of Si and GaAs single crystals. The measurement shows that an amorphous layer is formed on the sample surface and can be removed by further Ar‐ion milling. In addition, the local symmetry breaking induced by FIB is investigated using quantitative symmetry measurement. The FIBed‐GaAs sample shows local symmetry breaking after FIB milling, although the FIBed‐Si sample has no considerable symmetry breaking.     

Diffraction patterns of artificial two-dimensional crystals synthesized in situ in an environmental scanning transmission electron microscope

In this study, we demonstrated the use of electron‐beam‐induced deposition for synthesis of artificial two‐dimensional crystals with an in situ scanning transmission electron microscope. The structures were deposited from W(CO)₆ in an environmental scanning transmission electron microscope on a 30‐nm‐thick Si₃N₄ substrate. We present clear electron beam diffraction patterns taken from those structures. The distance between the diffraction peaks corresponded to the dot spacing in the self‐made surface crystal. We propose using these arrays of dots as anchor points for making artificial crystals for diffraction analysis of weakly scattering or beam‐sensitive molecules such as proteins.     

Comparison of ion beam sputtered chromium and iridium films for electron microscopy applications

The structure and composition of thin, conductive metallic films of chromium and iridium that are typical of the coatings used for electron microscopy is described. The purpose of this study was to determine the grain size and composition of the films deposited, with thicknesses of 1 nm, 2.5 nm, and 5 nm, onto amorphous carbon films using ion beam sputtering with argon as the sputtering gas. A comparison between chromium films deposited under conditions of liquid nitrogen (LN) trapping or of no trapping revealed slight differences in their microstructure. As expected, the grain size of the films increased with the thickness, and the average grain sizes varied between 10 and 25 nm. Grain size was also found to depend on the source of ion beam energy; the correlation between grain size and beam energy was more pronounced in the iridium films than in the chromium films. This effect was greater when the deposition chamber was not LN trapped. As the ion beam energy increased from 18 W to 24 W, there was a corresponding increase in grain size in some of the films. Although transmission electron diffraction analysis indicated the presence of about 5% chromium oxide in the chromium films, no oxide was detected in the iridium films.     

In situ observation of oxygen deficiency occurring during electron irradiation in high Tc superconductor YBa2Cu3Oy

The decreasing process of oxygen in YBa₂Cu₃O _y is investigated through high resolution electron microscopy (HREM) and convergent beam electron diffraction (CBED). Measurements of the axial length in HREM images show that oxygen content y decreases faster near a twin boundary than at the inner part of a twin lamella. The transformation from an orthorhombic to a tetragonal phase starts at a twin boundary and the transformed region propagates to an inner region of lamella. Lattice strains are observed near boundaries between transformed and non-transformed regions. The transformation is almost complete within 30 s during observation of HREM images at 400 kV and at room temperature. A value of y was quantitatively measured by analysing observed intensities of energy-filtered CBED patterns with the dynamical theory. The value of y decreases from 6.9 to 6.5 when 200 kV electrons are irradiated for 160 s in a microscope at 108 K. More precise analysis of the intensities provides information on charge distribution along the c -axis as well as local oxygen content at a spatial resolution of several nanometres.     

Investigation and use of plasmon losses in energy-filtering transmission electron microscopy

The electron spectroscopic imaging (ESI), diffraction (ESD) and different types of electron energy-loss spectroscopy (EELS) modes in an energy-filtering transmission electron microscope can all be used for the investigation and analytical use of plasmon losses. Shifts of plasmon losses caused by differences in composition can be detected with an accuracy of 0.1 eV by parallel-recorded EELS (PEELS). The dispersion of plasmon losses and the cut-off angle θ_c can be observed by angle-dispersive EELS and by recording spectra at different scattering angles θ. ESD patterns with a selected energy window width of 1 eV enable the dispersion and its anisotropy to be imaged by characteristic intensity distributions between the primary beam and the first Bragg diffracted beams. The ESI mode can be used for the selective imaging of precipitates and for the investigation of the excitation volume of plasmons in small particles.     

Use of backscattered electron detector arrays for forming backscattered electron images in the scanning electron microscope

The backscattered electron (BSE) signal in the scanning electron microscope (SEM) can be used in two different ways. The first is to give a BSE image from an area that is defined by the scanning of the electron beam (EB) over the surface of the specimen. The second is to use an array of small BSE detectors to give an electron backscattering pattern (EBSP) with crystallographic information from a single point. It is also possible to utilize the EBSP detector and computer-control system to give an image from an area on the specimen--for example, to show the orientations of the grains in a polycrystalline sample ("grain orientation imaging"). Some further possibilities based on some other ways for analyzing the output from an EBSP detector array, are described.     

Fundamental problems of imaging subsurface structures in the backscattered electron mode in scanning electron microscopy

In‐depth imaging of subsurface structures in scanning electron microscopy (SEM) is usually obtained by detecting backscattered electrons (BSE). For a layer‐by‐layer imaging in BSE microtomography, it is preferable to use an energy filtering of BSE. A simple approach is used to estimate the contrast by using backscattering coefficients of bulk materials and the maximum escape depths of the BSE. The contrast obtained by BSE energy filtering is about twice that of the standard BSE method by varying the acceleration voltage. The contrast decreases with increasing information depth. The information depth is about four times smaller than the electron range. The transmission of the spectrometer influences the minimum current of the order of 10^?8 A that is needed to get a contrast of 1%, for example.     

Analytical transmission electron microscopy and electron diffraction for characterization of multiphase Ni-P-Ti surface layer on Ti-6Al-4V alloy

The microstructure, chemical and phase composition of the hard Ni‐P‐Ti layer formed on the Ti‐6Al‐4V alloy after duplex surface treatment were investigated by light microscopy, X‐ray diffraction, scanning electron microscopy and analytical/high‐resolution transmission electron microscopy. These investigations showed that the improved mechanical and tribological properties of the surface‐treated alloy were related to the presence of a multilayered microstructure containing several phases from the Ni‐Ti‐P‐Al system.