Application of channel electron multipliers in an electron detector for low‐voltage scanning electron microscopy

An electron detector containing channel electron multipliers was built and tested in the range of low‐voltage scanning electron microscopy as a detector of topographic contrast. The detector can detect backscattered electrons or the sum of backscattered electrons and secondary electrons, with different amount of secondary electrons. As a backscattered electron detector it collects backscattered electrons emitted in a specific range of take‐off angles and in a large range of azimuth angles enabling to obtain large solid collection angle and high collection efficiency. Two arrangements with different channel electron multipliers were studied theoretically with the use of the Monte Carlo method and one of them was built and tested experimentally. To shorten breaks in operation, a vacuum box preventing channel electron multipliers from an exposure to air during specimen exchanges was built and placed in the microscope chamber. The box is opened during microscope observations and is moved to the side of the scanning electron microscope chamber and closed during air admission and evacuation cycles enabling storing channel electron multipliers under vacuum for the whole time. Experimental tests of the detector included assessment of the type of detected electrons (secondary or backscattered), checking the tilt contrast, imaging the spatial collection efficiency, measuring the noise coefficient and recording images of different specimens.     

The effect of subsurface topography on secondary electron images from chromate pretreated aluminium surfaces

In secondary and scanning transmission electron microscopes, secondary electron images of surface films can be dominated by an image derived from electrons back-scattered from the interface between the film and the substrate. The extent of the domination has been established by studying the variation in image obtained using primary beams of different energy and by platinum coating to enhance surface secondary electron emission. Studies of thicker films also established that chemical or structural difference within a film also lead to imaging effects. In general, 5 keV electrons are the most effective in producing subsurface and structural or chemical imaging effects.     

Investigation of quantitative secondary electron imaging of semiconducting polymer materials using environmental scanning electron microscopy

The development of environmental scanning electron microscopy has opened the way for the examination of a wide variety of new sample types that were previously inaccessible to conventional scanning electron microscope techniques. With the advent of such a new methodology comes also the potential for new contrast mechanisms. We investigated the use of environmental scanning electron microscopy on semiconducting organic polymer materials. We observed contrast from these materials in secondary electron images, this contrast being attributed to differences in electron yield due to the polymer's electronic structure. Further study of these materials, and specifically the influence of film thickness on signal, has indicated a significant effect as the secondary electrons move through the sample. Systematic studies such as these are needed for a full understanding of the relationship between electronic properties and signal and, hence, the ability to probe structure–property relationships in greater depth.     

Scanning electron microscope electron detector with a radial type discrete dynode electron multiplier

A discrete dynode electron multiplier with radial flux of electrons was built and tested in the range of low‐voltage scanning electron microscopy as a backscattered electron detector of topographic contrast. The multiplier collects backscattered electron emitted in a specific range of take‐off angles and over the whole azimuth angular range enabling large solid collection angle. Multipliers with different dynode shapes were studied theoretically with the use of the software for particle optics and three assemblies were built and tested experimentally. The gain estimation, assessment of the type of detected electrons (secondary electron or backscattered electron), imaging the spatial collection efficiency and signal‐to‐noise measurements were performed.     

Pushing the resolution limits in cryo electron tomography of biological structures

Cryo electron tomography is a three-dimensional imaging technique that is suitable for imaging snapshots of the structural arrangements of biomolecular complexes and macromolecules, both in vitro and in the context of the cell. In terms of attainable resolution, cryo electron tomographic reconstructions now show resolvable details in the 5-10 nm range, connecting optical microscopy with molecular imaging techniques. In view of the current developments in super-resolution light microscopy and correlative light and electron microscopy, cryo electron tomography will be increasingly important in structural biology as a tool to bridge light microscopy with molecular imaging techniques like NMR, X-ray diffraction and single particle electron microscopy. In cell biology, one goal, often referred to as visual proteomics, is the molecular mapping of whole cells. To achieve this goal and link cryo electron tomography to these high-resolution techniques, increasing the attainable resolution to 2-5 nm is vital. Here, we provide an overview of technical factors that limit the resolution in cryo electron tomography and discuss how during data acquisition and image processing these can be optimized to attain the highest possible resolution. Also, existing resolution measurement approaches and current technological developments that potentially increase the resolution in cryo electron tomography are discussed.     

Zero-loss energy-filtered imaging of frozen-hydrated proteins: model calculations and implications for future developments

Energy-filtered transmission electron microscopes operating in zero-loss mode are used increasingly to study biological material in frozen-hydrated conditions. The contrast enhancement and improved structural resolution obtainable by this method have been studied using Monte-Carlo model calculations for the scattering processes occurring in such samples. Three models representing typical situations have been analysed, each normalized to minimal beam damage. It is shown that for proteins in thin layers of ice an optimal signal-to-noise ratio is achieved in the 80–120-keV electron energy range. For proteins which have to be embedded in thicker ice layers, a considerably higher acceleration voltage is required. In particular, electron energies above 200 keV would be desirable for electron diffraction work on microcrystals.     

A high signal-to-noise ratio toroidal electron spectrometer for the SEM

This paper presents a high signal-to-noise ratio electron energy spectrometer attachment for the scanning electron microscope (SEM), designed to measure changes in specimen surface potential from secondary electrons and extract specimen atomic number information from backscattered electrons. Experimental results are presented, which demonstrate that the spectrometer can in principle detect specimen voltage changes well into the sub-mV range, and distinguish close atomic numbers by a signal-to-noise ratio of better than 20. The spectrometer has applications for quantitatively mapping specimen surface voltage and atomic number variations on the nano-scale.     

Fractal behavior of electron trajectories and spatial distributions of secondary electron birth sites in solids

The fractal behavior of electron scattering in solids, simulated by the Monte Carlo method, has been studied. The trajectories of electrons of 1–10 keV primary energies in Si, Cu, Ag, and Au have been simulated. The Hausdorff dimensions of trajectories of primary electrons, trajectories of primary electrons plus secondary electrons, and spatial distributions of secondary electron birth sites have been determined by the box-counting method. The fractal geometry can be used to describe quantitatively the complexity of the electron scattering process in solids.     

Electron irradiation-induced destruction of carbon nanotubes in electron microscopes

Observations of carbon nanotubes under exposure to electron beam irradiation in standard transmission electron microscope (TEM) and scanning electron microscope (SEM) systems show that such treatment in some cases can cause severe damage of the nanotube structure, even at electron energies far below the approximate 100 keV threshold for knock-on damage displacing carbon atoms in the graphene structure. We find that the damage we observe in one TEM can be avoided by use of a cold finger. This and the morphology of the damage imply that water vapour, which is present as a background gas in many vacuum chambers, can damage the nanotube structure through electron beam-induced chemical reactions. Though, the dependence on the background gas makes these observations specific for the presently used systems, the results demonstrate the importance of careful assessment of the level of subtle structural damage that the individual electron microscope system can do to nanostructures during standard use.     

New technique for in-situ measurement of backscattered and secondary electron yields for the calculation of signal-to-noise ratio in a SEM

The quality of an image generated by a scanning electron microscope is dependent on secondary emission, which is a strong function of surface condition. Thus, empirical formulae and available databases are unable to take into account actual metrology conditions. This paper introduces a simple and reliable measurement technique to measure secondary electron yield (δ) and backscattered electron yield (η) that is suitable for in-situ measurements on a specimen immediately prior to imaging. The reliability of this technique is validated on a number of homogenous surfaces. The measured electron yields are shown to be within the range of published data and the calculated signal-to-noise ratio compares favourably with that estimated from the image.     

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.     

SEM electron channelling patterns as a technique for the characterization of ion-implantation damage

Electron channelling patterns (ECPs) formed in back-scattered images in the scanning electron microscope (SEM) have been used occasionally to confirm surface amorphization during ion implantation. In order to place such observations on a more quantitative basis, the study reported here has explored the variation of ECP appearance with both specimen damage levels (and thus subsurface structures) and SEM accelerating voltage (i.e. sampled depth). Polished and annealed (0001) single crystal sapphire discs were implanted to various damage levels up to both subsurface and full surface amorphization. Damage levels were measured independently by Rutherford back-scattering (RBS). Selected-area ECPs were obtained in a Jeol-840 electron microscope operating over the range 5–40 kV in 5-kV steps. Progressive ECP degradation—in terms of high-order line disappearance—was observed with increasing dose, culminating in total pattern loss when full surface amorphization occurred. However, ECP information could still be obtained from the damaged near-surface material even when a subsurface amorphous layer was present, thus demonstrating the shallow retrieval depth of information from the ECP technique. Indeed, because the spatial distribution of damage from ion implantation is both calculable and measurable, these experiments have also allowed us, for the first time, to explore and demonstrate the shallow sample depths from which the majority of ECP contrast originates (< 150 nm in sapphire at an accelerating voltage of 35 kV), even when the beam penetration is considerable by comparison (～ 5 μm). Furthermore, the way in which this sampled depth varies with SEM accelerating voltage is both demonstrated and shown to be a powerful diagnostic technique for studying the distribution of near-surface structural damage.     

Quantitative secondary ion mass spectrometry imaging of self-assembled monolayer films for electron beam dose mapping in the environmental scanning electron microscope

Fluorinated alkanethiol self-assembled monolayers (SAM) films immobilized on gold substrates have been used as electron-sensitive resists to map quantitatively the spatial distribution of the primary electronbeam scattering in an environmental scanning electron microscope (ESEM). In this procedure, a series of electron dose standards are prepared by exposing a SAM film to electron bombardment in well-defined regions at different levels of electron dose. Microbeam secondary ion mass spectrometry (SIMS) using Cs⁺ bombardment is then used to image the F^- secondary ion signal from these areas. From the reduction in F^- intensity as a function of increasing electron dose, a calibration curve is generated that allows conversion of secondary ion signal to electron dose on a pixel-by-pixel basis. Using this calibration, electron dose images can be prepared that quantitatively map the electron scattering distribution in the ESEM with micrometer spatial resolution. The SIMS imaging technique may also be used to explore other aspects of electron-surface interactions in the ESEM.     

New system for secondary electron detection in variable-pressure scanning electron microscopy

A novel secondary electron detection system combining a two‐stage detector head and a differential pumping system is presented. The detector head consisted of a scintillation Everhart‐Thornley detector and a microsphere plate, separating it from the lower vacuum in the intermediate chamber (below 0.1 mbar). The system was arranged asymmetrically, which should contribute to a lower gas leakage through the plate and a longer life span of the plate. The system offered all the advantages of the scintillator detector in a wide range of gas pressures, from high vacuum to those of the order of 10 mbar, typical of high‐pressure scanning electron microscopy.     

Effect of carbon nanotube structural parameters on field emission properties

Experimental studies were devoted to the effect of structural parameters, i.e., tube diameter and density, on the field electron emission characteristics of carbon nanotubes. Thermal chemical vapor deposition system was employed to synthesize carbon nanotubes. Nanotubes with different diameters and densities were obtained by adjusting the thickness of the iron (Fe) catalyst film. The morphologies of the Fe and carbon nanotube film were characterized by scanning electron microscopy respectively. Further field emission measurement confirmed that the tube diameter and density could significantly affect the electron emission properties of the carbon nanotube. Possible physical reasons for the effect are discussed.     

Resolution in low voltage scanning electron microscopy

As the energy of an electron beam is reduced, the range falls and the secondary electron yield rises. A low voltage scanning electron microscope can therefore, in principle, examine without damage or charging samples such as insulators, dielectrics or beam sensitive materials. This paper investigates the way in which the choice of beam energy affects the spatial resolution of a secondary electron image. It is shown that for samples which are thin compared to the electron range, the edge resolution and contrast in the image improve with increasing beam energy. In samples that are thicker than the electron range, the resolution can be optimized at either high or low energies, but low energy operation will produce images of higher contrast. At an energy of 2 keV or less beam interaction limited resolutions of the order of 3 nm should be possible.     

Effect of electron beam-induced deposition and etching under bias

Electron-beam-induced deposition (EBID) and etching (EBIE) provides a simple way to fabricate or etch submicron or nanoscale structures of various materials in a direct-write (i.e.nonlithographic) fashion. The growth rate or the etch rate are influenced by many factors such as beam energy, beam current, temperature of the substrate material, pressure of the chamber, and geometry of the gas injector etc. The mechanism of EBID and EBIE involves the interaction of the incident electron beam or emitted electron from the target material. The role of these electrons is still not completely understood although the contribution of low energy secondary electrons (SE) has been assumed to be the dominant contributor of EBID and EBIE based on its overlap with the dissociation cross section. We have studied the growth and etching phenomenon under various biasing conditions to investigate how low voltage biasing of the substrate affects secondary electron trajectories and subsequently modifies electron-beam-induced deposition and etching.     

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.     

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.