Quantitative valence plasmon mapping in the TEM: viewing physical properties at the nanoscale

Using a series of graphitising carbons heat treated at different temperatures, the peak position of the bulk (pi+sigma) plasmon was measured using electron energy loss spectroscopy and observed to shift between 22 and 27eV. Experimental data is presented and discussed showing the effects of the collection conditions and sample orientation upon the observed spectra.We present an empirical technique by which quantitative energy filtered transmission electron microscopy (EFTEM) maps with two energy windows selected in the plasmon region can be readily acquired and processed, the results of which may be interpreted as graphitisation maps and subsequently physical property maps.An experimentally established resolution of approximately 1.6nm makes this technique a very useful tool with which to examine nanoscale properties in microstructural regions of interest in TEM specimens such as fibre/matrix interfaces within carbon-carbon composites, multi-walled carbon nanotubes and graphitic inclusions in carbon steels.Also presented is data demonstrating the unsuitability of pi(*)-related chemical EFTEM maps in both the low-loss region and at the carbon K ionisation edge for mapping bonding in such highly anisotropic media due to the strong orientation dependence of the intensity of the transitions involved. This is followed by suggestions for wider application of the plasmon mapping technique within systems other than those based upon carbon.     

Quantitative comparison of energy-filtering transmission electron microscopy and atom probe tomography

Whereas transmission electron microscopy (TEM) is a well established method for the analysis of thin film structures down to the sub-nanometer scale, atom probe tomography (APT) is less known in the microscopy community. In the present work, local chemical analysis of sputtered Fe/Cr multilayer structures was performed with energy-filtering transmission electron microscopy (EFTEM) and APT. The single-layer thickness was varied from 1 to 6 nm in order to quantify spatial resolution and chemical sensitivity. While both the methods are able to resolve the layer structure, even at 2 nm thickness, it is demonstrated that the spatial resolution of the APT is about a factor of two, higher in comparison with the unprocessed EFTEM data. By calculating the influence of the instrumental parameters on EFTEM images of model structures, remaining interface roughness is indicated to be the most important factor that limits the practical resolution of analytical TEM.     

Energy-filtering TEM at high magnification: spatial resolution and detection limits

Energy-filtering TEM (EFTEM) has turned out to be a very efficient and rapid tool for the chemical characterization of a specimen on a nanometer and even subnanometer length scale. Especially, the detection and measurement of very thin layers has become a great application of this technique in many materials science fields, e.g. semiconductors and hard disk technology. There, the reliability of compositional profiles is an important issue. However, the experimentally obtainable spatial resolution strongly influences the appearance of a thin layer in an EFTEM image, when dimensions reach subnanometer levels, which mainly leads to a broadening of the layer in the image. This fact has to be taken into account, when measuring the thickness of such a thin layer. Additionally, the convolution decreases contrast which makes the layer less visible in the image and finally determines the detection limit.In this work we present a systematic study on specifically designed Mn/PdMn multilayer test specimens to explore the practical aspects of spatial resolution and detection limits in EFTEM. Although specific to the ionization edges used, we will present general conclusions about the practical limitations in terms of EFTEM spatial resolution. Additionally, work will be shown about low energy-loss imaging of thin oxide layers, where delocalization is the main factor responsible for broadening.     

Comparison of EFTEM and STEM EELS plasmon imaging of gold nanoparticles in a monochromated TEM

We present and compare two different imaging techniques for plasmonic excitations in metallic nanoparticles based on high energy-resolution electron energy-loss spectroscopy in a monochromated transmission electron microscope. We demonstrate that a recently developed monochromated energy-filtering (EFTEM) approach can be used in addition to the well established scanning technique to directly obtain plasmon images in the energy range below 1 eV. The EFTEM technique is described in detail, and a double experiment performed on the same, triangular gold nanoparticle compares equivalent data acquired by both techniques, respectively.     

Energy-filtering TEM and electron energy-loss spectroscopy of double structure tabular microcrystals of silver halide emulsions

Composite Ag(Br,I) tabular microcrystals of photographic emulsions were studied by the combination of energy-filtering electron microscopy (EFTEM) and electron energy-loss spectroscopy (EELS) in conjunction with energy-dispersive X-ray (EDX) microanalysis. The contrast tuning under the energy-filtering in the low-loss region was used to observe more clearly edge and random dislocations, {11⁻1} stacking faults in the grain shells parallel to {11⁻2} edges and bend and edge contours. Electron spectroscopic diffraction patterns revealed numerous extra reflections at commensurate positions in between the Bragg reflections and diffuse honeycomb contours; these were assigned to the number of defects in the shell region parallel to the grain edges and polyhedral clusters of interstitial silver cations, respectively. Inner-shell excitation bands of silver halide were detected and confirmed by EDX analyses, i.e. the Ag N_2,3 edge at 62 eV (probably overlapped with the weak I N_4,5 edge at 52 eV and the Br M_4,5 edge at 70 eV), the I M_4,5 edge at about 620 eV, and the Br L_2,3 edge at about 1550 eV energy losses. Energy-loss near-edge structure of the Ag M_4,5 edge at about 367 eV energy losses and low-loss fine structure arisen as a result of interband transitions and excitons, possibly superimposed with many electron effects, have been revealed. The crystal thickness was determined by a modified EELS log-ratio technique in satisfactory agreement with measurements on grain replicas.     

Toward quantitative core-loss EFTEM tomography

Core-loss EFTEM tomography provides three-dimensional structural and chemical information. Multiple inelastic scattering occurring in thick specimens as well as orientation-dependent diffraction contrast due to multiple elastic scattering, however, often limit its applications. After demonstrating the capability of core-loss EFTEM tomography to reconstruct just a few monolayers thin carbon layer covering a Fe catalyst particle we discuss its application to thicker samples. We propose an approximate multiple-scattering correction method based on the use of zero-loss images and apply it successfully to copper whiskers, providing a significant improvement of the reconstructed 3D elemental distribution. We conclude this paper by a general discussion on experimental parameters affecting the accuracy of EFTEM 3D elemental mapping.     

Quantitative EFTEM mapping of near physiological calcium concentrations in biological specimens

Although electron energy-loss spectroscopy (EELS) in the scanning transmission electron microscope (STEM) provides high sensitivity for measuring the important element, calcium, in biological specimens, the technique has been difficult to apply routinely, because of long acquisition times required. Here we describe a refinement of the complementary analytical technique of energy-filtered transmission electron microscopy (EFTEM), which enables rapid imaging of large cellular regions and measurement of calcium concentrations approaching physiological levels. Extraction of precise quantitative information is possible by averaging large numbers of pixels that are contained in organelles of interest. We employ a modified two-window approach in which the behavior of the background signal in the EELS spectrum can be modeled as a function of specimen thickness t expressed in terms of the inelastic mean free path λ. By acquiring pairs of images, one above and one below the Ca L_2,3 edge, together with zero-loss and unfiltered images, which are used to determine a relative thickness (t/λ) map, it is possible to correct the Ca L_2,3 signal for plural scattering. We have evaluated the detection limits of this technique by considering several sources of systematic errors and applied this method to determine mitochondrial total calcium concentrations in freeze-dried cryosections of rapidly frozen stimulated neurons. By analyzing 0.1 μm² areas of specimen regions that do not contain calcium, it was found that the standard deviation in the measurement of Ca concentrations was about 20 mmol/kg dry weight, corresponding to a Ca:C atomic fraction of approximately 2×10⁻⁴. Calcium concentrations in peripheral mitochondria of recently depolarized, and therefore stimulated and Ca loaded, frog sympathetic neurons were in reasonable agreement with previous data.     

Imaging Si nanoparticles embedded in SiO(2) layers by (S)TEM-EELS

Fabrication of systems in which Si nanoparticles are embedded in a thin silica layer is today mature for non-volatile memory and opto-electronics applications. The control of the different parameters (position, size and density) of the nanoparticles population is a key point to optimize the properties of such systems. A review of dedicated transmission electron microscopy (TEM) methods, which can be used to measure these parameters, is presented with an emphasis on those relying on electron energy-loss spectroscopy (EELS). Defocused bright-field imaging can be used in order to determine topographic information of a whole assembly of nanoparticles, but it is not efficient for looking at individual nanoparticles. High-resolution electron imaging or dark-field imaging can be of help in the case of crystalline particles but they always provide underestimated values of the nanocrystals population. EELS imaging in the low-energy-loss domain around the Si plasmon peak, which gives rise to strong signals, is the only way to visualize all Si nanoparticles within a silica film and to perform reliable size and density measurements. Two complementary types of experiments are investigated and discussed more extensively: direct imaging with a transmission electron microscope equipped with an imaging filter (EFTEM) and indirect imaging from spectrum-imaging data acquired with a scanning transmission electron microscope equipped with a spectrometer (STEM-PEELS). The direct image (EFTEM) and indirect set of spectra (STEM-PEELS) are processed in order to deliver images where the contribution of the silica matrix is minimized. The contrast of the resulting images can be enhanced with adapted numerical filters for further morphometric analysis. The two methods give equivalent results, with an easier access for EFTEM and the possibility of a more detailed study of the EELS signatures in the case of STEM-PEELS. Irradiation damage in such systems is also discussed.     

Achieving sub-nanometre particle mapping with energy-filtered TEM

A combination of state-of-the-art instrumentation and optimized data processing has enabled for the first time the chemical mapping of sub-nanometre particles using energy-filtered transmission electron microscopy (EFTEM). Multivariate statistical analysis (MSA) generated reconstructed datasets where the signal from particles smaller than 1 nm in diameter was successfully isolated from the original noisy background. The technique has been applied to the characterization of oxide dispersion strengthened (ODS) reduced activation FeCr alloys, due to their relevance as structural materials for future fusion reactors. Results revealed that most nanometer-sized particles had a core–shell structure, with an Yttrium–Chromium–Oxygen-rich core and a nano-scaled Chromium–Oxygen-rich shell. This segregation to the nanoparticles caused a decrease of the Chromium dissolved in the matrix, compromising the corrosion resistance of the alloy.     

A replica technique for extracting precipitates from zirconium alloys for transmission electron microscopy analysis

A reliable two-stage carbon replica technique has been developed to extract precipitates from zirconium alloys. Using this technique, all precipitating phases can be extracted from Zircaloy-2, Zr-Cr-Fe, and Zr-Nb-Fe alloys. Precipitate identification using EDS X-ray analysis and convergent beam electron diffraction was greatly facilitated in comparison to thin foils. In addition, the sensitivity for the detection of trace elements in particles was increased using extraction replicas. The chemical compositions of the precipitates as determined from both replica and thin foils were in excellent agreement.     

Micellar calcium borate as an antiwear additive

The mechanisms of action of a new generation of antiwear additives is studied here by means of energy‐filtering transmission electron microscopy (EFTEM) carried out on the wear particles generated during friction tests between two ferrous surfaces (under boundary lubrication conditions). This paper deals with the structural and physico‐chemical changes that colloidal particles, calcium carbonate (CC) and calcium borate (CB) overbased salicylates detergents, have undergone during the build‐up of the interfacial antiwear tribofilm. EFTEM allowed us to investigate the nature of wear fragments originating from the film, stemming from CC and CB micelles, and to make a comparison regarding the tribofilm formation mechanisms. It appears that the CC wear debris are mainly crystalline and contain a high concentration of iron (as abrasive iron oxide Fe₂O₃), limiting their antiwear action. Consequently, CC micelles do not lead to an effective protective tribofilm. On the other hand, CB micelles do have an antiwear action, which we explained by the formation of a glassy iron borate tribofilm during the friction tests. Many of the CB wear fragments are composed of this amorphous material containing very small crystallites of residual calcite. Boron (contained in the CB micelles) is responsible for the presence of amorphous zones of the film and acts as a glass former, in a comparable way to phosphorus in zinc dithiophosphate. This revised version was published online in July 2006 with corrections to the Cover Date.     

Structural characterization of TiN/NbN multilayers: X-ray diffraction, energy-filtered TEM and Fresnel contrast techniques compared

Two TiN/NbN multilayers with wavelength 13.6 and 6.15 nm have been characterized by X-ray diffraction (XRD), Fresnel contrast analysis (FCA) and energy-filtered transmission electron microscopy (EFTEM). Good agreement between the composition profile obtained by FCA and EFTEM is obtained if the lower resolution of the EFTEM images is taken into account. The relative advantages and disadvantages of the techniques are discussed. Used together the two TEM techniques provide a quantitative characterization that is consistent with, and for some parameters provides more precise values than, that from XRD. The analysis shows that the multilayers have narrow interfaces (< 1 nm) and a composition amplitude close to 95% for the longer wavelength.     

Multivariate statistics applications in phase analysis of STEM-EDS spectrum images

Spectrum imaging (SI) methods are displacing traditional spot analyses as the predominant paradigm for spectroscopic analysis with electron beam instrumentation. The multivariate nature of SI provides clear advantages for qualitative analysis of multiphase specimens relative to traditional gray-scale images acquired with non-spectroscopic signals, where different phases with similar average atomic number may exhibit the same intensity. However, with the improvement in qualitative analysis with the SI paradigm has come a decline in the quantitative analysis of the phases thus identified, since the spectra from individual pixels typically have insufficient counting statistics for proper quantification. The present paper outlines a methodology for quantitative analysis within the spectral imaging paradigm, which is illustrated through X-ray energy-dispersive spectroscopy (EDS) of a multiphase (Pb,La)(Zr,Ti)O₃ ceramic in scanning transmission electron microscopy (STEM). Statistical analysis of STEM-EDS SI is shown to identify the number of distinct phases in the analyzed specimen and to provide better segmentation than the STEM high-angle annular dark-field (HAADF) signal. Representative spectra for the identified phases are extracted from the segmented images with and without exclusion of pixels that exhibit spectral contributions from multiple phases, and subsequently quantified using Cliff–Lorimer sensitivity factors. The phase compositions extracted with the method while excluding pixels from multiple phases are found to be in good agreement with those extracted from user-selected regions of interest, while providing improved confidence intervals. Without exclusion of multiphase pixels, the extracted composition is found to be in poor statistical agreement with the other results because of systematic errors arising from the cross-phase spectral contamination. The proposed method allows quantification to be performed in the presence of discontinuous phase distributions and overlapping phases, challenges that are typical of many nanoscale analyses performed by STEM-EDS.     

A facile method to compare EFTEM maps obtained from materials changing composition over time

Energy Filtered Transmission Electron Microscopy (EFTEM) is an analytical tool that has been successfully and widely employed in the last two decades for obtaining fast elemental maps in TEM mode. Several studies and efforts have been addressed to investigate limitations and advantages of such technique, as well as to improve the spatial resolution of compositional maps. Usually, EFTEM maps undergo post‐acquisition treatments by changing brightness and contrast levels, either via dedicated software or via human elaboration, in order to maximize their signal‐to‐noise ratio and render them as visible as possible. However, elemental maps forming a single set of EFTEM images are usually subjected to independent map‐by‐map image treatment. This post‐acquisition step becomes crucial when analyzing materials that change composition over time as a consequence of an external stimulus, because the map‐by‐map approach doesn't take into account how the chemical features of the imaged materials actually progress, in particular when the investigated elements exhibit very low signals. In this article, we present a facile procedure applicable to whole sets of EFTEM maps acquired on a sample that is evolving over time. The main aim is to find a common method to treat the images features, in order to make them as comparable as possible without affecting the information there contained. Microsc. Res. Tech. 78:1090–1097, 2015. © 2015 Wiley Periodicals, Inc.     

Energy-filtered transmission electron microscopy of biological samples on highly transparent carbon nanomembranes

Ultrathin carbon nanomembranes (CNM) comprising crosslinked biphenyl precursors have been tested as support films for energy-filtered transmission electron microscopy (EFTEM) of biological specimens. Due to their high transparency CNM are ideal substrates for electron energy loss spectroscopy (EELS) and electron spectroscopic imaging (ESI) of stained and unstained biological samples. Virtually background-free elemental maps of tobacco mosaic virus (TMV) and ferritin have been obtained from samples supported by ∼1 nm thin CNM. Furthermore, we have tested conductive carbon nanomembranes (cCNM) comprising nanocrystalline graphene, obtained by thermal treatment of CNM, as supports for cryoEM of ice-embedded biological samples. We imaged ice-embedded TMV on cCNM and compared the results with images of ice-embedded TMV on conventional carbon film (CC), thus analyzing the gain in contrast for TMV on cCNM in a quantitative manner. In addition we have developed a method for the preparation of vitrified specimens, suspended over the holes of a conventional holey carbon film, while backed by ultrathin cCNM.     

Preparation of location-specific thin foils from Fe–3% Si bi- and tri-crystals for examination in a FEG-STEM

Bi-crystals and tri-crystals of a nominal Fe–3% Si (wt%) of well-defined orientations have been grown using a floating-zone technique with optical heating. The manufacture of these unique crystals and the preparation technique involved in harvesting thin foils from specific locations for transmission electron microscopy are described in detail. In particular, the grain boundary triple junction has been extracted from the tri-crystal and examined in high-resolution aberration-corrected FEG-STEM instruments. To achieve the necessary resolution, the foils have to be uniformly thin, in the range 50–100 nm over large areas of the specimen. For ferromagnetic materials, there are further challenges arising from the magnetic field interaction, with the electron beam placing significant demands on the aberration correction system. One way to minimise this interaction is to reduce the total mass of magnetic material. To achieve this, an in situ focused ion beam lift-out technique has been combined with an additional precision ion-polishing stage to reproducibly provide thin-foil specimens suitable for high-resolution EELS and EDX analysis. Examination of the foils reveals that the final precision ion-polishing stage removes residual damage arising from the use of focused ion beam milling procedures.     

Spectroscopic electron tomography

The elemental mapping techniques in analytical transmission electron microscopy (TEM), energy filtered imaging (EFTEM) and EDX-mapping, are shown to provide new routes for tomographic reconstructions of 3D chemical maps on the nanoscale. The inelastic scattering does not only provide chemical sensitivity but also improves the linear projection relationship between mass density and image intensity, which often fails in bright field TEM of crystalline materials due to diffraction contrast. Instrumental requirements and artefact sources within the contrast formation mechanisms and within the numerical reconstruction are assessed.     

Quantification and thickness correction of EFTEM phosphorus maps

We describe a method for correcting plural inelastic scattering effects in elemental maps that are acquired in the energy filtering transmission electron microscope (EFTEM) using just two energy windows, one above and one below a core edge in the electron energy loss spectrum (EELS). The technique is demonstrated for mapping low concentrations of phosphorus in biological samples. First, the single-scattering EELS distributions are obtained from specimens of pure carbon and plastic embedding material. Then, spectra are calculated for different specimen thicknesses t, expressed in units of the inelastic mean free path lambda. In this way, standard curves are generated for the ratio k0 of post-edge to pre-edge intensities at the phosphorus L2,3 excitation energy, as a function of relative specimen thickness t/lambda. Thickness effects in a two-window phosphorus map are corrected by successive acquisition of zero-loss and unfiltered images, from which it is possible to determine a t/lambda image and hence a background k0-ratio image. Knowledge of the thickness-dependent k0-ratio at each pixel thus enables a more accurate determination of the phosphorus distribution in the specimen. Systematic and statistical errors are calculated as a function of specimen thickness, and elemental maps are quantified in terms of the number of phosphorus atoms per pixel. Further analysis of the k0-curve shows that the EFTEM can be used to obtain reliable two-window phosphorus maps from specimens that are considerably thicker than previously possible.     

Optimization of EFTEM image acquisition by using elastically filtered images for drift correction

Because of its high spatial resolution, energy-filtering transmission electron microscopy (EFTEM) has become widely used for the analysis of the chemical composition of nanostructures. To obtain the best spatial resolution, the precise correction of instrumental influences and the optimization of the data acquisition procedure are very important. In this publication, we discuss a modified image acquisition procedure that optimizes the acquisition process of the EFTEM images, especially for long exposure times and measurements that are affected by large spatial drift. To alleviate the blurring of the image caused by the spatial drift, we propose to take several EFTEM images with a shorter exposure time (sub-images) and merge these sub-images afterwards. To correct for the drift between these sub-images, elastically filtered images are acquired between two subsequent sub-images. These elastically filtered images are highly suitable for spatial drift correction based on the cross-correlation method. The use of the drift information between two elastically filtered images permits to merge the drift-corrected sub-images automatically and with high accuracy, resulting in sharper edges and an improved signal intensity in the final EFTEM image. Artefacts that are caused by prominent noise-peaks in the dark reference image have been suppressed by calculating the dark reference image from three images. Furthermore, using the information given by the elastically filtered images, it is possible to drift-correct a set of EFTEM images already during the acquisition. This simplifies the post-processing for elemental mapping and offers the possibility for active drift correction using the image shift function of the microscope, leading to an increased field of view.