Imaging and spectrometry of a microscopic gaseous target by SEM and TEM

Microscopic gas jets are experimentally investigated using a scanning electron microscope/electron microprobe (JXA 50A, JEOL, Tokyo) equipped with a transmitted electron detector, and the CEM 902 (Zeiss, Oberkochen) which allows electron spectroscopic imaging. Both capillary microjet devices and circular orifices have been used. Imaging of the microgas jet (argon, air, artificial gas mixtures) is done by transmitted electrons (dark field), electrons as collected by the conventional Everhart-Thomley secondary electron detector, and by electron spectroscopic imaging, both at the argon M- and L edges. X-ray spectra and electron energy loss spectra taken at an axis point just a few micrometers downstream from the orifice are discussed.     

Sequential treatment by phosphotungstic acid and uranyl acetate enhances the adherence of lipid membranes and membrane proteins to hydrophobic EM grids

A simple procedure for screening by electron microscopic observations of conditions for the reconstitution of membrane proteins into lipid bilayers is described. This procedure consists of a 5–10-s treatment of electron microscopic grids, to which the sample has already been applied, with 1% phosphotungstic acid before proceeding with final staining in uranyl acetate. The method substantially enhances the adherence of lipid membranes and membrane protein particles to hydrophobic collodion/carbon grids.     

Measurement of specimen thickness by phase change determination in TEM

A non-destructive method for measuring the thickness of thin amorphous films composed of light elements has been developed. The method employs the statistics of the phase of the electron exit wave function. The accuracy of this method has been checked numerically by the multislice method and compared with that based on the mean inner potential.     

Observation of dislocations and microplasma sites in semiconductors by direct correlations of STEBIC,STEM and ELS

A high voltage electron microscope, equipped with scanning transmission (STEM) attachment, electron beam induced conductivity (EBIC) facilities, and electron energy loss spectrometer (ELS), has been used to investigate semiconductor devices. The capability of STEM to produce, simultaneously or sequentially, conductive and transmission images of the same specimen region, which can also be ELS analysed, is exploited in order to establish direct and unambiguous correlations between EBIC and STEM images of defective regions (dislocations and microplasma sites) in silicon devices. The results obtained are discussed in terms of correlations, resolution, contrast, and radiation damage; in addition, a comparison is made between this method and the other correlation methods based on EBIC/SEM (scanning electron microscope) and TEM (transmission electron microscope).     

Characterization of an oil-degrading Microcoleus consortium by means of confocal scanning microscopy, scanning electron microscopy and transmission electron microscopy

A consortium of microorganisms with the capacity to degrade crude oil has been characterized by means of confocal laser scanning microscopy (CLSM), transmission electron microscopy (TEM), and scanning electron microscopy (SEM). The analysis using CLSM shows that Microcoleus chthonoplastes is the dominant organism in the consortium. This cyanobacterium forms long filaments that group together in bundles inside a mucopolysaccharide sheath. Scanning electron microscopy and transmission electron microscopy have allowed us to demonstrate that this cyanobacterium forms a consortium primarily with three morphotypes of the heterotrophic microorganisms found in the Microcoleus chthonoplastes sheath. The optimal growth of Microcoleus consortium was obtained in presence of light and crude oil, and under anaerobic conditions. When grown in agar plate, only one type of colony (green and filamentous) was observed.     

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.     

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.     

Characterization of the annealed (0001) surface of sapphire (alpha-Al2O3) and interaction with silver by reflection electron microscopy and scanning reflection electron microscopy.

Annealed (0001) surfaces of single-crystal sapphire (alpha-Al2O3) rod have been studied in the electron microscope using reflection electron microscopy (REM), scanning reflection electron microscopy (SREM), and reflection high energy electron diffraction (RHEED). Annealed surfaces of (0001) sapphire are vicinal and characterized by close-packed (0001)-oriented terraces separated by faceted multiple-height steps, with edges parallel to energetically preferred low-index directions (less than 1010 greater than and less than 1120 greater than). These structural features are not seen on cleaved surfaces or polished surfaces treated at temperatures less than 1,250 degrees C. Oxygen-annealing produces clean surfaces which prove useful for investigating the interaction of deposited metals with the (0001) sapphire. Both REM and SREM (with microdiffraction spots) techniques have been used to observe fine structure of flat Ag islands on the scale of 1-100 nm on the (0001)-oriented terraces as well as aggregations at the steps. A preliminary result on interaction with Cu is also included.     

An ultrahigh vacuum system for in situ studies of thin films and nanostructures by nuclear resonance scattering of synchrotron radiation

A multifunctional ultrahigh vacuum (UHV) system has been set up at the nuclear resonance beamline ID18 of the European Synchrotron Radiation Facility (ESRF). Thin and ultrathin films, nanoislands and -wires, multilayers, and stoichiometric oxides can be prepared by molecular beam epitaxy and characterized by low-energy electron diffraction, Auger electron spectroscopy, and reflection high-energy electron diffraction. Upon characterization the sample is transferred under UHV conditions to the chamber for experiments with the synchrotron beam. Electronic and magnetic properties, vibrational dynamics, and diffusion phenomena can be investigated by several synchrotron radiation based techniques, such as nuclear forward scattering, nuclear inelastic and quasielastic scattering, synchrotron radiation based perturbed angular correlations, and nuclear and electronic reflectivity. In addition, two portable UHV chambers serve to transfer the sample to other beamlines profiting from the available experimental techniques at the ESRF.     

Reduction of polymer growth in electron microscopes by use of a fluorocarbon oxide pump fluid

A commonly experienced contamination problem in electron microscopes is the growth of electron induced polymer films on the electrodes and specimens in instruments evacuated by oil pumps. Hydrocarbon fluids are generally used in the vacuum pumps and experiments have been made to reduce the contamination rate in a transmission instrument (AEI 802) by using fractional distillates of a new fluorocarbon oxide compound. The rotary pump was replaced by one charged with a fluorocarbon oxide lubricant and the fluid charge in the diffusion pump replaced by a fluorocarbon oxide evaporant. Both specimen and electrode contamination were greatly reduced by the use of the new fluid. Electron micrographs of a gold film are shown comparing contamination levels produced in organic and fluorocarbon oxide atmospheres after prolonged exposure to the electron beam.     

Characterization of polymer blends and block copolymers by conventional and low voltage SEM

Methods for characterizing block copolymers and polymer blends have been developed. Results from unstained and ruthenium tetroxide-stained samples obtained by scanning electron microscopy at various acceleration voltage and by transmission electron microscopy are presented. The contrast in secondary images between components in stained polymer blends, where one component is preferentially stained, is maximized at higher acceleration voltage (10–25 keV). For measurement of particle size and shape, this is the preferred operating condition. To obtain high-resolution images showing surface topography and fine structure, such as 20 nm domains in block copolymers, low-voltage (<5 keV) imaging is preferred. Observation of the 20 nm domain structure in hydrogenated styrenebutadiene-styrene shows that the spatial resolution now possible by scanning electron microscopy is comparable to that obtained by the traditional method of transmission electron microscopy.     

Electron-beam-induced reduction of Fe in iron phosphate dihydrate,ferrihydrite, haemosiderin and ferritin as revealed by electron energy-loss spectroscopy

The effect of high-energy electron irradiation on ferritin/haemosiderin cores (in an iron-overloaded human liver biopsy), its mineral analogue; six-line ferrihydrite (6LFh), and iron phosphate dihydrate (which has similar octahedral ferric iron to oxygen coordination to that in ferrihydrite and ferritin/haemosiderin cores) has been investigated using electron energy-loss spectroscopy (EELS). Fe L_2,3-ionisation edges were recorded on two types of electron microscope: a 200 keV transmission electron microscope (TEM) and a 100 keV scanning transmission electron microscope (STEM), in order to investigate the damage mechanisms in operation and to establish a methodology for minimum specimen alteration during analytical electron microscopic characterisation. A specimen damage mechanism dominated by radiolysis that results in the preferential loss of iron co-ordinating ligands (O, OH and H₂O) is discussed. The net result of irradiation is structural re-organisation and reduction of iron within the iron hydroxides. At sufficiently low electron fluence and particularly in the lower incident energy, finer probe diameter STEM, the alteration is shown to be minimal. All the materials examined exhibit damage which as a function of cumulative fluence is best fitted by an inverse power-law, implying that several chemical and structural changes occur in response to the electron beam and we suggest that these are governed by secondary processes arising from the primary ionisation event. This work affirms that electron fluence and current density should be considered when measuring mixed valence ratios with EELS.     

A method for the characterization of surface-modified asbestos fibres by electron energy-loss spectroscopy and electron spectroscopic imaging

A method for the characterization of surface-treated asbestos fibres with electron microscopy is presented. Electron spectroscopic imaging (ESI) of organosilane-treated chrysotile asbestos fibres has been carried out. Initially, the region below the carbon edge was inspected in ESI mode for its effectiveness as a background correction. Elemental mapping was performed on standard untreated fibres to take into account non-characteristic signals from extrapolation errors and camera artefacts. The highest resulting pixel value that results from non-characteristic signals was used as a threshold for further background correction in the net images. Samples for electron energy-loss spectroscopy were prepared in two different ways, either by gluing on grids, or by using perforated carbon foils. The results show that the use of a conducting carbon film is necessary for the analysis of such electrically insulating asbestos fibres. Focusing of the electron beam on the individual fibres results in a thermal effect promoting the evaporation of the organosilane reaction products.     

Automatic selection of macromolecules from electron micrographs by component labelling and symbolic processing

A new solution to the problem of extracting images of individual biological macromolecules from electron micrographs is described. There are three distinct steps in the process. The initial stage of low-level image processing consists of noise suppression and edge detection. An intermediate stage of component labelling and feature computation bridges the gap between the iconic (low-level) processing and the final phase of symbolic (high-level) processing. Simple symbolic objects (bounding boxes) are derived from the edges, and are easily represented and manipulated in the decision-making process. The efficacy of the algorithm is demonstrated using electron micrographs of ribosomes and ribosomal subunits. The hierarchical nature of the analysis embodies a reduction in the amount of data and a change in its nature. Initially, thousands of pixels of continuous gray levels must be dealt with. After component labelling, there are fewer than a hundred bounding boxes whose manipulation can easily be defined and articulated by an expert. The software package that has been written can thus serve as a basis for applying artificial intelligence methodologies to analysis of electron micrographs.     

Statistical significance of molecule projections by single particle averaging

A statistical test based on a multiple comparison of Friedman's rank sums has been developed to determine the statistical significance of molecule projections obtained by non-crystallographic averaging techniques. The test, applied to electron microscope images of the acetylcholine receptor molecule, confirms the significance and quoted resolution of averages by Zingsheim et al. (1980, 1982a, b).     

Parameters of the Electron Beams Generated by the РАДАН-220 and РА ДАН-ЭКСПЕРТ Accelerators

The measured parameters of the electron beams generated by small-sized nanosecond direct-action Р АДАН-220 and РАДАН-ЭКСП Е РТ accelerators are presented. The measuring techniques and the designs of electron detectors developed for this purpose are described. The fluence of electrons and the energy density, uniformity, and energy spectrum of the electron beam have been measured.     

Time resolved measurements by the pulse height analysis soft x-ray diagnostic on TCV

A single chord, single processing chain, hybrid (analog/digital) pulse height analysis diagnostic has been developed for the TCV tokamak, aiming to provide the evolution of the plasma electron temperature with a software selectable minimum temporal resolution of 100 ms. The high count rate (approximately 65 kHz) together with an energy resolution of 190 eV (at 5.9 keV) were achieved by encoding the data stream with an on-site developed interface amplifier and time generator. The diagnostic was also used to investigate the non-Maxwellian behavior of the electron energy distribution function with strong electron cyclotron resonance heating and to monitor the presence of intrinsic and injected impurities in the 700 eV-20 keV energy range. The conversion of this diagnostic into a real-time control tool is under development.     

7050铝合金的表面增压喷丸纳米化

在普通喷丸设备上加增压罐,采用增压喷丸方法对7050铝合金表面进行喷丸处理;用X射线衍射仪、光学显微镜、透射电镜、选区电子衍射仪等对合金表层结构进行了观察与分析,并用显微硬度计测试了表层的硬度。结果表明:经增压喷丸处理后7050铝合金表层的衍射峰明显宽化,随着处理时间的增加,塑性变形层厚度增加,表层晶粒细化至纳米级,平均晶粒尺寸约为40 nm;纳米化后的合金表层硬度比心部硬度提高了1.5倍,其原因可归结为细晶强化及加工硬化。     

Determination of gold-labelled surface receptors on single cells by X-ray microanalysis

A standardless X-ray microanalytical procedure has been developed to determine the number of gold-labelled surface receptors on whole single cells. The effect of the injection of K₂PtCl₄ into mice on gold-labelled concanavalin A (Con A) receptors on peritoneal macrophages was examined with an energy dispersive X-ray detector in an SEM. The numbers of gold particles seen in electron micrographs and estimated by fluorescence photometric measurements of fluorescein isothiocyanate-labelled Con A receptors were correlated with the X-ray microanalytical results.     

Morphology visualization of irregular shape bacteria by electron holography and tomography

In the current work, irregular morphology of Staphylococcus aureus bacteria has been visualized by phase retrieval employing off‐axis electron holography (EH) and 3D reconstruction electron tomography using high‐angle annular dark field scanning transmission electron microscopy (HAADF‐STEM). Bacteria interacting with gold nanoparticles (AuNP) acquired a shrunken or irregular shape due to air dehydration processing. STEM imaging shows the attachment of AuNP on the surface of cells and suggests an irregular 3D morphology of the specimen. The phase reconstruction demonstrates that off‐axis electron holography can reveal with a single hologram the morphology of the specimen and the distribution of the functionalized AuNPs. In addition, EH reduces significantly the acquisition time and the cumulative radiation damage (in three orders of magnitude) over biological samples in comparison with multiple tilted electron expositions intrinsic to electron tomography, as well as the processing time and the reconstruction artifacts that may arise during tomogram reconstruction.