The preparation of transmission electron microscope specimens from compound semiconductors by ion milling

It is shown that transmission electron microscope specimens prepared from compound semiconductors by conventional argon ion milling may contain serious artefactual strucutures. The nature of these artefacts is characterised and correlated in detail with specific features of the ion milling process. Results obtained using argon ion milling, for a number of semiconducting materials over a range of milling conditions, are compared with those obtained using alternative ion species. These comparative results demonstrate the great improvement in specimen structural quality which may be achieved by combining a logical selection of ion species with a careful choice of ion milling conditions. Full information is given on specific processes for producing thin specimens of high structural quality from a wide range of compound semiconductors.     

A transmission electron microscope study of the effects of ion etching on cells.

The effects of ion etching on blood cells have previously been studied by scanning electron microscopy. This present study by transmission electron microscopy was undertaken to evaluate the effects of the etching process on the cells. Critical point dried preparations were made, etched and subsequently processed and embedded in Araldite. Examination of thin sections of erythrocytes revealed disintegration of the plasma membrane; the residual membrane destruction products formed the tips of cones produced by long etching times. The effect of etching varied in erythrocytes in the same preparation. Nucleated cells showed a similar disintegration of the plasma membrane, but membranes of mitochondria, granules, vesicles and vacuoles did not exhibit effects of etching comparable to those of the plasma membranes. After treatment with a number of different fixatives, erythrocytes on carbon-coated copper grids were also etched and examined directly in a high voltage electron microscope at 1 MV. The effects were comparable to those seen in thin sections. To study the etch rates of biological materials, the resonant frequencies of quartz crystals were measured after application of thin films of albumen and cholesterol and again after these had been etched. the ratio of the frequency changes indicated that the etch rate of albumen was approximately 2-5 times that of cholesterol. The results are discussed in the light of theories of the mechanisms involved in ion etching.     

A simple liquid-He-cooled specimen stage for an ultra-high resolution transmission electron microscope

A simple top-entry specimen holder for an ultra-high resolution transmission electron microscope which has demonstrated 2.5 Å point-to-point resolution at 30 K is described. The stage has proved useful in studying low-temperature solid phases and phase transformations and is expected to be effective in reducing radiation damage for some organic specimens.     

Transmission electron microscope specimen preparation for exploring the buried interfaces in plan view

A relatively easy and convenient process for the preparation of transmission electron microscope specimens of buried interfaces is described. The method is based on the alignment and realignment of the specimen rotation centre during ion milling. The ion‐milling time interval in which good samples are obtained is substantially extended in this way.     

A specimen preparation technique for transmission electron microscopy of surface layers

A TEM specimen preparation method is described, with the aid of which electron transparent foils can be obtained across the external surface of a specimen. After careful pre-treatment, steel specimens have been electrolytically coated with nickel. Conventional thinning in a plane cutting the substrate-coating interface, gave thin foils displaying the internal structure as a function of depth under the initial free surface. The method has also been applied to minute metal particles, of dimensions too small to allow manipulating and foil preparation by conventional methods. Image examples are shown, and the applicability of the method is discussed.     

Special specimen preparation methods for image processing in transmission electron microscopy: a review.

One of the important developments in quantitative electron microscopy has been the application of optical and computer imaging methods to electron micrographs. In general these techniques of image analysis have been applied to electron micrographs from isolated biological structures prepared in the presence of various negative stains. To make full use of image processing techniques there are obvious advantages in preparing suitable specimens containing large areas of repeating features. However, the number of naturally occurring biological specimens exhibiting crystalline or paracrystalline features suitable for high resolution electron microscopy and subsequent image analysis is relatively small.Some recent experiments on the in vitro formation of crystalline and paracrystalline arrays from highly concentrated and purified isometric, filamentous and rod-like viruses is reviewed. The problems associated with the preparative procedures for producing two-dimensional and three-dimensional crystalline arrays are discussed together with the possibility of extending the negative staining-carbon film method for studying the gradual dissociation or assembly of viral components.     

A variation of transmission electron microscope sample preparation for VLSI analysis

Sample preparation for VLSI analysis is often slow due to long ion milling time and because the location of the thin area of the sample is difficult to control. By modifying the standard techniques used with a VCR Group (and perhaps other) mechanical dimpler, the ion milling time can be reduced to less than 30 min. and the location on the thinned area reasonably controlled. These modifications involve the use of a radiused edge on the dimpling tool, a rubber O-ring on the polishing tool, and not rotating the sample platen during polishing. The modifications to the dimpling and polishing tools allow more control of the geometry of the dimple, while not rotating the sample platen allows a thinner sample to be produced and permits the use of the sample translation micrometers to shift the location of the thinned area during polishing. The quality of samples produced using this modified procedure is equivalent to that obtained with the more standard methods.     

Tomography experiment of an integrated circuit specimen using 3 MeV electrons in the transmission electron microscope

The possibility of utilizing high-energy electron tomography to characterize the micron-scale three dimensional (3D) structures of integrated circuits has been demonstrated experimentally. First, electron transmission through a tilted SiO(2) film was measured with an ultrahigh-voltage electron microscope (ultra-HVEM) and analyzed from the point of view of elastic scattering of electrons, showing that linear attenuation of the logarithmic electron transmission still holds valid for effective specimen thicknesses up to 5 microm under 2 MV accelerating voltages. Electron tomography of a micron-order thick integrated circuit specimen including the Cu/via interconnect was then tried with 3 MeV electrons in the ultra-HVEM. Serial projection images of the specimen tilted at different angles over the range of +/-90 degrees were acquired, and 3D reconstruction was performed with the images by means of the IMOD software package. Consequently, the 3D structures of the Cu lines, via and void, were revealed by cross sections and surface rendering.     

Preparation of cross-sectional transmission electron microscopy samples by electron beam lithography and reactive ion etching

A cross-sectional sample preparation technique is described that relies on lithographic and dry-etching processing, thus avoiding metallographic polishing and ion milling. The method is capable of producing cross-sectional transmission electron microscopy samples with a large amount of transparent area (1 μm × 2.5 mm) which allows the examination of many patterned test sites on the same sample from the same chip of a silicon wafer. An example of the application of the technique is given for localized oxidation through a mask.     

Secondary electron detection in the scanning transmission electron microscope

In a dedicated scanning transmission electron microscope (STEM) secondary electron images with high spatial resolution and good contrast can be obtained. Two types of detector are described. These take into account the secondary electrons which depend on the post-specimen field strength of the objective lens. Due to the thinness of the samples and the collection geometry the images differ from those obtained in a convectional scanning microscope. Examples are given where secondary electron images augment the information obtained by the more commonly used imaging modes.     

Mechanical scanning of the specimen in the scanning electron microscope

A scanning electron microscope (SEM) system equipped with a motor drive specimen stage fully controlled with a personal computer (PC) has been utilized for obtaining ultralow magnification SEM images. This modem motor drive stage works as a mechanical scanning device. To produce ultra-low magnification SEM images, we use a successful combination of the mechanical scanning, electronic scanning, and digital image processing techniques. This new method is extremely labor and time saving for ultra-low magnification and wide-area observation. The option of ultra-low magnification observation (while maintaining the original SEM functions and performance) is important during a scanning electron microscopy session.     

A high-speed area detector for novel imaging techniques in a scanning transmission electron microscope

A scanning transmission electron microscope (STEM) produces a convergent beam electron diffraction pattern at each position of a raster scan with a focused electron beam, but recording this information poses major challenges for gathering and storing such large data sets in a timely manner and with sufficient dynamic range. To investigate the crystalline structure of materials, a 16×16 analog pixel array detector (PAD) is used to replace the traditional detectors and retain the diffraction information at every STEM raster position. The PAD, unlike a charge-coupled device (CCD) or photomultiplier tube (PMT), directly images 120–200 keV electrons with relatively little radiation damage, exhibits no afterglow and limits crosstalk between adjacent pixels. Traditional STEM imaging modes can still be performed by the PAD with a 1.1 kHz frame rate, which allows post-acquisition control over imaging conditions and enables novel imaging techniques based on the retained crystalline information. Techniques for rapid, semi-automatic crystal grain segmentation with sub-nanometer resolution are described using cross-correlation, sub-region integration, and other post-processing methods.     

A combined top and side entry specimen stage for a Hitachi H500 transmission electron microscope

The Hitachi H500 transmission electron microscope has been modified in order that both the top and side entry specimen stages may be fitted simultaneously. This made possible top entry multi-specimen operation up to a maximum magnification of × 100,000 and a resolution of 1.8 nm, combined with the normal side entry stage facilities of ±60° tilt, 0.45 nm resolution and × 400,000 magnification.     

A comparative study in the transmission electron microscope and scanning electron microscope of intracellular structures in sheep heart muscle cells

The internal cellular structures of the sheep ventricular myocardium have been comparatively studied in the transmission electron microscope (TEM) and in the scanning electron microscope (SEM). For TEM studies the tissue was prepared according to standard methods. Thick sections (10 μm) of paraffin embedded material were, after they had been deparaffinized in toluene, critical point dried, coated with gold and examined in the SEM. The comparative TEM and SEM investigations revealed very good correspondence, and it is evident that the described preparation procedure for SEM has preserved the fine structures of myofibrils, mitochondria, T-Tubules and sarcoplasmic reticulum in an excellent life-like pattern. Of special interest was the three-dimensional demonstration of triads and of circumferentially arranged T-tubules.     

Topographical contrast in the transmission electron microscope

An adaptation of the Foucault method for topographical imaging in the transmission electron microscope is described in detail. The image contrast is produced by selection of electrons which have suffered differential phase retardations in the specimen inner potential. Surface or interface displacements produce bright or dark image contrast, and the ultimate resolution approaches that of the atomic scale. The imaging method is applied in studies of both amorphous and crystalline objects. The possibility of performing quantitative measurements is demonstrated by the estimation of the inner potential of crystalline MgO.     

Specimen preparation methods for the examination of surfaces and interfaces in the transmission electron microscope

Various techniques for the preparation of cross-sectional and plan view TEM specimens of surfaces and interfaces are described. Particular emphasis is given to preparative methods which are both generally applicable and which minimize differential thinning of the materials present on either side of the interface of interest, thereby improving the reliability of the approach.     

Development of a specimen holder for in situ generation of pure in-plane magnetic fields in a transmission electron microscope

As the development of quantised storage media progresses, detailed knowledge is required about the magnetisation reversal behaviour of sub-micron sized magnetic structures in external magnetic fields. Using the Lorentz mode of transmission electron microscopy (LTEM) the magnetic microstructure of thin film samples can be imaged with high spatial resolution. A novel approach for in situ magnetising experiments is described which combines the development of a custom-made sample holder which generates two orthogonal in-plane components of magnetic field in the specimen plane with the benefit of computer-controlled variation of the field. We present a specimen stage suitable for a Philips CM30 Twin/LTEM, which allows the generation of well-defined magnetic in-plane fields in the TEM.