Experiments with an Imaging Filter in a CTEM

The resolution of a conventional transmission electron microscope (TEM) deteriorates as the specimen thickness increases, because chromatic aberration of the objective lens is caused by the energy loss of electrons). In the case of a scanning electron microscope (SEM), chromatic aberration does not exist as the restrictive factor for the resolution of the transmitted electron image, for the SEM has no imageforming lens. It is not sure, however, that the equal resolution to the probe diameter can be obtained in the case of a thick specimen. To study the relation between the specimen thickness and the resolution of the trans-mitted electron image obtained by the SEM, the following experiment was carried out.

IntroductionCurrently, the resolution of structural detail in conventional transmission electron microscopy (CTEM) of organic materials is limited by the electron beam damage suffered by the sample. It has recently been shown that the imaging modes now available in scanning transmission electron microscopy (STEM) provide the potential for improvements in resolution over CTEM methods. However, the effect of the sensitivity of an organic sample itself to beam damage has not been compared in STEM versus CTEM. In STEM each element of the sample area is briefly illuminated with a small diameter, high flux, scanning electron beam, whereas in CTEM the entire sample area is continuously illuminated with a large diameter, lower flux, static electron beam. Although it has been shown that the amount of sample damage in CTEM is independent of dose rate(2), the electron flux in STEM may range from 3 to 4 orders of magnitude greater than that in CTEM, with a possible influence on damage rate. Generally, the effects of beam size, high flux, and short dwell time of the scanning electron beam on sensitivity of the sample to damage in STEM are not known.

Applications of high voltage electron microscopy to botanical ultrastructure

The evolution of hydrogen from many hydrated cryo-preserved soft materials under electron irradiation in the transmission electron microscope can be observed at doses of the order of 1000 e nm(-2) and above. Such hydrogen causes artefacts in conventional transmission electron microscope or scanning transmission electron microscopy (STEM) imaging as well as in analyses by electron energy-loss spectroscopy. Here we show that the evolution of hydrogen depends on specimen thickness. Using wedge-shaped specimens of frozen-hydrated Nafion, a perfluorinated ionomer, saturated with the organic solvent DMMP together with both thin and thick sections of frozen-hydrated porcine skin, we show that there is a thickness below which hydrogen evolution is not detected either by bubble observation in transmission electron microscope image mode or by spectroscopic analysis in STEM electron energy-loss spectroscopy mode. We suggest that this effect is due to the diffusion of hydrogen, whose diffusivity remains significant even at liquid nitrogen temperature over the length scales and time scales relevant to transmission electron microscopy analysis of thin specimens. In short, we speculate that sufficient hydrogen can diffuse to the specimen surface in thin sections so that concentrations are too low for bubbling or for spectroscopic detection. Significantly, this finding indicates that higher electron doses can be used during the imaging of radiation-sensitive hydrated soft materials and, consequently, higher spatial resolution can be achieved, if sufficiently thin specimens are used in order to avoid the evolution of hydrogen-based artefacts.

The energy filtered imaging technique has so far been carried out in a scanning transmission electron microscope (STEM) fitted with a sector type energy analyzer. The STEM has a disadvantage of low beam parallelity because it uses a convergent beam, while the conventional transmission electron microscope (CTEM) allows good phase contrast and diffraction contrast to be obtained because of the high parallelity of the beam, and allows also high resolution images to be obtained. The technique to obtain energy filtered CTEM images has thus far been carried out by a Castaing-Henry type filter or an Ω type filter. However, these filters have the disadvantage of lower energy resolution than conventional sector type energy analyzer at the present time. This paper reports energy filtered CTEM images of MgO smoke, obtained using a new scanning CTEM image technique and a high energy resolution sector type energy analyzer which can resolve bulk and surface plasmon energy.

Adult channel catfish were exposed to 0 or 15 ppm arsenic as sodium arsenate in synthetic lake water for approximately six months. After sacrifice, livers were removed and examined by conventional transmission electron microscopy, optical microscopy, scanning electron microscopy, and electron probe X-ray microanalysis. Cytoplasmic structures in the nuclei were found to contain highly concentrated deposits of iron. Thirty percent of these structures were also found to contain arsenic at a level above the minimum detectable level for the conditions of the study. The size, shape and relative position of the cytoplasmic structures were the same as the hemosiderin granules observed by conventional transmission electron microscopy. (JMT)

6 Electron Spectroscopic Imaging: An Advanced Technique for Imaging and Analysis in Transmission Electron Microscopy

Whole-cell imaging of the budding yeast Saccharomyces cerevisiae by high-voltage scanning transmission electron tomography

Abstract: Contrast transfer function (CTF) is a vital function in transmission electron microscopy (TEM). It expresses to what extent amplitudes converted from the phase changes of the diffracted waves contribute to the TEM image, including the effects of lens aberrations. Simulation is very helpful to understand the application of the function thoroughly. In this work, we develop the CTFscope as a component in the Landyne software suite, to calculate the CTF with temporal and spatial dumping envelopes for conventional TEM and to extend it to various aberrations (up to fifth order) for aberration-corrected (AC)-TEM. It also includes effects on the CTF and imaging due to the objective aperture and image drift for tutorial purposes. The CTFscope has a user-friendly graphical interface for simulation, visualization, saving, and loading of the microscopy conditions and results. It can be used to explore instrumental performance information due to optical aberrations, effects on the resolution of a TEM and AC-TEM, and for teaching electron microscopy courses.

The effects of parenteral injection of aluminum, indium, gadolinium, or terbium in rats have been previously studied in several organs such as the liver, the kidneys, etc., but never in mammary glands. In this work, we have attempted to study the subcellular localization of these elements after their intraperitoneal administration. Their subsequent effects in the lactating mammary gland cells have also been studied. Our results using conventional transmission electron microscopy have shown that the lysosomes of the mammary glandular epithelial cells are the intracellular site of accumulation of the studied elements. Our results have also show intracellular deteriorations such as an expanded ergastoplasm and altered mitochondria after intraperitoneal injection of aluminum and indium.

The microstructural and morphological properties of the as-prepared tin (Sn) coating on ferrous substrates affects the lifetime as well as the applicability of the coating itself. In order to improve our knowledge of this system, strips of steel St-37 were coated with Sn in the form of gas-atomized powder, with an average diameter of 110±10 μm, using plasma spraying. The plasma gas was an Ar-He mixture and the current intensity was 650 A. The as-casted coating was characterized with scanning electron microscopy (SEM), X-ray diffraction (XRD) and conventional transmission electron microscopy (CTEM). It was deduced that the coating is characterized by high integrity, although certain pores were detected far from the Fe-Sn interface. However they refer to less than 1% of the total volume. Nevertheless the examination with SEM and XRD proved that the coating is mainly composed of pure Sn. However, interdiffusion takes place between the two phases (Fe-Sn) at their interface as deduced by CTEM. As a result, certain Fe-Sn phases are formed, which seem to be scattered in the coating close to the interface in the form of small inclusions.

High density lipoproteins of d = 1.12 to 1.21 g/ml from human serum (HDL3) were studied by electron microscopy with both negative staining and freeze-etching techniques. For the negatively stained specimens, a modified conventional transmission electron microscope as well as a scanning transmission electron microscope were used. The freeze-etched specimens were examined by a conventional transmission electron microscope. The diameter of HDL3 was found to be 105 +/- 4 A by freeze-etching and 94 +/- 6 A by negative staining. The surface of the HDL3 particles exhibited about 12 discrete domains, 28 +/- 3 A (freeze-etched) and 28 +/- 4 A (negatively stained) in diameter, of undefined chemical composition. Moreover, the freeze-etched specimens revealed an inner core 40 +/- 2 A in diameter, corresponding to estimated values reported previously. All information is consistent with the HDL3 model proposed by B. W. Shen, F. J. Kézdy, and A. M. Scanu [(1977) Proc. Natl. Acad. Sci. USA 74, 837-841], with additional evidence for well-defined surface substructure. The consistency of the images obtained with the various electron microscopy techniques and the marked change in the appearance of the surface in the HDL3 preparations that were digested by phospholipase A2 (EC 3.1.1.4) support the validity of the interpretation.

In many fields of research in science, engineering, and medicine, electron microscopy as a method for directly imaging submicroscopic structures has become increasingly important in recent decades. Electron microscopy (EM) includes several different techniques: conventional transmission electron microscopy (TEM), high-resolution electron microscopy (HREM), highvoltage electron microscopy (HVEM), scanning electron microscopy (SEM), analytical electron microscopy (AEM), emission electron microscopy (EEM), and others. In the past the central aim of using electron microscopy was structure determination, but recently it has been of growing importance for also investigating different processes, i.e., changes in materials by interaction with several influential factors (e.g., heat, electric or mechanic fields, mechanical loading). Of particular interest is the study of the micromechanical processes of deformation and fracture. Therefore, electron microscopy is a very powerful tool for materials science. The present review reports on some capabilities and limitation of the application of electron microscopy to solid polymers. In Section II the techniques of electron microscopy are briefly reviewed, followed by a section that describes the main methods of specimen preparation of solid polymeric materials. In Section IV the results of several applications of electron microscopy are discussed to reveal the morphology as well as the micromechanical deformation processes of several polymeric materials.

An amount of light elements contained in biological specimens is barely a few, so that the detection of them in the ultrathin sections is imposible. An analysis of electron energy loss (EEL) filter image is an ideal to detect such as phosphorus including in the sections. CEM-902 is a commercially available electron microscope to use for a such purpose. The authers attempted to get EEL filter images using the combined system of a conventional scanning transmission electron microscope (STEM) with EELS analyzer.The cultured cells with calcified substances and the liver, kidney, thyroid gland were used for biological specimens. There were fixed with 2.5% glutaraldehyde and with or without 1% osmium tetroxide, and then embedded in epoxy resin. 40 run thick sections were analyzed without any staining and carbon coating using a Carl Zeiss CEM-902 or a JEOL 2000EX with a EELS analyzer and a Tracor-Northern TN-5500, and with a Gatan cryo-transfer holder.

Correction of the Spherical Aberration of the Objective Lens in a Conventional Transmission Electron Microscope by Means of a Foil Lens