Journal of Electron Microscopy - current issue

Development of an analytical environmental TEM system and its application

Sun, 15 Nov 2009 01:00:01 PST

Many automotive materials, such as catalysts and fuel cell materials, undergo significant changes in structure or properties when subjected to temperature change or the addition of a gas. For this reason, in the development of these materials, it is important to study the behavior of the material under controlled temperatures and gaseous atmospheres. Recently, a new environmental transmission electron microscope (TEM) has been developed for observation with a high resolution at high temperatures and under gaseous atmospheres, thus making it possible to analyze reaction processes in details. Also, the new TEM provides a high degree of reproducibility of observation conditions, thus making it possible to compare and validate observation of various specimens under a given set of conditions. Furthermore, easiness of gas condition and temperature control can provide a powerful tool for the studying of the mechanism of material change, such as oxidation and reduction reactions.

Correction of higher order geometrical aberration by triple 3-fold astigmatism field

Sun, 15 Nov 2009 01:00:02 PST

A new concept of a spherical aberration correction system using three dodecapoles is proposed. The system compensates for higher order aberration of 6-fold astigmatism, which generally limits a uniform phase area for image forming and probe forming in an electron microscope with a conventional two-hexapole corrector. Triple 3-fold astigmatism field is used to correct the spherical aberration of the objective lens, and the total 3-fold astigmatism is eliminated by their combination. The optimum azimuth relationship among three dodecapoles is calculated to eliminate the 6-fold astigmatism. The principle of the method was verified using a mathematically complex representation. This new concept was experimentally tested with a scanning transmission electron microscope at 60 kV acceleration. The 6-fold astigmatism was certainly compensated and the coherent convergent angle became almost twice compared to a conventional double hexapole system.

HREM and EDS analysis of (La, M)TiO3 (M = Zn, Mn) prepared by M/Li ion exchange

Tsurui, T., Wataru, M., Watanabe, M., Katsumata, T., Inaguma, Y. — Sun, 15 Nov 2009 01:00:02 PST

High-resolution electron microscopy (HREM) and energy dispersive X-ray spectroscopy (EDS) studies of (La, M)TiO₃ (M = Zn, Mn) prepared by M/Li ion exchange were performed to clarify the microscopic mechanism of ion-exchange reactions. At a macroscopic level, EDS analysis revealed that Zn and Mn atoms were homogeneously dispersed in matrix grains of (La, Zn)TiO₃ and (La, Mn)TiO₃ samples, respectively. EDS analysis also revealed that no segregation of Zn and Mn atoms was observed even in the vicinity of grain boundary regions. HREM and EDS analysis demonstrated that the microscopic mechanism of ion-exchange reactions was different by the ion-exchanged metals.

STEM imaging of 47-pm-separated atomic columns by a spherical aberration-corrected electron microscope with a 300-kV cold field emission gun

Sun, 15 Nov 2009 01:00:02 PST

A spherical aberration-corrected electron microscope has been developed recently, which is equipped with a 300-kV cold field emission gun and an objective lens of a small chromatic aberration coefficient. A dumbbell image of 47 pm spacing, corresponding to a pair of atomic columns of germanium aligned along the [114] direction, is resolved in high-angle annular dark field (HAADF) scanning transmission electron microscopy (STEM) with a 0.4-eV energy spread of the electron beam. The observed image was compared with a simulated image obtained by dynamical calculation.

Immunohistochemical localization of the STRO-1 antigen in developing rat teeth by light microscopy and electron microscopy

Kaneko, R., Akita, H., Shimauchi, H., Sasano, Y. — Sun, 15 Nov 2009 01:00:02 PST

STRO-1 is a cell-surface antigen. A cell population that is positive for the anti-STRO-1 antibody has been shown to contain mesenchymal stem cells. STRO-1-positive cells have been reported to reside in dental pulp and periodontal ligaments as well as in bone marrow. However, the tissue localization of STRO-1 in developing teeth is not clear. The present study was designed to investigate the spatiotemporal localization of STRO-1 in developing rat teeth by immunohistochemistry using light microscopy and electron microscopy. Wistar rats at ages 2, 3, 6 and 12 weeks postnatum and embryos at 18 days postcoitum were fixed with 4% paraformaldehyde. Mandibles and maxillae were resected and decalcified in 10% EDTA. The specimens were embedded in paraffin, and sections were cut and processed for immunohistochemistry using the anti-STRO-1 antibody. Selected specimens were frozen, sectioned and processed for immunoelectron microscopy. Immunoreactions for STRO-1 were identified in some bone marrow cells. Some odontoblasts and dental pulp cells showed positive immunoreactivity in developing rat tooth crowns and roots. Alveolar osteoblasts, cementoblasts and periodontal ligament cells were also immunoreactive. Electron microscopy localized the antigen in plasma membrane and some vesicles in dental pulp cells and odontoblasts. The study suggests that the STRO-1 antigen is involved in the differentiation of mesenchymal cell lineages and formation of the matrix in dental tissues.

Ultrastructural megakaryocyte modifications after vanadium inhalation in spleen and bone marrow

Sun, 15 Nov 2009 01:00:02 PST

Previous reports from our laboratory informed in mice an increase in platelets in blood, and megakaryocytes in spleen and bone marrow after vanadium inhalation. This element has become important in recent years because of its increased presence as an air pollutant. With this precedent, we evaluate the ultrastructural modifications in MKs from the spleen and bone marrow in our mouse experimental model. Mice inhaled 0.02 M V₂O₅ 1 h twice a week for 12 weeks. Tissues were processed for transmission electron microscopy. Results indicate an increase in the size and cytoplasmic granular content, as well as nuclear changes in MKs of exposed mice, changes which correlate with the time of exposure. Modifications in MKs described here suggest that inhaled vanadium induce megakaryocytic maturation, a raise in its granules content and demarcation membrane systems, which may lead to a rise in circulating platelet production and an increased risk for thromboembolic events.

FGF23 is mainly synthesized by osteocytes in the regularly distributed osteocytic lacunar canalicular system established after physiological bone remodeling

Sun, 15 Nov 2009 01:00:02 PST

This study aimed to evaluate whether the immunolocalization of fibroblast growth factor (FGF) 23 and dentin matrix protein 1 (DMP1) is associated with the spatial regularity of the osteocyte lacunar canalicular system(s) (OLCS). Femora of 12-weeks-old male ICR mice were fixed with 4% paraformaldehyde, decalcified with a 10% EDTA solution and then embedded in paraffin. We have devised a triple staining procedure that combines silver impregnation, alkaline phosphatase (ALPase) immunohistochemistry and tartrate-resistant acid phosphatase (TRAPase) enzyme histochemistry on a single paraffin section. This procedure permitted the visualization of ALPase-positive plump osteoblasts and several TRAPase-positive osteoclasts on those bone matrices featuring irregularly arranged OLCS, and of ALPase-positive bone lining cells on the bone matrix displaying the well-arranged OLCS. As observations proceeded from the metaphysis toward the diaphysis, the endosteal cortical bone displayed narrower bands of calcein labeling, accompanied by increased regularity of the OLCS. This implies that the speed of bone deposition during bone remodeling would affect the regularity of the OLCS. While DMP1 was evenly localized in all regions of the cortical bones, FGF23 was more abundantly localized in osteocytes of cortical bones with regularly arranged OLCS. In cortical bones, the endosteal area featuring regular OLCS exhibited more intense FGF23 immunoreaction when compared to the periosteal region, which tended to display irregular OLCS. In summary, FGF23 appears to be synthesized principally by osteocytes in the regularly distributed OLCS that have been established after bone remodeling.

Electron microscopy of octacalcium phosphate in the dental calculus

Kakei, M., Sakae, T., Yoshikawa, M. — Sun, 15 Nov 2009 01:00:02 PST

The purpose of this study was to morphologically demonstrate the presence of octacalcium phosphate in the dental calculus by judging from the crystal lattice image and its rapid transformation into apatite crystal, as part of our serial studies on biomineral products. We also aimed to confirm whether the physical properties of octacalcium phosphate are identical with those of the central dark lines observed in crystals of ordinary calcifying hard tissues. Electron micrographs showed that crystals of various sizes form in the dental calculus. The formation of each crystal seemed to be closely associated with the organic substance, possibly originating from degenerated microorganisms at the calcification front. Many crystals had an 8.2-Å lattice interval, similar to that of an apatite crystal. Furthermore, some crystals clearly revealed an 18.7-Å lattice interval and were vulnerable to electron bombardment. After electron beam exposure, this lattice interval was quickly altered to about half (i.e. 8.2 Å), indicating structural conversion. Consequently, a number of apatite crystals in the dental calculus are possibly created by a conversion mechanism involving an octacalcium phosphate intermediate. However, we also concluded that the calcification process in the dental calculus is not similar to that of ordinary calcifying hard tissues.