Taiga Okumura

Characteristics of biogenic calcite in the prismatic layer of a pearl oyster, Pinctada fucata

The fine structure of the calcite prism in the outer layer of a pearl oyster, Pinctada fucata, has been investigated using various electron beam techniques, in order to understand its characteristics and growth mechanism including the role of intracrystalline organic substances. As the calcite prismatic layer grows thicker, sinuous boundaries develop to divide the prism into a number of domains. The crystal misorientation between the adjacent domains is several to more than ten degrees. The component of the misorientation is mainly the rotation about the c-axis. There is no continuous organic membrane at the boundaries. Furthermore, the crystal orientation inside the domains changes gradually, as indicated by the electron back-scattered diffraction (EBSD) in a scanning electron microscope (SEM). Transmission electron microscopy (TEM) examination revealed that the domain consists of sub-grains of a few hundred nanometers divided by small-angle grain boundaries, which are probably the origin of the gradual change of the crystal orientation inside the domains. Spherular Fresnel contrasts were often observed at the small-angle grain boundaries, in defocused TEM images. Electron energy-loss spectroscopy (EELS) indicated the spherules are organic macromolecules, suggesting that incorporation of organic macromolecules during the crystal growth forms the sub-grain structure of the calcite prism.

Direct observation of cesium at the interlayer region in phlogopite mica

To investigate the sorption mechanism of cesium (Cs) into clay minerals, high-resolution (scanning) transmission electron microscopy (TEM/STEM) imaging of Cs in mica (phlogopite) has been conducted. Platy phlogopite powders were immersed in a cesium chloride (CsCl) solution to achieve Cs(+)-K(+) ion-exchange at the interlayer regions in phlogopite. To observe many phlogopite particles with the incident electron beam parallel to the mica layers, cross-sectional thin specimens were prepared from sedimented particles using a focused ion beam. High-angle annular dark-field imaging with STEM is superior to conventional high-resolution TEM (HRTEM) for visualizing Cs at interlayer sites even in thicker crystal regions and/or at lower magnification due to the intense Z-contrast of Cs. However, HRTEM is also practical for estimating the concentration of Cs at the interlayer site from the thickness dependence of the contrast at the interlayer region. Cs sorption of micas was previously thought to be localized mainly at the frayed-edge sites of mica crystals. However, the present observations indicate that Cs substitution of K occurs not around crystal edges but deep inside the crystals along specific interlayer regions.

Loss of radioactivity in radiocesium-bearing microparticles emitted from the Fukushima Dai-ichi nuclear power plant by heating

Radiocesium-bearing microparticles (CsPs) substantially made of silicate glass are a novel form of radiocesium emitted from the broken containment vessel of Fukushima Dai-ichi nuclear power plant. CsPs have a potential risk of internal radiation exposure caused by inhalation. Radiation-contaminated waste including CsPs is being burned in incinerators; therefore, this study has investigated the responses of CsPs to heating in air. The radioactivity of CsPs gradually decreased from 600 °C and was almost lost when the temperature reached 1000 °C. The size and spherical morphology of CsPs were almost unchanged after heating, but cesium including radiocesium, potassium and chlorine were lost, probably diffused away from the CsPs. Iron, zinc and tin originally dissolved in the glass matrix were crystallized to oxide nanoparticles inside the CsPs. When the CsPs were heated together with weathered granitic soil that is common in Fukushima, the radiocesium released from CsPs was sorbed by the surrounding soil. From these results, it is expected that the radioactivity of CsPs will be lost when radiation-contaminated waste including CsPs is burned in incinerators.

Chitin Degraded by Chitinolytic Enzymes Induces Crystal Defects of Calcites

Mollusk shells have unique microstructures and mechanical properties such as hardness and flexibility. Calcite in the prismatic layer of P. fucata is extremely tough due to small crystal defects and localized organic networks inside calcites. Electron microscopic observations have suggested that such crystal defects are caused by the organic networks during calcite formation. Our previous work reported that the chitin which is the main component of organic networks and chitinolytic enzymes that bind to chitin were identified. In this article, to investigate the effects of chitin and chitinolytic enzymes on the formation of calcites, calcites were synthesized in chitin gel after treatment with chitinolytic enzymes. Chitin fibers seemed to become smooth and loosened after degradation. The crystal defects became larger as the chitin fibers became more degraded by chitinolytic enzymes in a dose-dependent manner. These results suggest that the shape of chitin fiber, which is regulated by the degradation of chitinolytic enzymes, contributes to the formation of small crystal defects.

On the Transition Temperature to Calcite and Cell Lengths for Various Biogenic Aragonites

In order to understand the mineralogical difference between biogenic aragonites and their geological or synthetic ones, the transition temperature from aragonite to calcite by heating and the cell lengths of a number of biogenic aragonites have been measured using conventional and high-temperature XRD, as well as those of abiotic ones. Among 21 specimens, most biogenic aragonites showed a transition temperature 60–100 °C lower than that for abiotic ones. However, the shells of land snails showed almost similar transition temperatures. The temperature range from the beginning to the completion of the transition was also varied among the biogenic aragonites. On the other hand, the axial ratios (a/b and c/b) of aragonites in marine molluscan species were considerably larger than those of abiotic ones. However, aragonites in freshwater molluscan species and land snails showed axial ratios similar to abiotic ones. X-ray microanalysis suggested that the origin of such abnormal cell lengths was sodium incorporated in the aragonite crystals, not due to lattice distortion induced by the intracrystalline organic molecules proposed in previous researches.

Electron tomography of whole cultured cells using novel transmission electron imaging technique

Since a three-dimensional (3D) cellular ultrastructure is significant for biological functions, it has been investigated using various electron microscopic techniques. Although transmission electron microscopy (TEM)-based techniques are traditionally used, cells must be embedded in resin and sliced into ultrathin sections in sample preparation processes. Block-face observation using a scanning electron microscope (SEM) has also been recently applied to 3D observation of cellular components, but this is a destructive inspection and does not allow re-examination. Therefore, we developed electron tomography using a transmission electron imaging technique called Plate-TEM. With Plate-TEM, the cells cultured directly on a scintillator plate are inserted into a conventional SEM equipped with a Plate-TEM observation system, and their internal structures are observed by detecting scintillation light produced by electrons passing through the cells. This technology has the following four advantages. First, the cells cultured on the plate can be observed at electron-microscopic resolution since they remain on the plate. Second, both surface and internal information can be obtained simultaneously by using electron- and photo-detectors, respectively, because a Plate-TEM detector is installed in an SEM. Third, the cells on the scintillator plate can also be inspected using light microscopy because the plate has transparent features. Finally, correlative observation with other techniques, such as conventional TEM, is possible after Plate-TEM observation because Plate-TEM is a non-destructive analysis technique. We also designed a sample stage to tilt the samples for tomography with Plate-TEM, by which 3D organization of cellular structures can be visualized as a whole cell. In the present study, Mm2T cells were investigated using our tomography system, resulting in 3D visualization of cell organelles such as mitochondria, lipid droplets, and microvilli. Correlative observations with various imaging techniques were also conducted by successive observations with light microscopy, SEM, Plate-TEM, and conventional TEM. Consequently, the Plate-TEM tomography technique encourages understanding of cellular structures at high resolution, which can contribute to cellular biological research.

Secondary radiocesium contamination of agricultural products by resuspended matter

Contamination of agricultural products by resuspended matter remains a concern in the highly contaminated areas. Radiocesium concentration of spinach cultivated with non-contaminated soil was low in the decontaminated areas, but high in the contaminated areas. The washed plants had relatively lower radiocesium concentration than the unwashed plants. Furthermore, the plants cultivated closer to the ground surface tended to have a higher radiocesium concentration than those cultivated farther from the ground. Therefore, it can be concluded that radiocesium found in the spinach leaves derived from resuspended matter in the air. With further analysis, radiocesium in the resuspended matter was confirmed to be present as particles.

Whole-Cell Tomography Using a Conventional Scanning Electron Microscope

Traditionally, ultrastructure of biological samples has been elucidated predominantly using transmission electron microscopy (TEM). For TEM observation, the samples are embedded in resin and cut into ultrathin sections using an ultramicrotome. These sample preparation procedures, however, require skills to make adequate electron-transparent thin sections on a small grid, which is obstacle to TEM examination. To overcome the difficulty, we developed plate-transmission electron microscopy (PlateTEM), by which internal structure of biological samples is observed using a transmitted electron (TE) mode of scanning electron microscopy (SEM) with a scintillator plate [1,2]. Furthermore, whole-cell tomography was conducted using this technique in order to image the three-dimensional (3D) structure of the samples.