SummaryCellular structures of the fission yeast, Schizosaccharomyces pombe, were examined by using hard X-ray tomography. Since cells are nearly transparent to hard X-rays, Zernike phase contrast and heavy metal staining were introduced to improve image contrast. Through using such methods, images taken at 8 keV displayed sufficient contrast for observing cellular structures. The cell wall, the intracellular organelles and the entire structural organization of the whole cells were visualized in three-dimensional at a resolution better than 100 nm. Comparison between phase contrast and absorption contrast was also made, indicating the obvious advantage of phase contrast for cellular imaging at this energy. Our results demonstrate that hard X-ray tomography with Zernike phase contrast is suitable for cellular imaging. Its unique abilities make it have potential to become a useful tool for revealing structural information from cells, especially thick eukaryotic cells.

Light microscope observations suggest there are two discrete biochemical domains in the plant kinetochore, an inner domain containing structural proteins, and an outer domain containing proteins involved in motility. We analyzed the ultrastructure of maize meiotic kinetochores following high pressure freezing and freeze substitution, a method that provides excellent sample preservation. Data from meiosis II support previous descriptions of plant kinetochores as diffuse, nearly invisible domains, sometimes nesting in a cup of darkly staining chromatin. The ultrastructure is similar in meiosis I but there are two sister kinetochores that each protrude away from the chromosome and form their own distinct kinetochore fibers. Microtubules terminate within kinetochores where their ends are splayed in a cone-shaped configuration suggestive of microtubule disassembly. We could not detect any significant substructure within the kinetochore proper. We suggest that the diffuse structure classically defined as the kinetochore represents only the outer domain of a two-domain organelle. The inner domain, known to contain chromatin-binding proteins, probably extends into the electron-dense chromatin of the primary constriction.