Lorentz Scanning Electron/Ion Microscopy

We have developed an observation and measurement method for spatial electromagnetic fields by using scanning electron/ion microscopes, combined with electron holography reconstruction technique. A cross-grating was installed below the specimen, and the specimens were observed under the infocus condition, and the grating was simultaneously observed under the defocus condition. Electromagnetic fields around the specimen were estimated from grating-image distortions. This method is effective for low and middle magnification and resolution ranges; furthermore, this method can in principle be realizable in any electron/ion beam instruments because it is based on the Lorentz force model for charged particle beams. Mini Abstract We have developed a visualization technique for spatial electromagnetic fields by using scanning electron/ion microscopes, combined with electron holography reconstruction technique. A specimen and a cross-grating installed below the specimen were observed simultaneously. The distorted grating image caused by electromagnetic fields around the specimen were quantitatively measured and visualized.

Distribution of amniotic stem cells in human term amnion membrane

Amnion membrane studies related to miscarriage have been conducted in the field of obstetrics and gynecology. However, the distribution of stem cells within the amnion and the differences in the properties of each type of stem cells are still not well understood. We address this gap in knowledge in the present study where we morphologically classified the amnion membrane, and we clarified the distribution of stem cells here to identify functionally different amniotic membrane–derived stem cells. The amnion can be divided into a site that is continuous with the umbilical cord (region A), a site that adheres to the placenta (region B), and a site that is located opposite the placenta (region C). We found that human amnion epithelial stem cells (HAECs) that strongly express stem cell markers were abundant in area A. HAEC not only expressesed stem cell-specific surface markers TRA-1-60, Tra-1-81, SSEA4, SSEA3, but was also OCT-3/4 positive and had alkaline phosphatase activity. Human amniotic mesenchymal stem cells expressed KLF-A, OCTA, Oct3/4, c-MYC and Sox2 which is transcription factor. Especially, in regions A and B they have expressed CD73, and the higher expression of BCRP which is drug excretion transporter protein than the other parts. These data suggest that different types of stem cells may have existed in different area. The understanding the relation with characteristics of the stem cells in each area and function would allow for the efficient harvest of suitable HAE and HAM stem cells as using tool for regenerative medicine.

Cryo-Electron Microscopy of the Giant Viruses

High resolution study of the giant viruses presents one of the latest challenges in cryo-electron microscopy of viruses. Too small for light microscopy, but too large for easy study at high resolution by electron microscopy, they range in size from ~0.2-2 μm, from high symmetry icosahedral viruses such as Paramecium burseria Chlorella virus 1 to asymmetric forms like Tupanvirus or Pithovirus. To attain high resolution, two strategies exist to study these large viruses by cryo-EM: firstly, increasing the acceleration voltage of the electron microscope to improve sample penetration and overcome the limitations imposed by electro-optical physics at lower voltages, and secondly the method of “block-based reconstruction” pioneered by Michael G. Rossmann and his collaborators, which resolves the latter limitation through an elegant leveraging of high symmetry, but cannot overcome sample penetration limitations. In addition, more recent advances in both computational capacity and image processing also yield assistance in studying the giant viruses. Especially, the inclusion of Ewald sphere correction can provide large improvements in attainable resolutions for 300 kV electron microscopes. Despite this, the study of giant viruses remains a significant challenge.