title: A method for observing silver-stained osteocytes in situ in 3-microm sections using ultra-high voltage electron microscopy tomography.
abstract: Osteocytes are surrounded by hard bone matrix, and it has not been possible previously to directly observe the in situ architecture of osteocyte morphology in bone. Electron microscope tomography, however, is a technique that has the unique potential to provide three-dimensional (3D) visualization of cellular ultrastructure. This approach is based on reconstruction of 3D volumes from a tilt series of electron micrographs of cells, and resolution at the nanometer level has been achieved. We applied electron microscope tomography to thick sections of silver-stained osteocytes in bone using a Hitachi H-3000 ultra-high voltage electron microscope equipped with a 360 degrees tilt specimen holder, at an accelerating voltage of 2 MeV. Osteocytes with numerous processes and branches were clearly seen in the serial tilt series acquired from 3-microm-thick sections. Reconstruction of young osteocytes showed the 3D topographic morphology of the cell body and processes at high resolution. This morphological data on osteocytes should provide useful information to those who study osteocyte physiology and the several models used to explain their mechanosensory properties.
authors.affiliation: Department of Orthodontics, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama, Japan. kamioka@md.okayama-u.ac.jp
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Kamioka
authors.forename: Sakhr A
authors.initials: SA
authors.lastname: Murshid
authors.forename: Yoshihito
authors.initials: Y
authors.lastname: Ishihara
authors.forename: Naoko
authors.initials: N
authors.lastname: Kajimura
authors.forename: Toshiaki
authors.initials: T
authors.lastname: Hasegawa
authors.forename: Ryoko
authors.initials: R
authors.lastname: Ando
authors.forename: Yasuyo
authors.initials: Y
authors.lastname: Sugawara
authors.forename: Takashi
authors.initials: T
authors.lastname: Yamashiro
authors.forename: Akio
authors.initials: A
authors.lastname: Takaoka
authors.forename: Teruko
authors.initials: T
authors.lastname: Takano-Yamamoto
mesh_terms: D000818:Animals; D002645:Chickens; D004396:Coloring Agents / Q000494:pharmacology*; D055032:Electron Microscope Tomography / Q000379:methods*; D021621:Imaging, Three-Dimensional; D010011:Osteocytes / Q000648:ultrastructure*; D012834:Silver / Q000494:pharmacology*; D012886:Skull / Q000648:ultrastructure*; D013194:Staining and Labeling / Q000379:methods*
chemical_list: D004396:Coloring Agents; D012834:Silver