A method for observing silver-stained osteocytes in situ in 3-microm sections using ultra-high voltage electron microscopy tomography.

Hiroshi Kamioka, Sakhr A. Murshid, Yoshihito Ishihara, Naoko Kajimura, Toshiaki Hasegawa, Ryoko Ando, Yasuyo Sugawara, Takashi Yamashiro, Akio Takaoka, Teruko Takano-Yamamoto

Research output: Contribution to journalArticle

41 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)377-383
Number of pages7
JournalMicroscopy and microanalysis : the official journal of Microscopy Society of America, Microbeam Analysis Society, Microscopical Society of Canada
Volume15
Issue number5
DOIs
Publication statusPublished - Oct 2009

ASJC Scopus subject areas

  • Instrumentation

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