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Biological application of focus ion beam-scanning electron microscopy (FIB-SEM) to the imaging of cartilaginous fibrils and osteoblastic cytoplasmic processes

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Title: Biological application of focus ion beam-scanning electron microscopy (FIB-SEM) to the imaging of cartilaginous fibrils and osteoblastic cytoplasmic processes
Authors: Hasegawa, Tomoka Browse this author
Endo, Takashi Browse this author
Tsuchiya, Erika Browse this author
Kudo, Ai Browse this author
Zhao, Shen Browse this author
Moritani, Yasuhito Browse this author
Abe, Miki Browse this author
Yamamoto, Tomomaya Browse this author
Hongo, Hiromi Browse this author
Tsuboi, Kanako Browse this author
Yoshida, Taiji Browse this author
Nagai, Tomoya Browse this author
Khadiza, Naznin Browse this author
Yokoyama, Ayako Browse this author
Luiz de Freitas, Paulo Henrique Browse this author
Li, Minqi Browse this author
Amizuka, Norio Browse this author →KAKEN DB
Keywords: FIB-SEM
Cartilage matrix
Issue Date: Feb-2017
Publisher: Elsevier
Journal Title: Journal of Oral Biosciences
Volume: 59
Issue: 1
Start Page: 55
End Page: 62
Publisher DOI: 10.1016/j.job.2016.11.004
Abstract: Objectives: The aim of this study is a biological application of focused ion beam-scanning electron microscopy (FIB-SEM) to demonstrate serial sectional images of skeletal tissues, here presenting the ultrastructure of 1) cartilaginous extracellular fibrils and 2) osteoblastic cytoplasmic processes. Methods: Seven weeks-old female wild-type mice were fixed with half-Karnovsky solution and subsequent OsO4, and the tibiae were extracted for block staining prior to observation under transmission electron microscope (TEM) and FIB-SEM. Results: TEM showed the fine fibrillar, but somewhat amorphous ultrastructure of the intercolumnar septa in the growth plate cartilage. Alternatively, FIB-SEM revealed bundles of stout fibrils at regular intervals paralleling the septa’s longitudinal axis, as well as vesicular structures embedded in the cartilaginous matrix of the proliferative zone. In the primary trabeculae, both TEM and FIB-SEM showed several osteoblastic cytoplasmic processes on the osteoid, with numbers higher than those seen in the bone matrix. FIB-SEM revealed the agglomeration of cytoplasmic processes beneath the osteoblasts, which formed a tubular continuum extending from those cells. Based on these findings, we postulated that osteoblasts not only extend their cytoplasmic processes through to the bone matrix, but also stack these cell processes on the osteoid of the primary trabeculae. Conclusion: Taken together, it is likely that FIB-SEM imaging strategy on serial sections may successfully deliver new insights on the ultrastructure of cartilage and bone tissues.
Rights: © 2017. This manuscript version is made available under the CC-BY-NC-ND 4.0 license
Type: article (author version)
Appears in Collections:歯学院・歯学研究院 (Graduate School of Dental Medicine / Faculty of Dental Medicine) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 長谷川 智香

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