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Three-Dimensional Imaging of Cerebellar Mossy Fiber Rosettes by Ion-Abrasion Scanning Electron Microscopy

Published online by Cambridge University Press:  06 August 2013

Hyun-Wook Kim ,
Namkug Kim ,
Ki Woo Kim  and
Im Joo Rhyu
Hyun-Wook Kim
Affiliation:
Department of Anatomy, College of Medicine, Korea University, 126-1 Anam dong 5 ka SungBuk Ku, Seoul 136-705, Korea
Namkug Kim
Affiliation:
Department of Radiology, Research Institute of Radiology, Asan Medical Center, University of Ulsan College of Medicine, 388-1 Pungnap-dong, Songpa-gu, Seoul 138-736, Korea
Ki Woo Kim*
Affiliation:
School of Ecology and Environmental System, Kyungpook National University, Sangju 742-711, Korea
Im Joo Rhyu*
Affiliation:
Department of Anatomy, College of Medicine, Korea University, 126-1 Anam dong 5 ka SungBuk Ku, Seoul 136-705, Korea
*
†Corresponding author. E-mail: kiwoo@knu.ac.kr
‡Corresponding author. E-mail: irhyu@korea.ac.kr
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Abstract

The detailed knowledge of the three-dimensional (3D) organization of the nervous tissue provides essential information on its functional elucidation. We used serial block-face scanning electron microscopy with focused ion beam (FIB) milling to reveal 3D morphologies of the mossy fiber rosettes in the mice cerebellum. Three-week-old C57 black mice were perfused with a fixative of 1% paraformaldehyde/1% glutaraldehyde in phosphate buffer; the cerebellum was osmicated and embedded in the Araldite. The block containing granule cell layer was sliced with FIB and observed by field-emission scanning electron microscopy. The contrast of backscattered electron image of the block-face was similar to that of transmission electron microscopy and processed using 3D visualization software for further analysis. The mossy fiber rosettes on each image were segmented and rendered to visualize the 3D model. The complete 3D characters of the mossy fiber rosette could be browsed on the A-Works, in-house software, and some preliminary quantitative data on synapse of the rosette could be extracted from these models. Thanks to the development of two-beam imaging and optimized software, we could get 3D information on cerebellar mossy fiber rosettes with ease and speedily, which would be an additive choice to explore 3D structures of the nervous systems and their networks.

Keywords

cerebellummossy fiber rosettethree dimensionion-abrasion SEM
Type
Research Article
Information
Microscopy and Microanalysis , Volume 19 , Supplement S5: IUMAS , August 2013 , pp. 172 - 177
Copyright
Copyright © Microscopy Society of America 2013 

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