Three-dimensional imaging of cerebellar mossy fiber rosettes by ion-abrasion scanning electron microscopy

Kim, HW; Kim, N; Kim, KW; Rhyu, IJ

HERO ID

1997845

Reference Type

Journal Article

Year

2013

Language

English

PMID

23920200

HERO ID 1997845
In Press No
Year 2013
Title Three-dimensional imaging of cerebellar mossy fiber rosettes by ion-abrasion scanning electron microscopy
Authors Kim, HW; Kim, N; Kim, KW; Rhyu, IJ
Journal Microscopy and Microanalysis
Volume 19 Suppl 5
Issue S5
Page Numbers 172-177
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.
Doi 10.1017/S1431927613012609
Pmid 23920200
Wosid WOS:000332111100039
Url /www.cambridge.org
Is Certified Translation No
Dupe Override No
Comments Journal: Microscopy and microanalysis : the official journal of Microscopy Society of America, Microbeam Analysis Society, Microscopical Society of Canada ISSN: 1435-8115
Is Public Yes
Language Text English
Keyword Science & Technology - Other Topics; Chemistry; Instruments &; Instrumentation; Spectroscopy; cerebellum, mossy fiber rosette, three dimension, ion-abrasion SEM; caenorhabditis-elegans, synapses, beam, spines, rat, ltp,; reconstruction, dendrite