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

Hyun Wook Kim, Namkug Kim, Ki Woo Kim, Im Joo Rhyu

Research output: Contribution to journalArticle

4 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)172-177
Number of pages6
JournalMicroscopy and Microanalysis
Volume19
Issue numberSUPPL. 5
DOIs
Publication statusPublished - 2013 Aug 1

Fingerprint

abrasion
Abrasion
cerebellum
Imaging techniques
Scanning electron microscopy
scanning electron microscopy
fibers
Fibers
Focused ion beams
Ions
computer programs
mice
ions
ion beams
nervous system
synapses
Neurology
Field emission
field emission
phosphates

Keywords

  • cerebellum
  • ion-abrasion SEM
  • mossy fiber rosette
  • three dimension

ASJC Scopus subject areas

  • Instrumentation

Cite this

Three-dimensional imaging of cerebellar mossy fiber rosettes by ion-abrasion scanning electron microscopy. / Kim, Hyun Wook; Kim, Namkug; Kim, Ki Woo; Rhyu, Im Joo.

In: Microscopy and Microanalysis, Vol. 19, No. SUPPL. 5, 01.08.2013, p. 172-177.

Research output: Contribution to journalArticle

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