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Anatomical Science International
Published in: Anatomical Science International 1/2017

01-01-2017 | Review Article

Novel scanning electron microscopy methods for analyzing the 3D structure of the Golgi apparatus

Authors: Daisuke Koga, Tatsuo Ushiki, Tsuyoshi Watanabe

Published in: Anatomical Science International | Issue 1/2017

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Abstract

The structure of the Golgi apparatus has been extensively examined by light and electron microscopy, but details of its three-dimensional (3D) structure have remained unclear because of the technical limitations of conventional microscopy techniques. To overcome this problem, we have developed several novel scanning electron microscopy (SEM) methods for observing the 3D structure of subcellular organelles including the Golgi apparatus: (1) an osmium maceration method that facilitates SEM observation of membranous organelles, including the Golgi apparatus, by selectively removing soluble cytoplasmic proteins, (2) an osmium impregnation/maceration method that combines an osmium impregnation method with the osmium maceration method to determine the polarity of the Golgi apparatus by SEM, (3) a correlative light and SEM method that combines a cryosectioning technique with the osmium maceration method to enable correlation of the immunocytochemical distribution of molecules with the 3D ultrastructure of the Golgi apparatus, and (4) array tomography based on the systematic collection and integration of SEM images of serial ultrathin sections on glass slides for revealing the 3D ultrastructure of the entire Golgi apparatus. Together, the novel SEM techniques listed above can reveal the complete 3D structure of the Golgi apparatus in different cell types.
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Metadata
Title
Novel scanning electron microscopy methods for analyzing the 3D structure of the Golgi apparatus
Authors
Daisuke Koga
Tatsuo Ushiki
Tsuyoshi Watanabe
Publication date
01-01-2017
Publisher
Springer Japan
Published in
Anatomical Science International / Issue 1/2017
Print ISSN: 1447-6959
Electronic ISSN: 1447-073X
DOI
https://doi.org/10.1007/s12565-016-0380-8

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