title: Novel scanning electron microscopy methods for analyzing the 3D structure of the Golgi apparatus.
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.
authors: Department of Microscopic Anatomy and Cell Biology, Asahikawa Medical University, Midorigaoka Higashi 2-1-1-1, Asahikawa, 078-8510, Japan. daisukek@asahikawa-med.ac.jp.|Division of Microscopic Anatomy and Bio-imaging, Niigata University Graduate School of Medical and Dental Sciences, Niigata, Japan. daisukek@asahikawa-med.ac.jp.
authors: Daisuke
authors: D
authors: Koga
authors: Division of Microscopic Anatomy and Bio-imaging, Niigata University Graduate School of Medical and Dental Sciences, Niigata, Japan.
authors: Tatsuo
authors: T
authors: Ushiki
authors: Department of Microscopic Anatomy and Cell Biology, Asahikawa Medical University, Midorigaoka Higashi 2-1-1-1, Asahikawa, 078-8510, Japan.
authors: Tsuyoshi
authors: T
authors: Watanabe
mesh_terms: D000818:Animals; D018870:Endoplasmic Reticulum, Rough / Q000648:ultrastructure; D006056:Golgi Apparatus / Q000648:ultrastructure*; D052681:Gonadotrophs / Q000166:cytology / Q000648:ultrastructure; D021621:Imaging, Three-Dimensional / Q000379:methods*; D008855:Microscopy, Electron, Scanning / Q000379:methods*; D008928:Mitochondria / Q000648:ultrastructure; D009992:Osmium; D051381:Rats
chemical_list: D009992:Osmium
keywords: 3D; Array tomography using SEM; Golgi apparatus; Osmium maceration method; SEM