Three-Dimensional Analysis of Melanosomes Isolated from B16 Melanoma Cells by Using Ultra High Voltage Electron Microscopy

• 作者: Akio Takaoka Hirotarou Mori Hitomi Aoki Huayua Chen Kenzo Koike Shuuichi Akazaki Takahiro Kunisada Tomoki Nishida Toshie Takahashi Yujiro Nakano
• 来源期刊: 显微镜研究(英文)
• 学科: 医学
• 关键词: B16 MELANOMA MELANOSOME SCANNING ELECTRON MICROSCOPY Transmission ELECTRON MICROSCOPY Ultra High Voltage ELECTRON MICROSCOPY Three-Dimensional Structure
• 年，卷（期）: 2014,（1）
• 页码范围: 1-8
• 页数: 8页
• 分类号: R73

摘要

Melanosomes, isolated by centrifugal separation from culture broth of B16 melanoma cells derived from mouse, were observed by scanning electron microscopy (SEM), and by transmission electron microscopy (TEM). Some interesting structural features were found inside and outside of the melanosomes. By SEM observation, the melanosomes were ellipsoid shape, their surface was not smooth and was covered with rough substructure, 10 to 20 nm particles. By TEM, uneven structure and micro particles were observed in the melanosomes. Furthermore, three-dimensional analysis was tried by using the ultra-high voltage electron microscopy(UHVEM). Micrographs of the melanosomes were taken at various tilted angles by UHVEM, after preparing 500 nm thickness specimens stained with lead citrate. From the micrographs collected, the three-dimensional structures were reconstructed by using i-mode software. Melanin stained by lead and non stained parts was clearly observed in the reconstructed structure. Non stained parts were round, regular size, and distributed widely in the melanosomes.