TITLE:
Three-Dimensional Analysis of Melanosomes Isolated from B16 Melanoma Cells by Using Ultra High Voltage Electron Microscopy
AUTHORS:
Shuuichi Akazaki, Toshie Takahashi, Yujiro Nakano, Tomoki Nishida, Hirotarou Mori, Akio Takaoka, Hitomi Aoki, Huayua Chen, Takahiro Kunisada, Kenzo Koike
KEYWORDS:
B16 Melanoma; Melanosome; Scanning Electron Microscopy; Transmission Electron Microscopy; Ultra High Voltage Electron Microscopy; Three-Dimensional Structure
JOURNAL NAME:
Microscopy Research,
Vol.2 No.1,
January
22,
2014
ABSTRACT: 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.