STUDY OF WHOLE-MOUNT CYTOSKELETON OF HUMAN GLIOBLASTOMA BT_(325) CELLS BY CARBONPLATINUM COATING AND IMMUNOELECTRON MICROSCOPY
- VernacularTitle:用碳-铂复型与免疫电镜技术对人脑胶质瘤细胞系BT_(325)整装细胞骨架的研究
- Author:
Zhengmei LIU
;
Xibin LI
- Publication Type:Journal Article
- Keywords:
Cytoskeleton;
Human glioblastoma cell line BT_(325);
Carbonplatinum replica;
Immunoelectron microscopy
- From:
Acta Anatomica Sinica
1953;0(01):-
- CountryChina
- Language:Chinese
-
Abstract:
Cells of BT_(325), a human glioblastoma cell line, were treated with 0.5% Triton X-100 and immunostained with a monoclonal antibody to vimentin and antiserum to glial fibrillary acidic protein (GFAP), and followed by fluorescence labeled secondary antibody. After observation with fluorescence microscope the cells were dehydrated in increasing grades of ethanol and then critical point-dried. The carbon-platinum coating replicas of cytoskeleton were observed by transmission electron microscopy. The morphology of whole cytoskeleton as well as the arrangement of microtubules, microfilaments and intermediate filaments were clearly observed by this technique. It was observed that the diameter of intermediate filaments highly increased in the cells stained with the monoclonal antibody to vimentin. In the samples stained with anti-GFAP intermediate filaments were decorated in some cells but undecorated in other cells. The results indicates that all BT_(325) cells express vimentin, but only a fraction of the cells express GFAP. It was also observed that if the cells were fixed with formaldehyde before immunostaing with the monoclonal antibody to vimentin, the diameter of the intermediate filaments only slightly increased. However, the diameter of the intermediate filaments highly increaed in the cells stained firstly with the monoclonal antibody to vimentin and followed by formaldehyde fixation. This result is in accord with that of our previous immunofluorescence study. It indicates that formaldehyde fixation may "mask" certain epitopes on vimentin molecules.
