<Originals>Immuno-scanning electron microscopy of the macrophage cytoskeletons
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概要
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Mouse peritoneal macrophages (Mφ) were treated with dimethyl-3,3'-dithiobispropionimidate dihydrochloride, Triton X-100,and then KCl plus Triton X-100 (D-T-K method), and their cytoskeletons were observed by scanning electron microscopy (SEM). The secondary electron images (SEI) of the treated cells showed a radial extension of cytoskeletal filaments from the nucleus to the periphery of the cell, presenting a three-dimensional meshwork. Immunoenzymatic light microscopy showed that actin filament bundles, microtubules and vimentin filament bundles widely distributed, suggesting that the Mφ treated by the D-T-K method retained three kinds of cytoskeletal filaments with their antigenicity. The SEI of the cultured cells (P388D1,HeLa S-3 and L929) treated by the present method also revealed a clear topographical organization of cytoskeletons. To identify each filament by SEM, the author used antibodies to actin, tubulin or vimentin and then secondary antibodies labeled with colloidal gold particles. In the SEI, it was difficult to distinguish the gold markers from unknown cellular structures of marker size. However, backscattered electron images (BEI : composition images), especially their stereomicrographs clearly showed the gold particles attached to some of the cytoskeletal filaments. The author could identify three kinds of Mφ cytoskeletons, namely microfilaments, microtubules and vimentin filaments by immuno-SEM. The most prevalent component of the cytoplasmic cytoskeletons was microfilaments.
- 近畿大学の論文