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Immuno-electron microscopy of primary cell cultures from genetically modified animals in liquid by atmospheric scanning electron microscopy.
Kinoshita, Takaaki; Mori, Yosio; Hirano, Kazumi; Sugimoto, Shinya; Okuda, Ken-ichi; Matsumoto, Shunsuke; Namiki, Takeshi; Ebihara, Tatsuhiko; Kawata, Masaaki; Nishiyama, Hidetoshi; Sato, Mari; Suga, Mitsuo; Higashiyama, Kenichi; Sonomoto, Kenji; Mizunoe, Yoshimitsu; Nishihara, Shoko; Sato, Chikara.
Afiliación
  • Kinoshita T; 1 Laboratory of Cell Biology, Department of Bioinformatics, Faculty of Engineering, Soka University, 1-236 Tangi-machi, Hachioji, Tokyo 192-8577, Japan.
  • Mori Y; 2 Department of Virology III, National Institute of Infectious Diseases, 4-7-1 Gakuen, Musashimurayama-shi, Tokyo 208-0011, Japan.
  • Hirano K; 1 Laboratory of Cell Biology, Department of Bioinformatics, Faculty of Engineering, Soka University, 1-236 Tangi-machi, Hachioji, Tokyo 192-8577, Japan.
  • Sugimoto S; 3 Department of Bacteriology, The Jikei University School of Medicine, Tokyo 105-8461, Japan.
  • Okuda K; 3 Department of Bacteriology, The Jikei University School of Medicine, Tokyo 105-8461, Japan.
  • Matsumoto S; 4 Division of Structural Biology, Medical Institute of Bioregulation, Kyushu University, Fukuoka 812-8581, Japan.
  • Namiki T; 5 Suntory Global Innovation Center, Research Institute, 5-2-5 Yamazaki, Shimamoto-cho, Mishima-gun, Osaka 618-0001, Japan.
  • Ebihara T; 6 Biomedical Research Institute and Information Technology Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 1-1-1 Higashi, Tsukuba, Ibaraki 305-8566, Japan.
  • Kawata M; 6 Biomedical Research Institute and Information Technology Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 1-1-1 Higashi, Tsukuba, Ibaraki 305-8566, Japan.
  • Nishiyama H; 7 JEOL Ltd., 1-2 Musashino 3-chome, Akishima, Tokyo 196-8558, Japan.
  • Sato M; 6 Biomedical Research Institute and Information Technology Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 1-1-1 Higashi, Tsukuba, Ibaraki 305-8566, Japan.
  • Suga M; 7 JEOL Ltd., 1-2 Musashino 3-chome, Akishima, Tokyo 196-8558, Japan.
  • Higashiyama K; 5 Suntory Global Innovation Center, Research Institute, 5-2-5 Yamazaki, Shimamoto-cho, Mishima-gun, Osaka 618-0001, Japan.
  • Sonomoto K; 8 Laboratory of Microbial Technology, Department of Bioscience and Biotechnology, Division of Applied Molecular Microbiology and Biomass Chemistry, Faculty of Agriculture, Kyushu University, Fukuoka 812-8581, Japan.
  • Mizunoe Y; 3 Department of Bacteriology, The Jikei University School of Medicine, Tokyo 105-8461, Japan.
  • Nishihara S; 1 Laboratory of Cell Biology, Department of Bioinformatics, Faculty of Engineering, Soka University, 1-236 Tangi-machi, Hachioji, Tokyo 192-8577, Japan.
  • Sato C; 6 Biomedical Research Institute and Information Technology Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 1-1-1 Higashi, Tsukuba, Ibaraki 305-8566, Japan.
Microsc Microanal ; 20(2): 469-83, 2014 Apr.
Article en En | MEDLINE | ID: mdl-24564988
High-throughput immuno-electron microscopy is required to capture the protein-protein interactions realizing physiological functions. Atmospheric scanning electron microscopy (ASEM) allows in situ correlative light and electron microscopy of samples in liquid in an open atmospheric environment. Cells are cultured in a few milliliters of medium directly in the ASEM dish, which can be coated and transferred to an incubator as required. Here, cells were imaged by optical or fluorescence microscopy, and at high resolution by gold-labeled immuno-ASEM, sometimes with additional metal staining. Axonal partitioning of neurons was correlated with specific cytoskeletal structures, including microtubules, using primary-culture neurons from wild type Drosophila, and the involvement of ankyrin in the formation of the intra-axonal segmentation boundary was studied using neurons from an ankyrin-deficient mutant. Rubella virus replication producing anti-double-stranded RNA was captured at the host cell's plasma membrane. Fas receptosome formation was associated with clathrin internalization near the surface of primitive endoderm cells. Positively charged Nanogold clearly revealed the cell outlines of primitive endoderm cells, and the cell division of lactic acid bacteria. Based on these experiments, ASEM promises to allow the study of protein interactions in various complexes in a natural environment of aqueous liquid in the near future.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Animales Modificados Genéticamente / Microscopía Electrónica de Rastreo / Microscopía Inmunoelectrónica / Técnicas Citológicas / Drosophila Límite: Animals Idioma: En Revista: Microsc Microanal Año: 2014 Tipo del documento: Article País de afiliación: Japón Pais de publicación: Reino Unido
Texto completo
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PubMed Links
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Animales Modificados Genéticamente / Microscopía Electrónica de Rastreo / Microscopía Inmunoelectrónica / Técnicas Citológicas / Drosophila Límite: Animals Idioma: En Revista: Microsc Microanal Año: 2014 Tipo del documento: Article País de afiliación: Japón Pais de publicación: Reino Unido