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Correlative analysis of immunoreactivity in confocal laser-scanning microscopy and scanning electron microscopy with focused ion beam milling
http://hdl.handle.net/10232/18518
http://hdl.handle.net/10232/185181ff2fa2e-87b8-4b16-be58-d06100e661dd
名前 / ファイル | ライセンス | アクション |
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SONOMURA_fncir_v7_26.pdf (1.3 MB)
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Item type | 学術雑誌論文 / Journal Article(1) | |||||
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公開日 | 2014-12-22 | |||||
タイトル | ||||||
タイトル | Correlative analysis of immunoreactivity in confocal laser-scanning microscopy and scanning electron microscopy with focused ion beam milling | |||||
言語 | en | |||||
著者 |
薗村, 貴弘
× 薗村, 貴弘× 古田, 貴寛× 中谷, 郁子× 山本, 洋× 雲財, 知× 松田, 和郎× 岩井, 治樹× 山中, 淳之× 植村, 正憲× 金子, 武嗣 |
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言語 | ||||||
言語 | eng | |||||
キーワード | ||||||
言語 | en | |||||
主題Scheme | Other | |||||
主題 | focused ion beam | |||||
キーワード | ||||||
言語 | en | |||||
主題Scheme | Other | |||||
主題 | scanning electron microscopy | |||||
キーワード | ||||||
言語 | en | |||||
主題Scheme | Other | |||||
主題 | immunocytochemistry | |||||
キーワード | ||||||
言語 | en | |||||
主題Scheme | Other | |||||
主題 | three-dimensional reconstruction | |||||
キーワード | ||||||
言語 | en | |||||
主題Scheme | Other | |||||
主題 | confocal laser-scanning microscope | |||||
キーワード | ||||||
言語 | en | |||||
主題Scheme | Other | |||||
主題 | neural circuit | |||||
キーワード | ||||||
言語 | en | |||||
主題Scheme | Other | |||||
主題 | synapse | |||||
キーワード | ||||||
言語 | en | |||||
主題Scheme | Other | |||||
主題 | neostriatum | |||||
資源タイプ | ||||||
資源タイプ識別子 | http://purl.org/coar/resource_type/c_6501 | |||||
資源タイプ | journal article | |||||
要約(Abstract) | ||||||
内容記述タイプ | Other | |||||
内容記述 | Recently, three-dimensional reconstruction of ultrastructure of the brain has been realized with minimal effort by using scanning electron microscopy (SEM) combined with focused ion beam (FIB) milling (FIB-SEM). Application of immunohistochemical staining in electron microscopy (EM) provides a great advantage in that molecules of interest are specifically localized in ultrastructures. Thus, we applied immunocytochemistry for FIB-SEM and correlated this immunoreactivity with that in confocal laser-scanning microcopy (CF-LSM). Dendrites of medium-sized spiny neurons in the rat neostriatum were visualized using a recombinant viral vector, which labeled the infected neurons with membrane-targeted GFP in a Golgi stain-like fashion. Moreover, the thalamostriatal afferent terminals were immunolabeled with Cy5 fluorescence for vesicular glutamate transporter 2 (VGluT2). After detection of the sites of terminals apposed to the dendrites by using CF-LSM, GFP and VGluT2 immunoreactivities were further developed for EM by using immunogold/silver enhancement and immunoperoxidase/diaminobenzidine (DAB) methods, respectively. In contrast-inverted FIB-SEM images, silver precipitations and DAB deposits were observed as fine dark grains and diffuse dense profiles, respectively, indicating that these immunoreactivities were as easily recognizable as those in the transmission electron microscopy (TEM) images. Furthermore, in the sites of interest, some appositions displayed synaptic specializations of an asymmetric type. Thus, the present method was useful in the three-dimensional analysis of immunocytochemically differentiated synaptic connections in the central neural circuit. | |||||
言語 | en | |||||
収録雑誌名 |
en : Frontiers in Neural Circuits 巻 7, 号 26, p. 1-7, 発行日 2013 |
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作成日 | ||||||
日付 | 2013-02-25 | |||||
日付タイプ | Issued | |||||
ISSN | ||||||
収録物識別子タイプ | EISSN | |||||
収録物識別子 | 16625110 | |||||
PubMed Id | ||||||
関連タイプ | isIdenticalTo | |||||
識別子タイプ | PMID | |||||
関連識別子 | 23443927 | |||||
DOI | ||||||
関連タイプ | isIdenticalTo | |||||
識別子タイプ | DOI | |||||
関連識別子 | https://doi.org/10.3389/fncir.2013.00026 | |||||
出版タイプ | ||||||
出版タイプ | VoR | |||||
出版タイプResource | http://purl.org/coar/version/c_970fb48d4fbd8a85 | |||||
NDC | ||||||
主題Scheme | NDC | |||||
主題 | 490 | |||||
公開者・出版者 | ||||||
出版者 | Frontiers Research Foundation | |||||
言語 | en |