@article{oai:ir.kagoshima-u.ac.jp:00011477,
 author = {Sonomura, Takahiro and 薗村, 貴弘 and Furuta, Takahiro and 古田, 貴寛 and Nakatani, Ikuko and 中谷, 郁子 and Yamamoto, Yo and 山本, 洋 and Unzai, Tomo and 雲財, 知 and Matsuda, Wakoto and 松田, 和郎 and Iwai, Haruki and 岩井, 治樹 and Yamanaka, Atsushi and 山中, 淳之 and Uemura, Masanori and 植村, 正憲 and Kaneko, Takeshi and 金子, 武嗣},
 issue = {26},
 journal = {Frontiers in Neural Circuits},
 month = {},
 note = {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.},
 pages = {1--7},
 title = {Correlative analysis of immunoreactivity in confocal laser-scanning microscopy and scanning electron microscopy with focused ion beam milling},
 volume = {7},
 year = {2013},
 yomi = {ソノムラ, タカヒロ and フルタ, タカヒロ and ナカタニ, イクコ and ヤマモト, ヨウ and ウンザイ, トモ and マツダ, ワコト and イワイ, ハルキ and ヤマナカ, アツシ and ウエムラ, マサノリ and カネコ, タケシ}
}