title: Conductive resins improve charging and resolution of acquired images in electron microscopic volume imaging.
abstract: Recent advances in serial block-face imaging using scanning electron microscopy (SEM) have enabled the rapid and efficient acquisition of 3-dimensional (3D) ultrastructural information from a large volume of biological specimens including brain tissues. However, volume imaging under SEM is often hampered by sample charging, and typically requires specific sample preparation to reduce charging and increase image contrast. In the present study, we introduced carbon-based conductive resins for 3D analyses of subcellular ultrastructures, using serial block-face SEM (SBF-SEM) to image samples. Conductive resins were produced by adding the carbon black filler, Ketjen black, to resins commonly used for electron microscopic observations of biological specimens. Carbon black mostly localized around tissues and did not penetrate cells, whereas the conductive resins significantly reduced the charging of samples during SBF-SEM imaging. When serial images were acquired, embedding into the conductive resins improved the resolution of images by facilitating the successful cutting of samples in SBF-SEM. These results suggest that improving the conductivities of resins with a carbon black filler is a simple and useful option for reducing charging and enhancing the resolution of images obtained for volume imaging with SEM.
authors.affiliation: Department of Anatomy and Molecular Histology, Interdisciplinary Graduate School of Medicine and Engineering, University of Yamanashi, Chuo-city, Yamanashi 409-3898, Japan.
authors.forename: Huy Bang
authors.initials: HB
authors.lastname: Nguyen
authors.affiliation: Department of Anatomy and Molecular Histology, Interdisciplinary Graduate School of Medicine and Engineering, University of Yamanashi, Chuo-city, Yamanashi 409-3898, Japan.
authors.forename: Truc Quynh
authors.initials: TQ
authors.lastname: Thai
authors.affiliation: Department of Anatomy and Molecular Histology, Interdisciplinary Graduate School of Medicine and Engineering, University of Yamanashi, Chuo-city, Yamanashi 409-3898, Japan.
authors.forename: Sei
authors.initials: S
authors.lastname: Saitoh
authors.affiliation: Department of Anatomy and Molecular Histology, Interdisciplinary Graduate School of Medicine and Engineering, University of Yamanashi, Chuo-city, Yamanashi 409-3898, Japan.
authors.forename: Bao
authors.initials: B
authors.lastname: Wu
authors.affiliation: Department of Anatomy and Molecular Histology, Interdisciplinary Graduate School of Medicine and Engineering, University of Yamanashi, Chuo-city, Yamanashi 409-3898, Japan.
authors.forename: Yurika
authors.initials: Y
authors.lastname: Saitoh
authors.affiliation: Department of Occupational Therapy, Health Science University, Fujikawaguchiko, Yamanashi 401-0380, Japan.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Shimo
authors.affiliation: Gatan Japan, Koto-ku, Tokyo 135-0033, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Fujitani
authors.affiliation: Asahi Kasei Chemicals Corporation, Kawasaki-city, Kanagawa 210-0863 Japan.
authors.forename: Hirohide
authors.initials: H
authors.lastname: Otobe
authors.affiliation: Department of Anatomy and Molecular Histology, Interdisciplinary Graduate School of Medicine and Engineering, University of Yamanashi, Chuo-city, Yamanashi 409-3898, Japan.|Center for Multidisciplinary Brain Research, National Institute for Physiological Sciences, Okazaki, Aichi 444-8787, Japan.
authors.forename: Nobuhiko
authors.initials: N
authors.lastname: Ohno
mesh_terms: D000818:Animals; D001921:Brain / Q000648:ultrastructure; D004553:Electric Conductivity; D004853:Epoxy Resins / Q000737:chemistry*; D021621:Imaging, Three-Dimensional / Q000379:methods*; D007668:Kidney / Q000648:ultrastructure; D008810:Mice, Inbred C57BL; D008855:Microscopy, Electron, Scanning / Q000379:methods*; D015203:Reproducibility of Results; D053260:Soot / Q000737:chemistry*; D013048:Specimen Handling / Q000379:methods
chemical_list: D004853:Epoxy Resins; D053260:Soot