title: A modified 'NanoSuit®' preserves wet samples in high vacuum: direct observations on cells and tissues in field-emission scanning electron microscopy.
abstract: Although field-emission scanning electron microscopy (FE-SEM) has proven very useful in biomedical research, the high vacuum required (10-3to 10-7Pa) precludes direct observations of living cells and tissues at high resolution and often produces unwanted structural changes. We have previously described a method that allows the investigator to keep a variety of insect larvae alive in the high vacuum environment of the electron microscope by encasing the organisms in a thin, vacuum-proof suit, the 'NanoSuit®'. However, it was impossible to protect wet tissues freshly excised from intact organisms or cultured cells. Here we describe an improved 'NanoSuit' technique to overcome this limitation. We protected the specimens with a surface shield enhancer (SSE) solution that consists of glycerine and electrolytes and found that the fine structure of the SSE-treated specimens is superior to that of conventionally prepared specimens. The SSE-based NanoSuit affords a much stronger barrier to gas and/or liquid loss than the previous NanoSuit did and, since it allows more detailed images, it could significantly help to elucidate the 'real' organization of cells and their functions.
authors.affiliation: Department of Biology , Hamamatsu University School of Medicine , Higashi-ku, Hamamatsu 431-3192 , Japan.
authors.forename: Yasuharu
authors.initials: Y
authors.lastname: Takaku
authors.affiliation: Department of Chemistry , Hamamatsu University School of Medicine , Higashi-ku, Hamamatsu 431-3192 , Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Suzuki
authors.affiliation: Department of Regenerative and Infectious Pathology , Hamamatsu University School of Medicine , Higashi-ku, Hamamatsu 431-3192 , Japan.
authors.forename: Hideya
authors.initials: H
authors.lastname: Kawasaki
authors.affiliation: Laboratory for Ultrastructure Research and Research Equipment Center , Hamamatsu University School of Medicine , Higashi-ku, Hamamatsu 431-3192 , Japan.
authors.forename: Isao
authors.initials: I
authors.lastname: Ohta
authors.affiliation: Life Science and Applied Chemistry, Graduate School of Engineering , Nagoya Institute of Technology , Gokiso-cho, Showa-ku, Nagoya 466-8555 , Japan.
authors.forename: Daisuke
authors.initials: D
authors.lastname: Ishii
authors.affiliation: Department of Dermatology , Hamamatsu University School of Medicine , Higashi-ku, Hamamatsu 431-3192 , Japan.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Hirakawa
authors.affiliation: Department of Biology , Hamamatsu University School of Medicine , Higashi-ku, Hamamatsu 431-3192 , Japan.
authors.forename: Takami
authors.initials: T
authors.lastname: Tsutsui
authors.affiliation: Department of Biology , Hamamatsu University School of Medicine , Higashi-ku, Hamamatsu 431-3192 , Japan.
authors.forename: Haruko
authors.initials: H
authors.lastname: Matsumoto
authors.affiliation: Department of Biology , Hamamatsu University School of Medicine , Higashi-ku, Hamamatsu 431-3192 , Japan.
authors.forename: Sayuri
authors.initials: S
authors.lastname: Takehara
authors.affiliation: Department of Biology , Hamamatsu University School of Medicine , Higashi-ku, Hamamatsu 431-3192 , Japan.
authors.forename: Chinatsu
authors.initials: C
authors.lastname: Nakane
authors.affiliation: Department of Biology , Hamamatsu University School of Medicine , Higashi-ku, Hamamatsu 431-3192 , Japan.
authors.forename: Kana
authors.initials: K
authors.lastname: Sakaida
authors.affiliation: Department of Biology , Hamamatsu University School of Medicine , Higashi-ku, Hamamatsu 431-3192 , Japan.
authors.forename: Chiaki
authors.initials: C
authors.lastname: Suzuki
authors.affiliation: Department of Biology , Hamamatsu University School of Medicine , Higashi-ku, Hamamatsu 431-3192 , Japan.
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Muranaka
authors.affiliation: Second Department of Surgery , Hamamatsu University School of Medicine , Higashi-ku, Hamamatsu 431-3192 , Japan.
authors.forename: Hirotoshi
authors.initials: H
authors.lastname: Kikuchi
authors.affiliation: Second Department of Surgery , Hamamatsu University School of Medicine , Higashi-ku, Hamamatsu 431-3192 , Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Konno
authors.affiliation: Departments of Bio- and Material Photonics , Chitose Institute of Science and Technology , Hokkaido 066-8655 , Japan.
authors.forename: Masatsugu
authors.initials: M
authors.lastname: Shimomura
authors.affiliation: Department of Biology , Hamamatsu University School of Medicine , Higashi-ku, Hamamatsu 431-3192 , Japan.
authors.forename: Takahiko
authors.identifier: 0000-0001-9623-1011
authors.initials: T
authors.lastname: Hariyama
keywords: NanoSuit®; field-emission scanning electron microscope (FE-SEM); high vacuo; living specimen; surface shield effect