A modified 'NanoSuit((R))' preserves wet samples in high vacuum: direct observations on cells and tissues in field-emission scanning electron microscopy

作者：Takaku Yasuharu; Suzuki Hiroshi; Kawasaki Hideya; Ohta Isao; Ishii Daisuke; Hirakawa Satoshi; Tsutsui Takami; Matsumoto Haruko; Takehara Sayuri; Nakane Chinatsu; Sakaida Kana; Suzuki Chiaki; Muranaka Yoshinori; Kikuchi Hirotoshi; Konno Hiroyuki; Shimomura Masatsugu; Hariyama Takahiko^*

来源：Royal Society Open Science, 2017, 4(3): 160887.

DOI：10.1098/rsos.160887

• field-emission scanning electron microscope (FE-SEM)
• high vacuo
• living specimen
• surface shield effect
• NanoSuit((R))

Although field-emission scanning electron microscopy (FESEM) has proven very useful in biomedical research, the high vacuum required (10(-3) to 10(-7) Pa) 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 2017 The Authors. Published by the Royal Society under the terms of the Creative Commons Attribution License http://creativecommons. org/licenses/by/4.0/, which permits unrestricted use, provided the original author and source are credited. organisms in a thin, vacuum-proof suit, the 'NanoSuit((R))'. 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.

• 出版日期2017-3