Dressing living organisms in a thin polymer membrane, the NanoSuit, for high-vacuum FE-SEM observation.
Scanning electron microscopy (SEM) has made remarkable progress and has become an essential tool for observing biological materials at microscopic level. However, various complex procedures have precluded observation of living organisms to date. Here, a new method is presented by which living organisms can be observed by field emission (FE)-SEM. Using this method, active movements of living animals were observed in vacuo (10(-5)-10(-7) Pa) by protecting them with a coating of thin polymer membrane, a NanoSuit, and it was found that the surface fine structure of living organisms is very different from that of traditionally fixed samples. After observation of mosquito larvae in the high vacuum of the FE-SEM, it was possible to rear them subsequently in normal culture conditions. This method will be useful for numerous applications, particularly for electron microscopic observations in the life sciences.
Research Equipment Center.
Isao
I
Ohta
Department of Biology CREST, Japan Science and Technology Agency, Hon-cho 4-1-8, Kawaguchi 332-0012, Japan.
Yasuharu
Y
Takaku
Department of Chemistry, Hamamatsu University School of Medicine, 1-20-1 Handayama, Higashi-ku, Hamamatsu 431-3192, Japan.
Hiroshi
H
Suzuki
Center for Fostering Young and Innovative Researchers, Nagoya Institute of Technology, Gokiso-cho, Showa-ku, Nagoya 466-8555, Japan CREST, Japan Science and Technology Agency, Hon-cho 4-1-8, Kawaguchi 332-0012, Japan.
Daisuke
D
Ishii
Department of Biology.
Yoshinori
Y
Muranaka
Institute of Multidisciplinary Research for Advanced Materials (IMRAM), Tohoku University, 2-1-1 Katahira, Aoba-ku, Sendai 980-8577, Japan CREST, Japan Science and Technology Agency, Hon-cho 4-1-8, Kawaguchi 332-0012, Japan.
Masatsugu
M
Shimomura
Department of Biology CREST, Japan Science and Technology Agency, Hon-cho 4-1-8, Kawaguchi 332-0012, Japan hariyama@hama-med.ac.jp.
Takahiko
T
Hariyama
D033304:Amphipoda / Q000166:cytology*; D000818:Animals; D002683:Chironomidae / Q000166:cytology*; D021621:Imaging, Three-Dimensional / Q000379:methods; D007814:Larva / Q000166:cytology*; D008567:Membranes, Artificial; D008855:Microscopy, Electron, Scanning / Q000379:methods*; D011108:Polymers; D013048:Specimen Handling / Q000379:methods
D008567:Membranes, Artificial; D011108:Polymers
NanoSuit; field emission scanning electron microscope; high vacuo; living organism; surface-shield-effect