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Journal of Electron Microscopy Advance Access published online on October 21, 2008
Journal of Electron Microscopy, doi:10.1093/jmicro/dfn020
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© The Author 2008. Published by Oxford University Press on behalf of Japanese Society of Microscopy. All rights reserved. For permissions, please e-mail: journals.permissions@oxfordjournals.org

A new device for high-pressure freezing of cultured cell monolayer using 10-µm-thin stainless discs as both culture plate and specimen carrier

Akira Sawaguchi1,*, Fumiyo Aoyama1, Soyuki Ide1, Yoshiteru Goto2 and Tatsuo Suganuma1

1 Department of Anatomy, Ultrastructural Cell Biology, Faculty of Medicine and
2 Division of Electron Microscopy, Frontier Science Research Center, University of Miyazaki, Japan

* To whom correspondence should be addressed. E-mail: sawa{at}fc.miyazaki-u.ac.jp

High-pressure freezing (HPF) has been generally accepted as the most reliable method for cryofixation of biological samples, yielding a deep vitreous freezing. In recent cell biology, mammalian cultured cells are widely used, but HPF of cultured cell monolayer has not reached its full potential. In this study, we developed a new reliable device for HPF of cultured cell monolayer by using a 10-µm-thin stainless disc both as culture plate and specimen carrier. We describe the practical procedure, and demonstrate fine structures of HeLa cells cultured and cryofixed on the stainless discs as results.

Keywords     high-pressure freezing, cultured cell, monolayer, stainless disc

Received      8 July 2008, accepted 25 September 2008


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