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Journal of Electron Microscopy Advance Access published online on June 28, 2006
Journal of Electron Microscopy, doi:10.1093/jmicro/dfl016
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© The Author 2006. Published by Oxford University Press on behalf of Japanese Society of Microscopy. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org
Received January 5, 2006
Accepted May 15, 2006

Full-length paper: Biological

Observations of unstained biological specimens using a low-energy, high-resolution STEM

Akio Takaoka 1 * and Toshiaki Hasegawa 1

1 Research Center for Ultra-High Voltage Electron Microscopy, Osaka University, Mihoga-oka 7-1, Ibaraki, Osaka 567-0047, Japan

* To whom correspondence should be addressed.
Akio Takaoka, E-mail: takaoka{at}uhvem.osaka-u.ac.jp


  Abstract

Low-energy, high-resolution scanning transmission electron microscopy (STEM) is introduced as a convenient method for observing unstained biological specimens. By reducing the electron energy, the cross section for light elements becomes comparable to that of conventional electron microscopy observations. The STEM mode exhibited the advantage that the induced energy loss and charge build-up in the sample affected the image to a lesser extent than in the TEM or SEM mode. Furthermore, the efficiency of an STEM detector is high, and the total radiation damage can be reduced if thermal damage due to localized heating at a slow scan operation can be overcome. We applied this method for observations of biological samples that were in the form of thin slices, fine fibers and small particles. When the supporting film for samples is absent, the resolution and the contrast of STEM images can be maintained similar to SEM and TEM images, respectively.

Keywords: STEM; unstained specimen; low energy; cross section; chromatin fiber.
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