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Journal of Electron Microscopy Advance Access originally published online on February 12, 2009
Journal of Electron Microscopy 2009 58(2):35-45; doi:10.1093/jmicro/dfp004
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© The Author 2009. Published by Oxford University Press on behalf of Japanese Society of Microscopy. All rights reserved. For permissions, please e-mail: journals.permissions@oxfordjournals.org

High-contrast imaging of plastic-embedded tissues by phase contrast electron microscopy

Kimie Atsuzawa1, Nobuteru Usuda1, Ayami Nakazawa1, Motoaki Fukasawa1, Radostin Danev2, Shouzou Sugitani3 and Kuniaki Nagayama2,*

1 Department of Anatomy, School of Medicine, Fujita Health University, Toyoake, Aichi 470-1192
2 Okazaki Institute for Integrative Bioscience, National Institutes of Natural Sciences
3 Tera Base Co., Okazaki, Aichi 444-8787, Japan

* To whom correspondence should be addressed. E-mail: nagayama{at}nips.ac.jp

Phase contrast electron microscopy utilizing phase plates has been considered suitable for high-contrast observation of weak phase objects. This novel technique was newly applied to histochemically stained strong phase objects of osmificated biological specimens. Sections of various thicknesses, specifically stained for the Golgi apparatus by the ZIO technique using the heavy metals Zn and Os, were observed with a phase contrast electron microscope in Zernike and Hilbert imaging modes. Quantitative analysis of image contrast in real space and the power spectrum in Fourier space showed a high-contrast gain even for strong phase objects. This result clearly indicates that phase contrast electron microscopy can be effectively used not only for weak phase objects but also for strong phase objects in biology.

Keywords     phase contrast electron microscopy, phase plate, histochemistry, Golgi apparatus, power spectrum, strong phase object

Received      6 September 2008, accepted 15 January 2009


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