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Journal of Electron Microscopy Advance Access originally published online on March 16, 2009
Journal of Electron Microscopy 2009 58(3):137-145; doi:10.1093/jmicro/dfp006
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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

This article appears in the following Journal of Electron Microscopy issue: Special number: Advanced electron microscopy in materials physics [View the issue table of contents]

Phase TEM for biological imaging utilizing a Boersch electrostatic phase plate: theory and practice

Jessie Shiue1, Chia-Seng Chang1, Sen-Hui Huang4, Chih-Hao Hsu4, Jin-Sheng Tsai3, Wei-Hau Chang2, Yi-Min Wu2, Yen-Chen Lin2, Pai-Chia Kuo1, Yang-Shan Huang1, Yeukuang Hwu1,3, Ji-Jung Kai4, Fan-Gang Tseng4 and Fu-Rong Chen1,3,4*

1 Institute of Physics
2 Institute of Chemistry, Academia Sinica, Nankang 11529, Taipei
3 National Synchrotron Radiation Research Center, HsinChu 30076
4 Department of Engineering and System Science, National TsingHua University, HsinChu 30043, Taiwan

* To whom correspondence should be addressed. E-mail: frchen{at}ess.nthu.edu.tw

A Boersch electrostatic phase plate (BEPP) used in a transmission electron microscope (TEM) system can provide tuneable phase shifts and overcome the low contrast problem for biological imaging. Theoretically, a pure phase image with a high phase contrast can be obtained using a BEPP. However, a currently available TEM system utilizing a BEPP cannot achieve sufficiently high phase efficiency for biological imaging, owing to the practical conditions. The low phase efficiency is a result of the blocking of partial unscattered electrons by BEPP, and the contribution of absorption contrast. The fraction of blocked unscattered beam is related to BEPP dimensions and to divergence of the illumination system of the TEM. These practical issues are discussed in this paper. Phase images of biological samples (negatively stained ferritin) obtained by utilizing a BEPP are reported, and the phase contrast was found to be enhanced by a factor of ~1.5, based on the calculation using the Rose contrast criterion. The low gain in phase contrast is consistent with the expectation from the current TEM/BEPP system. A new generation of phase TEM utilizing BEPP and designed for biological imaging with a high phase efficiency is proposed.

Keywords     phase TEM, Beorsch phase plate, Zernike phase plate, electrostatic phase plate, biological imaging, phase contrast

Received      2 December 2008, accepted 20 January 2009


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