Tuning of the Zernike phase-plate for visualization of detailed ultrastructure in complex biological specimens

Fukuda Y, Fukazawa Y, Danev R, Shigemoto R, Nagayama K. 2009. Tuning of the Zernike phase-plate for visualization of detailed ultrastructure in complex biological specimens. Journal of Structural Biology. 168(3), 476–484.

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Journal Article | Published
Author
Fukuda, Yoshiyuki; Fukazawa, Yugo; Danev, Radostin S; Shigemoto, RyuichiIST Austria ; Nagayama, Kuniaki
Abstract
In order to acquire phase-contrast images with adequate contrast, conventional TEM requires large amount of defocus. Increasing the defocus improves the low-frequency components but attenuates the high-frequency ones. On the other hand, Zernike phase-contrast TEM (ZPC-TEM) can recover low-frequency components without losing the high-frequency ones under in-focus conditions. ZPC-TEM however, has another problem, especially in imaging of complex biological specimens such as cells and tissues; strong halos appear around specimen structures, and these halos hinder the interpretation of images. Due to this problem, the application of ZPC-TEM has been restricted to imaging of smaller particles. In order to improve the halo appearance, we fabricated a new quarter-wave thin film phase-plate with a smaller central hole and tested it on vitreous biological specimens. ZPC-TEM with the new plate could successfully visualize, in in-focus images, the intracellular fine features of cultured cells and brain tissues. This result indicates that reduction of the central hole diameter makes ZPC-TEM applicable on size scales ranging from protein particles to tissue sections. The application of ZPC-TEM to vitreous biological specimens will be a powerful method to advance the new field of imaging science for ultrastructures in close-to-physiological state.
Publishing Year
Date Published
2009-12-01
Journal Title
Journal of Structural Biology
Volume
168
Issue
3
Page
476 - 484
IST-REx-ID

Cite this

Fukuda Y, Fukazawa Y, Danev R, Shigemoto R, Nagayama K. Tuning of the Zernike phase-plate for visualization of detailed ultrastructure in complex biological specimens. Journal of Structural Biology. 2009;168(3):476-484. doi:10.1016/j.jsb.2009.08.011
Fukuda, Y., Fukazawa, Y., Danev, R., Shigemoto, R., & Nagayama, K. (2009). Tuning of the Zernike phase-plate for visualization of detailed ultrastructure in complex biological specimens. Journal of Structural Biology. Academic Press. https://doi.org/10.1016/j.jsb.2009.08.011
Fukuda, Yoshiyuki, Yugo Fukazawa, Radostin Danev, Ryuichi Shigemoto, and Kuniaki Nagayama. “Tuning of the Zernike Phase-Plate for Visualization of Detailed Ultrastructure in Complex Biological Specimens.” Journal of Structural Biology. Academic Press, 2009. https://doi.org/10.1016/j.jsb.2009.08.011.
Y. Fukuda, Y. Fukazawa, R. Danev, R. Shigemoto, and K. Nagayama, “Tuning of the Zernike phase-plate for visualization of detailed ultrastructure in complex biological specimens,” Journal of Structural Biology, vol. 168, no. 3. Academic Press, pp. 476–484, 2009.
Fukuda Y, Fukazawa Y, Danev R, Shigemoto R, Nagayama K. 2009. Tuning of the Zernike phase-plate for visualization of detailed ultrastructure in complex biological specimens. Journal of Structural Biology. 168(3), 476–484.
Fukuda, Yoshiyuki, et al. “Tuning of the Zernike Phase-Plate for Visualization of Detailed Ultrastructure in Complex Biological Specimens.” Journal of Structural Biology, vol. 168, no. 3, Academic Press, 2009, pp. 476–84, doi:10.1016/j.jsb.2009.08.011.

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