Phase Plate Electron Microscopy
An electron microscope enhanced with phase plates has practical advantages, particular for biological electron microscopy. It permits collecting high-contrast images of close-to-life biological structures with cryo-fixation and without harsh sample preparations including staining. Here we describe the state of the art of phase plate electron microscopy. Focuses are given to methodological aspects and biological applications with two dimensional and three-dimensional imaging.
KeywordsElectron microscopy Tomography Phase plate Phase contrast Thin-film Zernike phase contrast Hilbert differential contrast Cryo-fixation
We owe the development and biological applications of phase contrast TEM with phase plates to the collaborators as follows. Development: Radostin Danev, Shozo Sugitani, Hiroshi Okawara, Toshiyuki Itoh, Toshikazu Honda, Toshiaki Suzuki, Yoshihiro Arai, Fumio Hosokawa, Sohei Motoki, Rasmus Schroeder and Michael Marko. Applications: Yasuko Kaneko, Koji Nitta, Hitoshi Nakamoto, Nobutaru Usuda, Kimie Atsuzawa, Ayami Nakazawa, Kiyokazu Kametani, Masashi Yamaguchi and Mitsutoshi Setou. This work was supported in part by a Grant-in-aid for Creative Scientific Research from MEXT, Japan and by Core Research for Evolutional Science and Technology (CREST) from JST, Japan.
- Boersch, H. (1947) Über die Kontraste von Atomen in Electronenmikroskop. Z Naturforschg 2a: 615–633.Google Scholar
- Danev, R. and Nagayama, K. (2010) Phase plates for transmission electron microscopy. Meth. Enzymol. 481: 343–369.Google Scholar
- Danev, R., Kanamaru, S., Marko, M. and Nagayama, K. (2010) Zernike phase contrast cryo-electron tomography. J. Struct. Biol. 171(2): 174–181.Google Scholar
- Hosogi, N., Shigematsu, H., Terashima, H., Honma, M. and Nagayama, K. (2011) Zernike phase contrast cryo-electron tomography of sodium-driven flagellar hook-basal bodies from vibrio alginolytius. J. Struct. Biol. 173(1): 67–76.Google Scholar
- Kaneko, Y., Nitta, K. and Nagayama, K. (2007) Observation of in vivo DNA in ice embedded whole cyanobacterial cells by Hilbert differential contrast transmission electron microscopy (HDC-TEM). Plasma Fusion Res. 54: 79–85.Google Scholar
- Nitta, K., Nagayama, K., Danev, R. and Kaneko, Y. (2009) Visualization of BrdU-labelled DNA in cyanobacterial cells by Hilbert differential contrast transmission electron microscopy. 234: 118–123.Google Scholar
- Nomarski, G. (1952) Interferométre á polarization. French Patent 1: 059-123.Google Scholar
- Shiue, J., Chang, C.S., Huang, S.H., Hsu, C.H., Tsai, J.S., Chang, W.H., Wu, Y.M., Lin, Y.C., Kuo, P.C., Huang, Y.S., Hwu, Y., Kai, J.J., T. F.G. and Chen, F.R. (2009) Phase TEM for biological imaging utilizing a Boersh electrostatic phase plate: theory and practice. J. Electron Microsc. 58: 137–145.Google Scholar
- Smith, F.H. (1947) Microscopes. British Patent 639 014, Class 97(i) CroupXX.Google Scholar