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Practical Experience with Hole-Free Phase Plates for Cryo Electron Microscopy

  • Michael Marko (a1) (a2), Chyongere Hsieh (a1), Eric Leith (a3), David Mastronarde (a4) and Sohei Motoki (a5)...

Abstract

Phase plate (PP) imaging has proven to be valuable in transmission cryo electron microscopy of unstained, native-state biological specimens. Many PP types have been described, however until the recent implementation of the “hole-free” phase plate (HFPP), imaging has been challenging. We found the HFPP to be simple to construct and to set up in the transmission electron microscopy, but care in implementing automated data collection is needed. Performance may be variable, both initially and over time, thus it is important to monitor and evaluate image quality by observing the power spectrum. We found that while some HFPPs gave transfer to high resolution without CTF oscillation, most reached high resolution when operated with modest defocus.

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Corresponding author

* Corresponding author. mike.marko.em@gmail.com

References

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Danev, R. & Baumeister, W. (2016). Cryo-EM single particle analysis with the Volta phase plate. Elife, 5. pii: e13046. doi: 10.7554/eLife.13046.
Danev, R., Buijsse, B., Khosouei, M., Plitzko, J.M. & Baumeister, W. (2014). Volta potential phase plate for in-focus phase contrast transmission electron microscopy. Proc Nat Acad Sci U S A 111, 1563515640.
Danev, R., Glaeser, R.M. & Nagayama, K. (2009). Practical factors affecting the performance of a thin-film phase plate for transmission electron microscopy. Ultramicroscopy 109, 312325.
Glaeser, R.M. (2013). Methods for imaging weak-phase objects in electron microscopy. Rev Sci Instr 85, 111101111117.
Hu, B., Margolin, W., Molineux, I.J. & Liu, J. (2015). Structural remodeling of bacteriophage T4 and host membranes during infection initiation. Proc Nat Acad Sci U S A 112, E4919E4928.
Mahamid, J., Pfeffer, S., Schaffer, M., Villa, E., Danev, R., Cuellar, L.K., Förster, F., Hyman, A.A., Plitzko, J.M. & Baumeister, W. (2016). Visualizing the molecular sociology at the HeLa cell nuclear periphery. Science 351, 969972.
Malac, M., Beleggia, M., Kawasaki, M., Li, P. & Egerton, R.F. (2012). Convenient contrast enhancement by a hole-free phase plate. Ultramicroscopy 118, 7789.
Marko, M., Leith, A., Hsieh, C. & Danev, R. (2011). Retrofit implementation of Zernike phase plate imaging for cryo-TEM. J Struct Biol 174, 400412.
Marko, M., Meng, X., Hsieh, C., Roussie, J. & Striemer, C. (2013). Methods for testing Zernike phase plates and a report on silicon-based phase plates with reduced charging and improved ageing characteristics. J Struct Biol 184, 237244.
Mastronarde, D.N. (1997). Dual-axis tomography: an approach with alignment methods that preserve resolution. J Struct Biol 120, 343352.
Mastronarde, D.N. (2005). Automated electron microscope tomography using robust prediction of specimen movements. J Struct Biol 152, 3651.

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Practical Experience with Hole-Free Phase Plates for Cryo Electron Microscopy

  • Michael Marko (a1) (a2), Chyongere Hsieh (a1), Eric Leith (a3), David Mastronarde (a4) and Sohei Motoki (a5)...

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