Practical aspects of Boersch phase contrast electron microscopy of biological specimens

Walter A, Muzik H, Vieker H, Turchanin A, Beyer A, Gölzhäuser A, Lacher M, Steltenkamp S, Schmitz S, Holik P, Kuehlbrandt W, et al. (2012)
Ultramicroscopy 116: 62-72.

Zeitschriftenaufsatz | Veröffentlicht | Englisch
Es wurden keine Dateien hochgeladen. Nur Publikationsnachweis!
Walter, Andreas; Muzik, HeikoUniBi; Vieker, HenningUniBi; Turchanin, AndreyUniBi; Beyer, AndréUniBi ; Gölzhäuser, ArminUniBi ; Lacher, Manfred; Steltenkamp, Siegfried; Schmitz, Sam; Holik, Peter; Kuehlbrandt, Werner; Rhinow, Daniel
Abstract / Bemerkung
Implementation of physical phase plates into transmission electron microscopes to achieve in-focus contrast for ice-embedded biological specimens poses several technological challenges. During the last decade several phase plates designs have been introduced and tested for electron ciyo-microscopy (cryoEM), including thin film (Zernike) phase plates and electrostatic devices. Boersch phase plates (BPPs) are electrostatic einzel lenses shifting the phase of the unscattered beam by an arbitrary angle. Adjusting the phase shift to 90 achieves the maximum contrast transfer for phase objects such as biomolecules. Recently, we reported the implementation of a BPP into a dedicated phase contrast aberration-corrected electron microscope (PACEM) and demonstrated its use to generate in-focus contrast of frozen-hydrated specimens. However, a number of obstacles need to be overcome before BPPs can be used routinely, mostly related to the phase plate devices themselves. CryoEM with a physical phase plate is affected by electrostatic charging, obliteration of low spatial frequencies, and mechanical drift. Furthermore, BPPs introduce single sideband contrast (SSB), due to the obstruction of Friedel mates in the diffraction pattern. In this study we address the technical obstacles in detail and show how they may be overcome. We use X-ray photoelectron spectroscopy (XPS) and Auger electron spectroscopy (AES) to identify contaminants responsible for electrostatic charging, which occurs with most phase plates. We demonstrate that obstruction of low-resolution features is significantly reduced by lowering the acceleration voltage of the microscope. Finally, we present computational approaches to correct BPP images for SSB contrast and to compensate for mechanical drift of the BPP. (C) 2012 Elsevier B.V. All rights reserved.
ion microscopy; Transmission electron microscopy; Helium; Phase plate; Aberration correction; Auger electron spectroscopy; X-ray photoelectron spectroscopy
Page URI


Walter A, Muzik H, Vieker H, et al. Practical aspects of Boersch phase contrast electron microscopy of biological specimens. Ultramicroscopy. 2012;116:62-72.
Walter, A., Muzik, H., Vieker, H., Turchanin, A., Beyer, A., Gölzhäuser, A., Lacher, M., et al. (2012). Practical aspects of Boersch phase contrast electron microscopy of biological specimens. Ultramicroscopy, 116, 62-72. doi:10.1016/j.ultramic.2012.03.009
Walter, A., Muzik, H., Vieker, H., Turchanin, A., Beyer, A., Gölzhäuser, A., Lacher, M., Steltenkamp, S., Schmitz, S., Holik, P., et al. (2012). Practical aspects of Boersch phase contrast electron microscopy of biological specimens. Ultramicroscopy 116, 62-72.
Walter, A., et al., 2012. Practical aspects of Boersch phase contrast electron microscopy of biological specimens. Ultramicroscopy, 116, p 62-72.
A. Walter, et al., “Practical aspects of Boersch phase contrast electron microscopy of biological specimens”, Ultramicroscopy, vol. 116, 2012, pp. 62-72.
Walter, A., Muzik, H., Vieker, H., Turchanin, A., Beyer, A., Gölzhäuser, A., Lacher, M., Steltenkamp, S., Schmitz, S., Holik, P., Kuehlbrandt, W., Rhinow, D.: Practical aspects of Boersch phase contrast electron microscopy of biological specimens. Ultramicroscopy. 116, 62-72 (2012).
Walter, Andreas, Muzik, Heiko, Vieker, Henning, Turchanin, Andrey, Beyer, André, Gölzhäuser, Armin, Lacher, Manfred, Steltenkamp, Siegfried, Schmitz, Sam, Holik, Peter, Kuehlbrandt, Werner, and Rhinow, Daniel. “Practical aspects of Boersch phase contrast electron microscopy of biological specimens”. Ultramicroscopy 116 (2012): 62-72.

7 Zitationen in Europe PMC

Daten bereitgestellt von Europe PubMed Central.

Biological Applications at the Cutting Edge of Cryo-Electron Microscopy.
Dillard RS, Hampton CM, Strauss JD, Ke Z, Altomara D, Guerrero-Ferreira RC, Kiss G, Wright ER., Microsc Microanal 24(4), 2018
PMID: 30175702
Towards an optimum design for thin film phase plates.
Rhinow D., Ultramicroscopy 160(), 2016
PMID: 26397752
Volta phase plate cryo-EM of the small protein complex Prx3.
Khoshouei M, Radjainia M, Phillips AJ, Gerrard JA, Mitra AK, Plitzko JM, Baumeister W, Danev R., Nat Commun 7(), 2016
PMID: 26817416
Towards an optimum design for electrostatic phase plates.
Walter A, Steltenkamp S, Schmitz S, Holik P, Pakanavicius E, Sachser R, Huth M, Rhinow D, Kühlbrandt W., Ultramicroscopy 153(), 2015
PMID: 25697462
Visualization of ATP synthase dimers in mitochondria by electron cryo-tomography.
Davies KM, Daum B, Gold VA, Mühleip AW, Brandt T, Blum TB, Mills DJ, Kühlbrandt W., J Vis Exp (91), 2014
PMID: 25285856
Volta potential phase plate for in-focus phase contrast transmission electron microscopy.
Danev R, Buijsse B, Khoshouei M, Plitzko JM, Baumeister W., Proc Natl Acad Sci U S A 111(44), 2014
PMID: 25331897

32 References

Daten bereitgestellt von Europe PubMed Central.

Optimizing phase contrast in transmission electron microscopy with an electrostatic (Boersch) phase plate.
Majorovits E, Barton B, Schultheiss K, Perez-Willard F, Gerthsen D, Schroder RR., Ultramicroscopy 107(2-3), 2006
PMID: 16949755
Design of a microfabricated, two-electrode phase-contrast element suitable for electron microscopy.
Cambie R, Downing KH, Typke D, Glaeser RM, Jin J., Ultramicroscopy 107(4-5), 2006
PMID: 17079082
Design of an electron microscope phase plate using a focused continuous-wave laser
Müller, New Journal of Physics 12(), 2010
Imaging of soft and hard materials using a Boersch phase plate in a transmission electron microscope
Alloyeau, Ultramicroscopy 110(), 2010
New electrostatic phase plate for phase-contrast transmission electron microscopy and its application for wave-function reconstruction.
Schultheiss K, Zach J, Gamm B, Dries M, Frindt N, Schroder RR, Gerthsen D., Microsc. Microanal. 16(6), 2010
PMID: 20946700
In-focus electron microscopy of frozen-hydrated biological samples with a Boersch phase plate.
Barton B, Rhinow D, Walter A, Schroder R, Benner G, Majorovits E, Matijevic M, Niebel H, Muller H, Haider M, Lacher M, Schmitz S, Holik P, Kuhlbrandt W., Ultramicroscopy 111(12), 2011
PMID: 22088444
Über die Kontraste von Atomen im Elektronenmikroskop
Boersch, Zeitschrift fuer Naturforschung A 2(), 1947
Transmission electron microscopy with Zernike phase plate
Danev, Ultramicroscopy 81(), 2001
Zernike phase contrast electron microscopy of ice-embedded influenza A virus.
Yamaguchi M, Danev R, Nishiyama K, Sugawara K, Nagayama K., J. Struct. Biol. 162(2), 2008
PMID: 18313941
Zernike phase contrast cryo-electron microscopy and tomography for structure determination at nanometer and subnanometer resolutions.
Murata K, Liu X, Danev R, Jakana J, Schmid MF, King J, Nagayama K, Chiu W., Structure 18(8), 2010
PMID: 20696391
Zernike phase contrast cryo-electron tomography of sodium-driven flagellar hook-basal bodies from Vibrio alginolyticus.
Hosogi N, Shigematsu H, Terashima H, Homma M, Nagayama K., J. Struct. Biol. 173(1), 2010
PMID: 20705140
Seeing the portal in herpes simplex virus type 1 B capsids.
Rochat RH, Liu X, Murata K, Nagayama K, Rixon FJ, Chiu W., J. Virol. 85(4), 2010
PMID: 21106752
The fabrication and application of Zernike electrostatic phase plate
Huang, Journal of Electron Microscopy 55(), 2007
Phase TEM for biological imaging utilizing a Boersch electrostatic phase plate: theory and practice.
Shiue J, Chang CS, Huang SH, Hsu CH, Tsai JS, Chang WH, Wu YM, Lin YC, Kuo PC, Huang YS, Hwu Y, Kai JJ, Tseng FG, Chen FR., J Electron Microsc (Tokyo) 58(3), 2009
PMID: 19289850
Object wave reconstruction by phase-plate transmission electron microscopy.
Gamm B, Dries M, Schultheiss K, Blank H, Rosenauer A, Schroder RR, Gerthsen D., Ultramicroscopy 110(7), 2010
PMID: 20189718
Zernike phase plate cryoelectron microscopy facilitates single particle analysis of unstained asymmetric protein complexes.
Chang WH, Chiu MT, Chen CY, Yen CF, Lin YC, Weng YP, Chang JC, Wu YM, Cheng H, Fu J, Tu IP., Structure 18(1), 2010
PMID: 20152149
Retrofit implementation of Zernike phase plate imaging for cryo-TEM.
Marko M, Leith A, Hsieh C, Danev R., J. Struct. Biol. 174(2), 2011
PMID: 21272647
Removal of self-assembled monolayers of alkanethiolates on gold by plasma cleaning
Raiber, Surface Science 595(), 2005

Hayashi, Journal of Catalysis 178(), 1998

Ono, Journal of Physical Chemistry C 112(), 2008
Energy-filtered transmission electron microscopy of biological samples on highly transparent carbon nanomembranes.
Rhinow D, Buenfeld M, Weber NE, Beyer A, Golzhauser A, Kuhlbrandt W, Hampp N, Turchanin A., Ultramicroscopy 111(5), 2011
PMID: 21329648
Contrast mechanisms and image formation in helium ion microscopy.
Bell DC., Microsc. Microanal. 15(2), 2009
PMID: 19284896
Design of a hybrid double-sideband/single-sideband (schlieren) objective aperture suitable for electron microscopy.
Buijsse B, van Laarhoven FM, Schmid AK, Cambie R, Cabrini S, Jin J, Glaeser RM., Ultramicroscopy 111(12), 2011
PMID: 22088443
Transmission electron microscopy at 20 kV for imaging and spectroscopy.
Kaiser U, Biskupek J, Meyer JC, Leschner J, Lechner L, Rose H, Stoger-Pollach M, Khlobystov AN, Hartel P, Muller H, Haider M, Eyhusen S, Benner G., Ultramicroscopy 111(8), 2011
PMID: 21801697
Selective sputtering and atomic resolution imaging of atomically thin boron nitride membranes.
Meyer JC, Chuvilin A, Algara-Siller G, Biskupek J, Kaiser U., Nano Lett. 9(7), 2009
PMID: 19480400
First application of Cc-corrected imaging for high-resolution and energy-filtered TEM.
Kabius B, Hartel P, Haider M, Muller H, Uhlemann S, Loebau U, Zach J, Rose H., J Electron Microsc (Tokyo) 58(3), 2009
PMID: 19398781
Optimizing the phase shift and the cut-on periodicity of phase plates for TEM.
Danev R, Nagayama K., Ultramicroscopy 111(8), 2011
PMID: 21864771


Markieren/ Markierung löschen
Markierte Publikationen

Open Data PUB

Web of Science

Dieser Datensatz im Web of Science®


PMID: 22537744
PubMed | Europe PMC

Suchen in

Google Scholar