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In-Focus Electrostatic Zach Phase Plate Imaging for Transmission Electron Microscopy with Tunable Phase Contrast of Frozen Hydrated Biological Samples

Published online by Cambridge University Press:  02 January 2014

Nicole Frindt ,
Marco Oster ,
Simon Hettler ,
Björn Gamm ,
Levin Dieterle ,
Wolfgang Kowalsky ,
Dagmar Gerthsen  and
Rasmus R. Schröder
Nicole Frindt*
Affiliation:
CellNetworks, BioQuant, Universitätsklinikum Heidelberg, Im Neuenheimer Feld 267, 69120 Heidelberg, Germany
Marco Oster
Affiliation:
CellNetworks, BioQuant, Universitätsklinikum Heidelberg, Im Neuenheimer Feld 267, 69120 Heidelberg, Germany
Simon Hettler
Affiliation:
Laboratorium für Elektronenmikroskopie, Karlsruhe Institut für Technologie (KIT), Engesserstr 7, 76128 Karlsruhe, Germany
Björn Gamm
Affiliation:
Laboratorium für Elektronenmikroskopie, Karlsruhe Institut für Technologie (KIT), Engesserstr 7, 76128 Karlsruhe, Germany
Levin Dieterle
Affiliation:
Institut für Hochfrequenztechnik, Technische Universität Braunschweig, Schleinitzstr 22, 38106 Braunschweig, Germany
Wolfgang Kowalsky
Affiliation:
Institut für Hochfrequenztechnik, Technische Universität Braunschweig, Schleinitzstr 22, 38106 Braunschweig, Germany Innovation Lab GmbH, Heidelberg, Speyerer Str. 4, 69115 Heidelberg, Germany
Dagmar Gerthsen
Affiliation:
Laboratorium für Elektronenmikroskopie, Karlsruhe Institut für Technologie (KIT), Engesserstr 7, 76128 Karlsruhe, Germany
Rasmus R. Schröder*
Affiliation:
CellNetworks, BioQuant, Universitätsklinikum Heidelberg, Im Neuenheimer Feld 267, 69120 Heidelberg, Germany Innovation Lab GmbH, Heidelberg, Speyerer Str. 4, 69115 Heidelberg, Germany CAM Centre of Advanced Materials, Universität Heidelberg, Im Neuenheimer Feld 227, 69120 Heidelberg, Germany
*
*Corresponding author. E-mail: nicole.frindt@googlemail.com
**Corresponding author. E-mail: rasmus.schroeder@bioquant.uni-heidelberg.de
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Abstract

Transmission electron microscopy (TEM) images of beam sensitive weak-phase objects such as biological cryo samples usually show a very low signal-to-noise ratio. These samples have almost no amplitude contrast and instead structural information is mainly encoded in the phase contrast. To increase the sample contrast in the image, especially for low spatial frequencies, the use of phase plates for close to focus phase contrast enhancement in TEM has long been discussed. Electrostatic phase plates are favorable in particular, as their tunable potential will allow an optimal phase shift adjustment and higher resolution than film phase plates as they avoid additional scattering events in matter. Here we show the first realization of close to focus phase contrast images of actin filament cryo samples acquired using an electrostatic Zach phase plate. Both positive and negative phase contrast is shown, which is obtained by applying appropriate potentials to the phase plate. The dependence of phase contrast improvement on sample orientation with respect to the phase plate is demonstrated and single-sideband artifacts are discussed. Additionally, possibilities to reduce contamination and charging effects of the phase plate are shown.

Keywords

electrostatic phase platecryo transmission electron microscopyweak-phase objectphase contrastfrozen hydrated biological samplescryo samplessingle-sideband effectchargingcontamination
Type
Biological Applications
Information
Microscopy and Microanalysis , Volume 20 , Issue 1 , February 2014 , pp. 175 - 183
Copyright
Copyright © Microscopy Society of America 2014 

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