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Analyzing contrast in cryo-transmission electron microscopy: Comparison of electrostatic Zach phase plates and hole-free phase plates

Obermair, Martin; Hettler, Simon; Hsieh, Chyongere; Dries, Manuel; Marko, Michael; Gerthsen, Dagmar

Abstract:
Phase plates (PPs) are beneficial devices to improve the phase contrast of life-science objects in cryo-transmission electron microscopy (TEM). The development of the hole-free (HF) PP, which consists of a thin carbon film, has led to impressive results due to its ease in fabrication, implementation and application. However, the phase shift of the HFPP can be controlled only indirectly. The electrostatic Zach PP uses a strongly localized and adjustable electrostatic potential to generate well-defined and variable phase shifts between scattered and unscattered electrons. However, artifacts in phase-contrast TEM images are induced by the presence of the PP rod in the diffraction plane. We present a detailed analysis and comparison of the contrast-enhancing capabilities of both PP types and their emerging artifacts. For this purpose, cryo-TEM images of a standard T4-bacteriophage test sample were acquired with both PP types. Simulated images reproduce the experimental images well and substantially contribute to the understanding of contrast formation. An electrostatic Zach PP was used in this work to acquire cryo-electron tomograms with enhanced contrast, which are of similar quality as tomograms obtained by HFPP TEM.

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Verlagsausgabe §
DOI: 10.5445/IR/1000129902
Veröffentlicht am 19.02.2021
Cover der Publikation
Zugehörige Institution(en) am KIT Laboratorium für Elektronenmikroskopie (LEM)
Publikationstyp Zeitschriftenaufsatz
Publikationsmonat/-jahr 11.2020
Sprache Englisch
Identifikator ISSN: 0304-3991
KITopen-ID: 1000129902
Erschienen in Ultramicroscopy
Verlag Elsevier
Band 218
Seiten Art.-Nr.: 113086
Schlagwörter Cryo-transmission electron microscopy (TEM); Phase plate; Phase contrast; Thin film; Biological specimen; Cryo-electron tomography (cryo-ET)
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