Temporal dynamics of charge buildup in cryo-electron microscopy

Makoto Tokoro Schreiber, Alan Maigné, Marco Beleggia, Satoshi Shibata, Matthias Wolf*

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

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Abstract

It is well known that insulating samples can accumulate electric charges from exposure to an electron beam. How the accumulation of charge affects imaging parameters and sample stability in transmission electron microscopy is poorly understood. To quantify these effects, it is important to know how the charge is distributed within the sample and how it builds up over time. In the present study, we determine the spatial distribution and temporal dynamics of charge accumulation on vitreous ice samples with embedded proteins through a combination of modeling and Fresnel diffraction experiments. Our data reveal a rapid evolution of the charge state on ice upon initial exposure to the electron beam accompanied by charge gradients at the interfaces between ice and carbon films. We demonstrate that ice film movement and charge state variations occur upon electron beam exposure and are dose-rate dependent. Both affect the image defocus through a combination of sample height changes and lensing effects. Our results may be used as a guide to improve sample preparation, data collection, and data processing for imaging of dose-sensitive samples.
Original languageEnglish
Article number100081
JournalJournal of Structural Biology: X
Volume7
Number of pages9
ISSN2590-1524
DOIs
Publication statusPublished - 2023

Keywords

  • Cryo-electron microscopy
  • Specimen charging
  • Ice
  • Defocused diffraction
  • Fresnel diffraction

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