Temporal dynamics of charge buildup in cryo-electron microscopy
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...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-01-01
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| Series: | Journal of Structural Biology: X |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2590152422000228 |
| _version_ | 1797798661693898752 |
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| author | Makoto Tokoro Schreiber Alan Maigné Marco Beleggia Satoshi Shibata Matthias Wolf |
| author_facet | Makoto Tokoro Schreiber Alan Maigné Marco Beleggia Satoshi Shibata Matthias Wolf |
| author_sort | Makoto Tokoro Schreiber |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-03-13T04:07:12Z |
| format | Article |
| id | doaj.art-4d50ea974de64042b04fc8f8c21cbbab |
| institution | Directory Open Access Journal |
| issn | 2590-1524 |
| language | English |
| last_indexed | 2024-03-13T04:07:12Z |
| publishDate | 2023-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Structural Biology: X |
| spelling | doaj.art-4d50ea974de64042b04fc8f8c21cbbab2023-06-21T06:59:05ZengElsevierJournal of Structural Biology: X2590-15242023-01-017100081Temporal dynamics of charge buildup in cryo-electron microscopyMakoto Tokoro Schreiber0Alan Maigné1Marco Beleggia2Satoshi Shibata3Matthias Wolf4Molecular Cryo-Electron Microscopy Unit, Okinawa Institute of Science and Technology Graduate University, 1919-1 Tancha, Onna-son, Okinawa, JapanMolecular Cryo-Electron Microscopy Unit, Okinawa Institute of Science and Technology Graduate University, 1919-1 Tancha, Onna-son, Okinawa, JapanDTU Nanolab, Technical University of Denmark, 2800 Kgs. Lyngby, DenmarkMolecular Cryo-Electron Microscopy Unit, Okinawa Institute of Science and Technology Graduate University, 1919-1 Tancha, Onna-son, Okinawa, JapanMolecular Cryo-Electron Microscopy Unit, Okinawa Institute of Science and Technology Graduate University, 1919-1 Tancha, Onna-son, Okinawa, Japan; Institute of Biological Chemistry, Academia Sinica, Nankang, Taipei 115, Taiwan; Corresponding author at: Molecular Cryo-Electron Microscopy Unit, Okinawa Institute of Science and Technology Graduate University, 1919-1 Tancha, Onna-son, Okinawa, Japan.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.http://www.sciencedirect.com/science/article/pii/S2590152422000228Cryo-electron microscopySpecimen chargingIceDefocused diffractionFresnel diffraction |
| spellingShingle | Makoto Tokoro Schreiber Alan Maigné Marco Beleggia Satoshi Shibata Matthias Wolf Temporal dynamics of charge buildup in cryo-electron microscopy Journal of Structural Biology: X Cryo-electron microscopy Specimen charging Ice Defocused diffraction Fresnel diffraction |
| title | Temporal dynamics of charge buildup in cryo-electron microscopy |
| title_full | Temporal dynamics of charge buildup in cryo-electron microscopy |
| title_fullStr | Temporal dynamics of charge buildup in cryo-electron microscopy |
| title_full_unstemmed | Temporal dynamics of charge buildup in cryo-electron microscopy |
| title_short | Temporal dynamics of charge buildup in cryo-electron microscopy |
| title_sort | temporal dynamics of charge buildup in cryo electron microscopy |
| topic | Cryo-electron microscopy Specimen charging Ice Defocused diffraction Fresnel diffraction |
| url | http://www.sciencedirect.com/science/article/pii/S2590152422000228 |
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