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Charging of Vitreous Samples in Cryogenic Electron Microscopy Mitigated by Graphene
[Image: see text] Cryogenic electron microscopy can provide high-resolution reconstructions of macromolecules embedded in a thin layer of ice from which atomic models can be built de novo. However, the interaction between the ionizing electron beam and the sample results in beam-induced motion and i...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Chemical Society
2023
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10448747/ https://www.ncbi.nlm.nih.gov/pubmed/37531407 http://dx.doi.org/10.1021/acsnano.3c03722 |
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author | Zhang, Yue van Schayck, J. Paul Pedrazo-Tardajos, Adrián Claes, Nathalie Noteborn, Willem E. M. Lu, Peng-Han Duimel, Hans Dunin-Borkowski, Rafal E. Bals, Sara Peters, Peter J. Ravelli, Raimond B. G. |
author_facet | Zhang, Yue van Schayck, J. Paul Pedrazo-Tardajos, Adrián Claes, Nathalie Noteborn, Willem E. M. Lu, Peng-Han Duimel, Hans Dunin-Borkowski, Rafal E. Bals, Sara Peters, Peter J. Ravelli, Raimond B. G. |
author_sort | Zhang, Yue |
collection | PubMed |
description | [Image: see text] Cryogenic electron microscopy can provide high-resolution reconstructions of macromolecules embedded in a thin layer of ice from which atomic models can be built de novo. However, the interaction between the ionizing electron beam and the sample results in beam-induced motion and image distortion, which limit the attainable resolutions. Sample charging is one contributing factor of beam-induced motions and image distortions, which is normally alleviated by including part of the supporting conducting film within the beam-exposed region. However, routine data collection schemes avoid strategies whereby the beam is not in contact with the supporting film, whose rationale is not fully understood. Here we characterize electrostatic charging of vitreous samples, both in imaging and in diffraction mode. We mitigate sample charging by depositing a single layer of conductive graphene on top of regular EM grids. We obtained high-resolution single-particle analysis (SPA) reconstructions at 2 Å when the electron beam only irradiates the middle of the hole on graphene-coated grids, using data collection schemes that previously failed to produce sub 3 Å reconstructions without the graphene layer. We also observe that the SPA data obtained with the graphene-coated grids exhibit a higher b factor and reduced particle movement compared to data obtained without the graphene layer. This mitigation of charging could have broad implications for various EM techniques, including SPA and cryotomography, and for the study of radiation damage and the development of future sample carriers. Furthermore, it may facilitate the exploration of more dose-efficient, scanning transmission EM based SPA techniques. |
format | Online Article Text |
id | pubmed-10448747 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | American Chemical Society |
record_format | MEDLINE/PubMed |
spelling | pubmed-104487472023-08-25 Charging of Vitreous Samples in Cryogenic Electron Microscopy Mitigated by Graphene Zhang, Yue van Schayck, J. Paul Pedrazo-Tardajos, Adrián Claes, Nathalie Noteborn, Willem E. M. Lu, Peng-Han Duimel, Hans Dunin-Borkowski, Rafal E. Bals, Sara Peters, Peter J. Ravelli, Raimond B. G. ACS Nano [Image: see text] Cryogenic electron microscopy can provide high-resolution reconstructions of macromolecules embedded in a thin layer of ice from which atomic models can be built de novo. However, the interaction between the ionizing electron beam and the sample results in beam-induced motion and image distortion, which limit the attainable resolutions. Sample charging is one contributing factor of beam-induced motions and image distortions, which is normally alleviated by including part of the supporting conducting film within the beam-exposed region. However, routine data collection schemes avoid strategies whereby the beam is not in contact with the supporting film, whose rationale is not fully understood. Here we characterize electrostatic charging of vitreous samples, both in imaging and in diffraction mode. We mitigate sample charging by depositing a single layer of conductive graphene on top of regular EM grids. We obtained high-resolution single-particle analysis (SPA) reconstructions at 2 Å when the electron beam only irradiates the middle of the hole on graphene-coated grids, using data collection schemes that previously failed to produce sub 3 Å reconstructions without the graphene layer. We also observe that the SPA data obtained with the graphene-coated grids exhibit a higher b factor and reduced particle movement compared to data obtained without the graphene layer. This mitigation of charging could have broad implications for various EM techniques, including SPA and cryotomography, and for the study of radiation damage and the development of future sample carriers. Furthermore, it may facilitate the exploration of more dose-efficient, scanning transmission EM based SPA techniques. American Chemical Society 2023-08-02 /pmc/articles/PMC10448747/ /pubmed/37531407 http://dx.doi.org/10.1021/acsnano.3c03722 Text en © 2023 The Authors. Published by American Chemical Society https://creativecommons.org/licenses/by/4.0/Permits the broadest form of re-use including for commercial purposes, provided that author attribution and integrity are maintained (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Zhang, Yue van Schayck, J. Paul Pedrazo-Tardajos, Adrián Claes, Nathalie Noteborn, Willem E. M. Lu, Peng-Han Duimel, Hans Dunin-Borkowski, Rafal E. Bals, Sara Peters, Peter J. Ravelli, Raimond B. G. Charging of Vitreous Samples in Cryogenic Electron Microscopy Mitigated by Graphene |
title | Charging
of Vitreous Samples in Cryogenic Electron
Microscopy Mitigated by Graphene |
title_full | Charging
of Vitreous Samples in Cryogenic Electron
Microscopy Mitigated by Graphene |
title_fullStr | Charging
of Vitreous Samples in Cryogenic Electron
Microscopy Mitigated by Graphene |
title_full_unstemmed | Charging
of Vitreous Samples in Cryogenic Electron
Microscopy Mitigated by Graphene |
title_short | Charging
of Vitreous Samples in Cryogenic Electron
Microscopy Mitigated by Graphene |
title_sort | charging
of vitreous samples in cryogenic electron
microscopy mitigated by graphene |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10448747/ https://www.ncbi.nlm.nih.gov/pubmed/37531407 http://dx.doi.org/10.1021/acsnano.3c03722 |
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