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Fast and accurate defocus modulation for improved tunability of cryo-EM experiments
Current data collection strategies in electron cryo-microscopy (cryo-EM) record multiframe movies with static optical settings. This limits the number of adjustable parameters that can be used to optimize the experiment. Here, a method for fast and accurate defocus (FADE) modulation during movie acq...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
International Union of Crystallography
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7201282/ https://www.ncbi.nlm.nih.gov/pubmed/32431839 http://dx.doi.org/10.1107/S205225252000408X |
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author | Danev, Radostin Iijima, Hirofumi Matsuzaki, Mizuki Motoki, Sohei |
author_facet | Danev, Radostin Iijima, Hirofumi Matsuzaki, Mizuki Motoki, Sohei |
author_sort | Danev, Radostin |
collection | PubMed |
description | Current data collection strategies in electron cryo-microscopy (cryo-EM) record multiframe movies with static optical settings. This limits the number of adjustable parameters that can be used to optimize the experiment. Here, a method for fast and accurate defocus (FADE) modulation during movie acquisition is proposed. It uses the objective lens aperture as an electrostatic pole that locally modifies the electron beam potential. The beam potential variation is converted to defocus change by the typically undesired chromatic aberration of the objective lens. The simplicity, electrostatic principle and low electrical impedance of the device allow fast switching speeds that will enable per-frame defocus modulation of cryo-EM movies. Researchers will be able to define custom defocus ‘recipes’ and tailor the experiment for optimal information extraction from the sample. The FADE method could help to convert the microscope into a more dynamic and flexible optical platform that delivers better performance in cryo-EM single-particle analysis and electron cryo-tomography. |
format | Online Article Text |
id | pubmed-7201282 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | International Union of Crystallography |
record_format | MEDLINE/PubMed |
spelling | pubmed-72012822020-05-19 Fast and accurate defocus modulation for improved tunability of cryo-EM experiments Danev, Radostin Iijima, Hirofumi Matsuzaki, Mizuki Motoki, Sohei IUCrJ Research Papers Current data collection strategies in electron cryo-microscopy (cryo-EM) record multiframe movies with static optical settings. This limits the number of adjustable parameters that can be used to optimize the experiment. Here, a method for fast and accurate defocus (FADE) modulation during movie acquisition is proposed. It uses the objective lens aperture as an electrostatic pole that locally modifies the electron beam potential. The beam potential variation is converted to defocus change by the typically undesired chromatic aberration of the objective lens. The simplicity, electrostatic principle and low electrical impedance of the device allow fast switching speeds that will enable per-frame defocus modulation of cryo-EM movies. Researchers will be able to define custom defocus ‘recipes’ and tailor the experiment for optimal information extraction from the sample. The FADE method could help to convert the microscope into a more dynamic and flexible optical platform that delivers better performance in cryo-EM single-particle analysis and electron cryo-tomography. International Union of Crystallography 2020-04-25 /pmc/articles/PMC7201282/ /pubmed/32431839 http://dx.doi.org/10.1107/S205225252000408X Text en © Danev et al. 2020 http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution (CC-BY) Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are cited.http://creativecommons.org/licenses/by/4.0/ |
spellingShingle | Research Papers Danev, Radostin Iijima, Hirofumi Matsuzaki, Mizuki Motoki, Sohei Fast and accurate defocus modulation for improved tunability of cryo-EM experiments |
title | Fast and accurate defocus modulation for improved tunability of cryo-EM experiments |
title_full | Fast and accurate defocus modulation for improved tunability of cryo-EM experiments |
title_fullStr | Fast and accurate defocus modulation for improved tunability of cryo-EM experiments |
title_full_unstemmed | Fast and accurate defocus modulation for improved tunability of cryo-EM experiments |
title_short | Fast and accurate defocus modulation for improved tunability of cryo-EM experiments |
title_sort | fast and accurate defocus modulation for improved tunability of cryo-em experiments |
topic | Research Papers |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7201282/ https://www.ncbi.nlm.nih.gov/pubmed/32431839 http://dx.doi.org/10.1107/S205225252000408X |
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