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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...

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Autores principales: Danev, Radostin, Iijima, Hirofumi, Matsuzaki, Mizuki, Motoki, Sohei
Formato: Online Artículo Texto
Lenguaje:English
Publicado: International Union of Crystallography 2020
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.
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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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