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Assessing the JEOL CRYO ARM 300 for high-throughput automated single-particle cryo-EM in a multiuser environment

Single-particle cryo-EM has become an indispensable structural biology method. It requires regular access to high-resolution electron cryogenic microscopes. To fully utilize the capacity of the expensive high-resolution instruments, the time used for data acquisition and the rate of data collection...

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Autores principales: Fislage, Marcus, Shkumatov, Alexander V., Stroobants, Annelore, Efremov, Rouslan G.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: International Union of Crystallography 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7340256/
https://www.ncbi.nlm.nih.gov/pubmed/32695417
http://dx.doi.org/10.1107/S2052252520006065
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author Fislage, Marcus
Shkumatov, Alexander V.
Stroobants, Annelore
Efremov, Rouslan G.
author_facet Fislage, Marcus
Shkumatov, Alexander V.
Stroobants, Annelore
Efremov, Rouslan G.
author_sort Fislage, Marcus
collection PubMed
description Single-particle cryo-EM has become an indispensable structural biology method. It requires regular access to high-resolution electron cryogenic microscopes. To fully utilize the capacity of the expensive high-resolution instruments, the time used for data acquisition and the rate of data collection have to be maximized. This in turn requires high stability and high uptime of the instrument. One of the first 300 kV JEOL CRYO ARM 300 microscopes has been installed at the cryo-EM facility BECM at VIB-VUB, Brussels, where the microscope is used for continuous data collection on multiple projects. Here, the suitability and performance of the microscope is assessed for high-throughput single-particle data collection. In particular, the properties of the illumination system, the stage stability and ice contamination rates are reported. The microscope was benchmarked using mouse heavy-chain apoferritin which was reconstructed to a resolution of 1.9 Å. Finally, uptime and throughput statistics of the instrument accumulated during the first six months of the facility operation in user access mode are reported.
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spelling pubmed-73402562020-07-20 Assessing the JEOL CRYO ARM 300 for high-throughput automated single-particle cryo-EM in a multiuser environment Fislage, Marcus Shkumatov, Alexander V. Stroobants, Annelore Efremov, Rouslan G. IUCrJ Research Papers Single-particle cryo-EM has become an indispensable structural biology method. It requires regular access to high-resolution electron cryogenic microscopes. To fully utilize the capacity of the expensive high-resolution instruments, the time used for data acquisition and the rate of data collection have to be maximized. This in turn requires high stability and high uptime of the instrument. One of the first 300 kV JEOL CRYO ARM 300 microscopes has been installed at the cryo-EM facility BECM at VIB-VUB, Brussels, where the microscope is used for continuous data collection on multiple projects. Here, the suitability and performance of the microscope is assessed for high-throughput single-particle data collection. In particular, the properties of the illumination system, the stage stability and ice contamination rates are reported. The microscope was benchmarked using mouse heavy-chain apoferritin which was reconstructed to a resolution of 1.9 Å. Finally, uptime and throughput statistics of the instrument accumulated during the first six months of the facility operation in user access mode are reported. International Union of Crystallography 2020-06-11 /pmc/articles/PMC7340256/ /pubmed/32695417 http://dx.doi.org/10.1107/S2052252520006065 Text en © Fislage 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
Fislage, Marcus
Shkumatov, Alexander V.
Stroobants, Annelore
Efremov, Rouslan G.
Assessing the JEOL CRYO ARM 300 for high-throughput automated single-particle cryo-EM in a multiuser environment
title Assessing the JEOL CRYO ARM 300 for high-throughput automated single-particle cryo-EM in a multiuser environment
title_full Assessing the JEOL CRYO ARM 300 for high-throughput automated single-particle cryo-EM in a multiuser environment
title_fullStr Assessing the JEOL CRYO ARM 300 for high-throughput automated single-particle cryo-EM in a multiuser environment
title_full_unstemmed Assessing the JEOL CRYO ARM 300 for high-throughput automated single-particle cryo-EM in a multiuser environment
title_short Assessing the JEOL CRYO ARM 300 for high-throughput automated single-particle cryo-EM in a multiuser environment
title_sort assessing the jeol cryo arm 300 for high-throughput automated single-particle cryo-em in a multiuser environment
topic Research Papers
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7340256/
https://www.ncbi.nlm.nih.gov/pubmed/32695417
http://dx.doi.org/10.1107/S2052252520006065
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