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Characterizing the resolution and throughput of the Apollo direct electron detector

Advances in electron detection have been essential to the success of high-resolution cryo-EM structure determination. A new generation of direct electron detector called the Apollo, has been developed by Direct Electron. The Apollo uses a novel event-based MAPS detector custom designed for ultra-fas...

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Autores principales: Peng, Ruizhi, Fu, Xiaofeng, Mendez, Joshua H., Randolph, Peter S., Bammes, Benjamin E., Stagg, Scott M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9791170/
https://www.ncbi.nlm.nih.gov/pubmed/36578473
http://dx.doi.org/10.1016/j.yjsbx.2022.100080
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author Peng, Ruizhi
Fu, Xiaofeng
Mendez, Joshua H.
Randolph, Peter S.
Bammes, Benjamin E.
Stagg, Scott M.
author_facet Peng, Ruizhi
Fu, Xiaofeng
Mendez, Joshua H.
Randolph, Peter S.
Bammes, Benjamin E.
Stagg, Scott M.
author_sort Peng, Ruizhi
collection PubMed
description Advances in electron detection have been essential to the success of high-resolution cryo-EM structure determination. A new generation of direct electron detector called the Apollo, has been developed by Direct Electron. The Apollo uses a novel event-based MAPS detector custom designed for ultra-fast electron counting. We have evaluated this new camera, finding that it delivers high detective quantum efficiency (DQE) and low coincidence loss, enabling high-quality electron counting data acquisition at up to nearly 80 input electrons per pixel per second. We further characterized the performance of Apollo for single particle cryo-EM on real biological samples. Using mouse apoferritin, Apollo yielded better than 1.9 Å resolution reconstructions at all three tested dose rates from a half-day data collection session each. With longer collection time and improved specimen preparation, mouse apoferritin was reconstructed to 1.66 Å resolution. Applied to a more challenging small protein aldolase, we obtained a 2.24 Å resolution reconstruction. The high quality of the map indicates that the Apollo has sufficiently high DQE to reconstruct smaller proteins and complexes with high-fidelity. Our results demonstrate that the Apollo camera performs well across a broad range of dose rates and is capable of capturing high quality data that produce high-resolution reconstructions for large and small single particle samples.
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spelling pubmed-97911702022-12-27 Characterizing the resolution and throughput of the Apollo direct electron detector Peng, Ruizhi Fu, Xiaofeng Mendez, Joshua H. Randolph, Peter S. Bammes, Benjamin E. Stagg, Scott M. J Struct Biol X Cryo-EM Methods: Resolving Details From Atoms to Cell Advances in electron detection have been essential to the success of high-resolution cryo-EM structure determination. A new generation of direct electron detector called the Apollo, has been developed by Direct Electron. The Apollo uses a novel event-based MAPS detector custom designed for ultra-fast electron counting. We have evaluated this new camera, finding that it delivers high detective quantum efficiency (DQE) and low coincidence loss, enabling high-quality electron counting data acquisition at up to nearly 80 input electrons per pixel per second. We further characterized the performance of Apollo for single particle cryo-EM on real biological samples. Using mouse apoferritin, Apollo yielded better than 1.9 Å resolution reconstructions at all three tested dose rates from a half-day data collection session each. With longer collection time and improved specimen preparation, mouse apoferritin was reconstructed to 1.66 Å resolution. Applied to a more challenging small protein aldolase, we obtained a 2.24 Å resolution reconstruction. The high quality of the map indicates that the Apollo has sufficiently high DQE to reconstruct smaller proteins and complexes with high-fidelity. Our results demonstrate that the Apollo camera performs well across a broad range of dose rates and is capable of capturing high quality data that produce high-resolution reconstructions for large and small single particle samples. Elsevier 2022-12-05 /pmc/articles/PMC9791170/ /pubmed/36578473 http://dx.doi.org/10.1016/j.yjsbx.2022.100080 Text en © 2022 The Author(s) https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Cryo-EM Methods: Resolving Details From Atoms to Cell
Peng, Ruizhi
Fu, Xiaofeng
Mendez, Joshua H.
Randolph, Peter S.
Bammes, Benjamin E.
Stagg, Scott M.
Characterizing the resolution and throughput of the Apollo direct electron detector
title Characterizing the resolution and throughput of the Apollo direct electron detector
title_full Characterizing the resolution and throughput of the Apollo direct electron detector
title_fullStr Characterizing the resolution and throughput of the Apollo direct electron detector
title_full_unstemmed Characterizing the resolution and throughput of the Apollo direct electron detector
title_short Characterizing the resolution and throughput of the Apollo direct electron detector
title_sort characterizing the resolution and throughput of the apollo direct electron detector
topic Cryo-EM Methods: Resolving Details From Atoms to Cell
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9791170/
https://www.ncbi.nlm.nih.gov/pubmed/36578473
http://dx.doi.org/10.1016/j.yjsbx.2022.100080
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