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Throughput and resolution with a next-generation direct electron detector

Direct electron detectors (DEDs) have revolutionized cryo-electron microscopy (cryo-EM) by facilitating the correction of beam-induced motion and radiation damage, and also by providing high-resolution image capture. A new-generation DED, the DE64, has been developed by Direct Electron that has good...

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Autores principales: Mendez, Joshua H., Mehrani, Atousa, Randolph, Peter, Stagg, Scott
Formato: Online Artículo Texto
Lenguaje:English
Publicado: International Union of Crystallography 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6830211/
https://www.ncbi.nlm.nih.gov/pubmed/31709056
http://dx.doi.org/10.1107/S2052252519012661
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author Mendez, Joshua H.
Mehrani, Atousa
Randolph, Peter
Stagg, Scott
author_facet Mendez, Joshua H.
Mehrani, Atousa
Randolph, Peter
Stagg, Scott
author_sort Mendez, Joshua H.
collection PubMed
description Direct electron detectors (DEDs) have revolutionized cryo-electron microscopy (cryo-EM) by facilitating the correction of beam-induced motion and radiation damage, and also by providing high-resolution image capture. A new-generation DED, the DE64, has been developed by Direct Electron that has good performance in both integrating and counting modes. The camera has been characterized in both modes in terms of image quality, throughput and resolution of cryo-EM reconstructions. The modulation transfer function, noise power spectrum and detective quantum efficiency (DQE) were determined for both modes, as well as the number of images per unit time. Although the DQE for counting mode was superior to that for integrating mode, the data-collection throughput for this mode was more than ten times slower. Since throughput and resolution are related in single-particle cryo-EM, data for apoferritin were collected and reconstructed using integrating mode, integrating mode in conjunction with a Volta phase plate (VPP) and counting mode. Only the counting-mode data resulted in a better than 3 Å resolution reconstruction with similar numbers of particles, and this increased performance could not be compensated for by the increased throughput of integrating mode or by the increased low-frequency contrast of integrating mode with the VPP. These data show that the superior image quality provided by counting mode is more important for high-resolution cryo-EM reconstructions than the superior throughput of integrating mode.
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spelling pubmed-68302112019-11-08 Throughput and resolution with a next-generation direct electron detector Mendez, Joshua H. Mehrani, Atousa Randolph, Peter Stagg, Scott IUCrJ Research Letters Direct electron detectors (DEDs) have revolutionized cryo-electron microscopy (cryo-EM) by facilitating the correction of beam-induced motion and radiation damage, and also by providing high-resolution image capture. A new-generation DED, the DE64, has been developed by Direct Electron that has good performance in both integrating and counting modes. The camera has been characterized in both modes in terms of image quality, throughput and resolution of cryo-EM reconstructions. The modulation transfer function, noise power spectrum and detective quantum efficiency (DQE) were determined for both modes, as well as the number of images per unit time. Although the DQE for counting mode was superior to that for integrating mode, the data-collection throughput for this mode was more than ten times slower. Since throughput and resolution are related in single-particle cryo-EM, data for apoferritin were collected and reconstructed using integrating mode, integrating mode in conjunction with a Volta phase plate (VPP) and counting mode. Only the counting-mode data resulted in a better than 3 Å resolution reconstruction with similar numbers of particles, and this increased performance could not be compensated for by the increased throughput of integrating mode or by the increased low-frequency contrast of integrating mode with the VPP. These data show that the superior image quality provided by counting mode is more important for high-resolution cryo-EM reconstructions than the superior throughput of integrating mode. International Union of Crystallography 2019-10-24 /pmc/articles/PMC6830211/ /pubmed/31709056 http://dx.doi.org/10.1107/S2052252519012661 Text en © Joshua H. Mendez et al. 2019 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 Letters
Mendez, Joshua H.
Mehrani, Atousa
Randolph, Peter
Stagg, Scott
Throughput and resolution with a next-generation direct electron detector
title Throughput and resolution with a next-generation direct electron detector
title_full Throughput and resolution with a next-generation direct electron detector
title_fullStr Throughput and resolution with a next-generation direct electron detector
title_full_unstemmed Throughput and resolution with a next-generation direct electron detector
title_short Throughput and resolution with a next-generation direct electron detector
title_sort throughput and resolution with a next-generation direct electron detector
topic Research Letters
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6830211/
https://www.ncbi.nlm.nih.gov/pubmed/31709056
http://dx.doi.org/10.1107/S2052252519012661
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