Cryo-EM and single-particle analysis with Scipion
Author
Jiménez-Moreno, A.; Del Caño, L.; Martínez, M.; Ramírez-Aportela, E.; Cuervo, A.; Melero, R.; Sánchez-García, R.; Strelak, D.; Fernández-Giménez, E.; de Isidro-Gómez, F. P.; Herreros, D.; Conesa, P.; Fonseca, Y.; Maluenda, D.; Jiménez de la Morena, J.; Macías, J. R.; Losana, P.; Marabini Ruiz, Roberto; Carazo, J. M.; Sorzano, C. O.S.Entity
UAM. Departamento de Ingeniería InformáticaPublisher
MYJoVE CorporationDate
2021-05-29Citation
10.3791/62261
Journal of Visualized Experiments. JoVE 171 (2021): e62261
ISSN
1940-087XDOI
10.3791/62261Editor's Version
https://doi.org/10.3791/62261.Subjects
Cryo-electron microscop; macromolecul; near-atomic resolution; Scipion; InformáticaRights
© 2021 JoVEEsta obra está bajo una licencia de Creative Commons Reconocimiento-NoComercial-SinObraDerivada 4.0 Internacional.
Abstract
Cryo-electron microscopy has become one of the most important tools in biological research to reveal the structural information of macromolecules at near-atomic resolution. In single-particle analysis, the vitrified sample is imaged by an electron beam and the detectors at the end of the microscope column produce movies of that sample. These movies contain thousands of images of identical particles in random orientations. The data need to go through an image processing workflow with multiple steps to obtain the final 3D reconstructed volume. The goal of the image processing workflow is to identify the acquisition parameters to be able to reconstruct the specimen under study. Scipion provides all the tools to create this workflow using several image processing packages in an integrative framework, also allowing the traceability of the results. In this article the whole image processing workflow in Scipion is presented and discussed with data coming from a real test case, giving all the details necessary to go from the movies obtained by the microscope to a high resolution final 3D reconstruction. Also, the power of using consensus tools that allow combining methods, and confirming results along every step of the workflow, improving the accuracy of the obtained results, is discussed.
Files in this item
Google Scholar:Jiménez-Moreno, A.
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Del Caño, L.
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Martínez, M.
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Ramírez-Aportela, E.
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Cuervo, A.
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Melero, R.
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Sánchez-García, R.
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Strelak, D.
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Fernández-Giménez, E.
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de Isidro-Gómez, F. P.
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Herreros, D.
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Conesa, P.
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Fonseca, Y.
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Maluenda, D.
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Jiménez de la Morena, J.
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Macías, J. R.
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Losana, P.
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Marabini Ruiz, Roberto
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Carazo, J. M.
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Sorzano, C. O.S.
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