Cryo-EM and single-particle analysis with Scipion

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...

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Bibliographic Details
Authors: 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.
Format: article
Publication Date:2021
Country:España
Institution:Universidad Autónoma de Madrid
Repository:Biblos-e Archivo. Repositorio Institucional de la UAM
Language:English
OAI Identifier:oai:repositorio.uam.es:10486/697520
Online Access:http://hdl.handle.net/10486/697520
https://dx.doi.org/10.3791/62261
Access Level:Open access
Keyword:Cryo-electron microscop
macromolecul
near-atomic resolution
Scipion
Informática
Description
Summary: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.

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