Cryo-EM and Single-Particle Analysis with Scipion
| Autor: | Yunior C. Fonseca, José María Carazo, E. Fernández-Giménez, Erney Ramírez-Aportela, Roberto Marabini, Pablo Conesa, Patricia Losana, L. del Cano, David Strelak, Jose Ramon Macias, David Maluenda, Ana Maria Cuervo, F. de Isidro-Gomez, Carlos Oscar S. Sorzano, María Jesús Martínez, A. Jiménez-Moreno, Rubén J. Sánchez-García, J. Jiménez de la Morena, David Herreros, Remedios Melero |
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| Přispěvatelé: | UAM. Departamento de Ingeniería Informática |
| Rok vydání: | 2021 |
| Předmět: |
Microscope
Traceability Macromolecular Substances Computer science General Chemical Engineering ComputingMethodologies_IMAGEPROCESSINGANDCOMPUTERVISION Single particle analysis Image processing Scipion 010402 general chemistry 01 natural sciences Column (database) macromolecul near-atomic resolution General Biochemistry Genetics and Molecular Biology Workflow law.invention law Computer graphics (images) Image Processing Computer-Assisted Cryo-electron microscop Informática General Immunology and Microbiology 010405 organic chemistry General Neuroscience Cryoelectron Microscopy 3D reconstruction Sample (graphics) Single Molecule Imaging 0104 chemical sciences |
| Zdroj: | Biblos-e Archivo. Repositorio Institucional de la UAM instname Journal of Visualized Experiments |
| ISSN: | 1940-087X |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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