JoVE Logo

Sign In

Abstract

Biochemistry

Cryo-EM and Single-Particle Analysis with Scipion

Published: May 29th, 2021

DOI:

10.3791/62261

1Centro Nacional de Biotecnología, Campus Universidad Autónoma de Madrid, 2Faculty of Informatics, Masaryk University, 3Institute of Computer Science, Masaryk University, 4Campus Urbanización Montepríncipe, Universidad San Pablo CEU

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.

Explore More Videos

Keywords Cryo EM

This article has been published

Video Coming Soon

JoVE Logo

Privacy

Terms of Use

Policies

Research

Education

ABOUT JoVE

Copyright © 2024 MyJoVE Corporation. All rights reserved