Single Particle Cryo-Electron Microscopy: From Sample to Structure.
J Vis Exp
; (171)2021 05 29.
Article
en En
| MEDLINE
| ID: mdl-34125091
ABSTRACT
Cryo-electron microscopy (cryoEM) is a powerful technique for structure determination of macromolecular complexes, via single particle analysis (SPA). The overall process involves i) vitrifying the specimen in a thin film supported on a cryoEM grid; ii) screening the specimen to assess particle distribution and ice quality; iii) if the grid is suitable, collecting a single particle dataset for analysis; and iv) image processing to yield an EM density map. In this protocol, an overview for each of these steps is provided, with a focus on the variables which a user can modify during the workflow and the troubleshooting of common issues. With remote microscope operation becoming standard in many facilities, variations on imaging protocols to assist users in efficient operation and imaging when physical access to the microscope is limited will be described.
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Procesamiento de Imagen Asistido por Computador
Idioma:
En
Revista:
J Vis Exp
Año:
2021
Tipo del documento:
Article