Assembly and Imaging set up of PIE-Scope.
Bio Protoc
; 10(19): e3768, 2020 Oct 05.
Article
en En
| MEDLINE
| ID: mdl-33659426
ABSTRACT
Cryo-Electron Tomography (cryo-ET) is a method that enables resolving the structure of macromolecular complexes directly in the cellular environment. However, sample preparation for in situ Cryo-ET is labour-intensive and can require both cryo-lamella preparation through cryo-Focused Ion Beam (FIB) milling and correlative light microscopy to ensure that the event of interest is present in the lamella. Here, we present an integrated cryo-FIB and light microscope setup called the Photon Ion Electron microscope (PIE-scope) that enables direct and rapid isolation of cellular regions containing protein complexes of interest. The PIE-scope can be retrofitted on existing microscopes, although the drawings we provide are meant to work on ThermoFisher DualBeams with small mechanical modifications those can be adapted on other brands.
Texto completo:
1
Bases de datos:
MEDLINE
Idioma:
En
Revista:
Bio Protoc
Año:
2020
Tipo del documento:
Article
País de afiliación:
Australia