A protocol for cryogenic volumetric imaging using serial plasma FIB/SEM.
Methods Cell Biol
; 177: 327-358, 2023.
Article
em En
| MEDLINE
| ID: mdl-37451772
ABSTRACT
Cryogenic volumetric imaging using serial plasma focused ion beam scanning electron microscopy (serial pFIB/SEM) is a new and exciting correlative volume electron microscopy (vEM) technique. It enables visualization of un-stained, cryogenically immobilized cells and tissues with â¼20-50nm resolution and a field of view of â¼10-30µm resulting in near-native state imaging and the possibility of microscale, mesoscale and nanoscale correlative imaging. We have written a detailed protocol for optimization of FIB and SEM parameters to reduce imaging artefacts and enable downstream computational processing and analysis. While our experience is based on use of a single system, the protocol has been written to be as hardware and software agnostic as possible, with a focus on the purpose of each step rather than a fully procedural description to provide a useful resource regardless of the system/software in use.
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Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Imageamento Tridimensional
/
Microscopia Eletrônica de Volume
Idioma:
En
Ano de publicação:
2023
Tipo de documento:
Article