CLEMSite, a software for automated phenotypic screens using light microscopy and FIB-SEM.
J Cell Biol
; 222(3)2023 03 06.
Article
in En
| MEDLINE
| ID: mdl-36562752
ABSTRACT
In recent years, Focused Ion Beam Scanning Electron Microscopy (FIB-SEM) has emerged as a flexible method that enables semi-automated volume ultrastructural imaging. We present a toolset for adherent cells that enables tracking and finding cells, previously identified in light microscopy (LM), in the FIB-SEM, along with the automatic acquisition of high-resolution volume datasets. We detect the underlying grid pattern in both modalities (LM and EM), to identify common reference points. A combination of computer vision techniques enables complete automation of the workflow. This includes setting the coincidence point of both ion and electron beams, automated evaluation of the image quality and constantly tracking the sample position with the microscope's field of view reducing or even eliminating operator supervision. We show the ability to target the regions of interest in EM within 5 µm accuracy while iterating between different targets and implementing unattended data acquisition. Our results demonstrate that executing volume acquisition in multiple locations autonomously is possible in EM.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Imaging, Three-Dimensional
/
Volume Electron Microscopy
Type of study:
Prognostic_studies
Language:
En
Journal:
J Cell Biol
Year:
2023
Document type:
Article
Affiliation country:
Germany