Quality assessment in light microscopy for routine use through simple tools and robust metrics.
Journal
The Journal of cell biology
ISSN: 1540-8140
Titre abrégé: J Cell Biol
Pays: United States
ID NLM: 0375356
Informations de publication
Date de publication:
07 11 2022
07 11 2022
Historique:
received:
15
07
2021
revised:
04
04
2022
accepted:
31
08
2022
entrez:
29
9
2022
pubmed:
30
9
2022
medline:
4
10
2022
Statut:
ppublish
Résumé
Although there is a need to demonstrate reproducibility in light microscopy acquisitions, the lack of standardized guidelines monitoring microscope health status over time has so far impaired the widespread use of quality control (QC) measurements. As scientists from 10 imaging core facilities who encounter various types of projects, we provide affordable hardware and open source software tools, rigorous protocols, and define reference values to assess QC metrics for the most common fluorescence light microscopy modalities. Seven protocols specify metrics on the microscope resolution, field illumination flatness, chromatic aberrations, illumination power stability, stage drift, positioning repeatability, and spatial-temporal noise of camera sensors. We designed the MetroloJ_QC ImageJ/Fiji Java plugin to incorporate the metrics and automate analysis. Measurements allow us to propose an extensive characterization of the QC procedures that can be used by any seasoned microscope user, from research biologists with a specialized interest in fluorescence light microscopy through to core facility staff, to ensure reproducible and quantifiable microscopy results.
Identifiants
pubmed: 36173380
pii: 213512
doi: 10.1083/jcb.202107093
pmc: PMC9526251
pii:
doi:
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Subventions
Organisme : French National Research Agency
ID : ANR-10-INBS-04
Organisme : Centre National de la Recherche Scientifique
Informations de copyright
© 2022 Faklaris et al.
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