High-Throughput DNA FISH (hiFISH).
3D genome
DNA FISH
High-throughput imaging
Nuclear architecture
Journal
Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
Titre abrégé: Methods Mol Biol
Pays: United States
ID NLM: 9214969
Informations de publication
Date de publication:
2022
2022
Historique:
entrez:
22
7
2022
pubmed:
23
7
2022
medline:
27
7
2022
Statut:
ppublish
Résumé
High-throughput DNA fluorescence in situ hybridization (hiFISH) combines multicolor combinatorial DNA FISH staining with automated image acquisition and analysis to visualize and localize tens to hundreds of genomic loci in up to millions of cells. hiFISH can be used to measure physical distances between pairs of genomic loci, radial distances from genomic loci to the nuclear edge or center, and distances between genomic loci and nuclear structures defined by protein or RNA markers. The resulting large datasets of 3D spatial distances can be used to study cellular heterogeneity in genome architecture and the molecular mechanisms underlying this phenomenon in a variety of cellular systems. In this chapter we provide detailed protocols for hiFISH to measure distances between genomic loci, including all steps involved in DNA FISH probe design and preparation, cell culture, DNA FISH staining in 384-well imaging plates, automated image acquisition and analysis, and, finally, statistical analysis.
Identifiants
pubmed: 35867253
doi: 10.1007/978-1-0716-2497-5_12
pmc: PMC10372930
mid: NIHMS1916268
doi:
Substances chimiques
DNA Probes
0
DNA
9007-49-2
Types de publication
Journal Article
Research Support, N.I.H., Intramural
Langues
eng
Sous-ensembles de citation
IM
Pagination
245-274Subventions
Organisme : Intramural NIH HHS
ID : Z01 BC010309
Pays : United States
Organisme : Intramural NIH HHS
ID : ZIA BC010309
Pays : United States
Organisme : Intramural NIH HHS
ID : ZIC BC011567
Pays : United States
Informations de copyright
© 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.
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