Protocol for Single-Molecule Fluorescence
Journal
STAR protocols
ISSN: 2666-1667
Titre abrégé: STAR Protoc
Pays: United States
ID NLM: 101769501
Informations de publication
Date de publication:
19 06 2020
19 06 2020
Historique:
entrez:
28
10
2020
pubmed:
29
10
2020
medline:
29
10
2020
Statut:
epublish
Résumé
We describe an optimized smFISH protocol for the intact pancreas. The protocol is adapted from Lyubimova et al. (2013), a generic tissue smFISH protocol that works for most tissues but not the pancreas. The main changes implemented include increasing the period of mRNA denaturation from 5 min to at least 3 h and increasing formamide concentrations from 10% to 30%. These modifications yield sensitive single mRNA visualization that is comparable to those achieved in other tissues using the standard protocol. For complete details on the use and execution of this protocol, please refer to Farack et al., 2018, Farack et al., 2019.
Identifiants
pubmed: 33111069
doi: 10.1016/j.xpro.2019.100007
pii: S2666-1667(19)30007-3
pmc: PMC7580104
doi:
Substances chimiques
RNA, Messenger
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Pagination
100007Subventions
Organisme : European Research Council
ID : 768956
Pays : International
Organisme : Howard Hughes Medical Institute
Pays : United States
Informations de copyright
© 2020.
Déclaration de conflit d'intérêts
The authors declare no competing interests.
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