PUBLICATION
Using single-molecule fluorescence in situ hybridization and immunohistochemistry to count RNA molecules in single cells in zebrafish embryos
- Authors
- Keseroglu, K., Zinani, O.Q.H., Özbudak, E.M.
- ID
- ZDB-PUB-230114-7
- Date
- 2023
- Source
- STAR protocols 4: 102020102020 (Journal)
- Registered Authors
- Keseroglu, Kemal
- Keywords
- Cell Biology, Developmental biology, In Situ Hybridization, Microscopy, Model Organisms, Molecular Biology, Molecular/Chemical Probes, Single-molecule Assays
- MeSH Terms
-
- Animals
- Embryonic Development*
- Female
- Immunohistochemistry
- In Situ Hybridization, Fluorescence
- RNA/genetics
- Zebrafish*/genetics
- PubMed
- 36638016 Full text @ STAR Protoc
Citation
Keseroglu, K., Zinani, O.Q.H., Özbudak, E.M. (2023) Using single-molecule fluorescence in situ hybridization and immunohistochemistry to count RNA molecules in single cells in zebrafish embryos. STAR protocols. 4:102020102020.
Abstract
Taming gene expression variability is critical for robust pattern formation during embryonic development. Here, we describe an optimized protocol for single-molecule fluorescence in situ hybridization and immunohistochemistry in zebrafish embryos. We detail how to count segmentation clock RNAs and calculate their variability among neighboring cells. This approach is easily adaptable to count RNA numbers of any gene and calculate transcriptional variability among neighboring cells in diverse biological settings. For complete details on the use and execution of this protocol, please refer to Keskin et al. (2018),1 Zinani et al. (2021),2 and Zinani et al. (2022).3.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping