Journal
ZEBRAFISH
Volume 2, Issue 2, Pages 105-111Publisher
MARY ANN LIEBERT, INC
DOI: 10.1089/zeb.2005.2.105
Keywords
-
Categories
Funding
- National Institutes of Health [R01 AI054503]
- Ellison Medical Foundation
- National Institute of General Medical Sciences [PHS NRSA T32 GM07270]
Ask authors/readers for more resources
One of the strengths of the zebrafish is the ease with which in situ hybridization can be performed to determine spatial and temporal patterns of gene expression in whole embryos. Thus far, colorimetric detection methods are mainly used for these analyses. Here we describe a fluorescent in situ hybridization (FISH) protocol for whole-mount zebrafish embryos using tyramide signal amplification (TSA). An optimal set of reagents was identified that allows for simultaneous localization of gene expression patterns of two genes within the same embryo, permitting identification of colocalized expression within single cells. This protocol can be extended to perform multiplex studies by repetition of the TSA-based detection for each target sequentially with a different fluorescent dye label. To this effect, we demonstrate that this approach can be combined with standard horseradish peroxidase (HRP)-mediated immunocytochemistry procedures in addition to FISH.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available