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  • A protocol for whole-mount immuno-coupled hybridization chain reaction (WICHCR) in zebrafish embryos and larvae.

A protocol for whole-mount immuno-coupled hybridization chain reaction (WICHCR) in zebrafish embryos and larvae.

STAR protocols (2021-08-18)
Rodrigo Ibarra-García-Padilla, Aubrey Gaylon Adam Howard, Eileen Willey Singleton, Rosa Anna Uribe
ABSTRACT

Characterizing mRNA and protein expression with temporal and spatial resolution is a valuable component of nearly every developmental study. Here, we describe a protocol that combines in situ hybridization chain reaction (HCR) and immunofluorescence, allowing for the detection of mRNAs and proteins simultaneously, in zebrafish embryos and larvae. This protocol expands the flexibility of multiplexed HCR by coupling it with traditional immunofluorescence detection. For complete details on the use and execution of this protocol, please refer to Choi et al. (2010, 2016, 2018) and Howard et al. (2021).

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Paraformaldehyde, reagent grade, crystalline
Sigma-Aldrich
N-Phenylthiourea, ≥98%