Zebrafish Whole Mount High-Resolution Double Fluorescent In Situ Hybridization

This procedure allows us to determine patterns of gene expression in zebrafish embryos at high resolution, an antisense ribo probe is hybridized to the complementary target mRNA. The labeled Ribo probe is recognized by an antibody that is conjugated to a peroxidase enzyme aramide conjugated fluoro four is added to the embryos and the peroxidase converts the IDE to a highly reactive compound. Leading to stable deposition of the fluoro four unbound IDE is washed away revealing the detailed subcellular pattern of mRNA distribution.

Expression of a second gene can be observed using a distinctly labeled probe and a different tyrom fluoro four, allowing the detailed comparison of expression. Hi, I'm Tim Brent from the laboratory of Scott Holly in the Department of Molecular, cellular and Developmental Biology at Yale University. Today we will show you a procedure for whole mount high resolution double fluorescent in situ two hybridization of zebra zebrafish embryos in all laboratory.