Methods for Double Detection of Gene Expression: Combined In Situ Hybridization and Immunocytochemistry or Histochemistry

The distribution of two different molecules can be analyzed within the same embryo using the procedures described below. The protocol for combined whole mount in situ hybridization and immunocytochemistry allows for simultaneous detection of messenger ribonucleic acid (mRNA) and protein. The protocol for combined whole mount in situ hybridization and β-galactosidase staining allows for simultaneous detection of mRNA and transgene-directed β-galactosidase expression. Simultaneous detection allows for the most direct comparison of expression patterns. These procedures are derived from protocols used in Drosophila (1 ) and mice (2 ,3 ).