Significant regulation of gene expression is mediated at the translation level. Here, we describe
protocols for imaging and analysis of translation at single mRNA resolution in both fixed and living
Drosophila embryos. These protocols use the SunTag system, in which the protein of interest is
visualized by inserting a peptide array that is recognized by a single chain antibody. Simultaneous
detection of individual mRNAs using the MS2/MCP system or by smFISH allows translation sites to be
identified and quantified.