Quantification of tissue-specific protein translation in whole using O-propargyl-puromycin labeling and fluorescence microscopy.

The regulation of gene expression via protein translation is critical for growth, development, and stress response. While puromycin-based techniques have been used to quantify protein translation in , they have been limited to using lysate from whole worms. To achieve tissue-specific quantification of ribosome activity in intact , we report the application of O-propargyl-puromycin in a cuticle defective mutant followed by conjugation of an azide fluorophore for detection using fluorescent confocal microscopy.