Protocol for high-sensitivity detection of microRNAs in adult Drosophila tissues using hybridization chain reaction

MicroRNAs (miRNAs) are post-transcriptional regulators of gene expression and cell-fate determination. Here, we present a protocol for detecting miRNAs in Drosophila gonadal tissues using an optimized hybridization chain reaction (HCR)-based approach. We describe steps for using modular probes and antibody-free signal amplification to visualize miRNAs while preserving tissue morphology and spatial resolution. This approach could be adapted for simultaneous detection of multiple miRNAs. For complete details on the use and execution of this protocol, please refer to Harsh et al.¹.