The amenability to genetics ofDrosophila melanogasterhas made this organism one of the best-suited models for studying the neurobiology of pheromone-guided behavior. Single-male assays use the minigene encoding the thermosensitive channeldTrpA1to activate neurons expressingfruitless(fru), a major courtship regulator gene, and thereby induce most of the elementary courtship acts in a solitary male exposed to temperature increase. Tethered male assays allow Ca2+ -imaging of neuronal activities of a male fly displaying courtship behavior on a treadmill when stimulated with a female or pheromones. Here we describe technical details of these assays.