Simple Methods to Acutely Measure Multiple Timing Metrics among Sexual Repertoire of Male Drosophila

Male Drosophila courtship behavior is a key model for studying temporal decision-making. While courtship index (CI) is widely used to quantify mating activity, other timing metrics like courtship latency, copulation latency (CL), and mating duration (MD) remain understudied. Traditional methods for quantifying these behaviors are often labor-intensive and prone to human error. In this study, we present a protocol combining a modular chamber system and automated software (DrosoMating) to quantify 6 key timing metrics during male courtship. High-resolution video tracking enable precise analysis of courtship initiation, persistence, and mating dynamics under controlled conditions. Validation shows