A behavioral assay to test sensory-cue-guided oriented flight in monarch butterflies under controlled conditions

Many animals use sensory cues to guide movement. Testing animals under conditions in which cues can be isolated and manipulated is key for understanding the function of cues. Here, we present a protocol to assess the flight of migratory monarch butterflies (Danaus plexippus). We describe procedures to optimize and conduct trials, especially under indoor conditions. This protocol facilitates testing monarchs in various experimental conditions including during their subjective night when they are not normally flying. For complete details on the use and execution of this protocol, please refer to Parlin et al. (2022).¹.