Automated Behavioral Tracking and Analysis of Xenopus Tadpoles.

Automated behavioral tracking provides a quantitative and reproducible way to measure locomotor activity in Xenopus tadpoles. We describe a method for simultaneously recording 24-free swimming tadpoles in 24-well plates under back-light illumination using a Raspberry Pi-based computer and camera. Locomotion is analyzed using TopScan tracking software, which identifies the center mass for each tadpole within its arena. The protocol quantifies locomotor parameters such as swim distance, velocity, and activity patterns. We outline steps required to assemble and calibrate the imaging hardware, configure the recording script, and process video data for downstream behavioral analysis. Together, this workflow provides a reliable foundation for studying locomotor behavior and detecting alterations arising from genetic or pharmacological manipulations.