PyRodentTracks: flexible computer vision and RFID based system for multiple rodent tracking and behavioral assessment.

PyRodentTracks (PRT) is a scalable and customizable computer vision and RFID-41 based system for multiple rodent tracking and behavior assessment that can be set up within 42 minutes in any user-defined arena at minimal cost. PRT is composed of the online Raspberry 43 Pi-based video and RFID acquisition and the subsequent offline analysis tools. The system is 44 capable of tracking up to 6 mice in experiments ranging from minutes to days. PRT 45 maintained a minimum of 88% detections tracked with an overall accuracy >85% when 46 compared to manual validation of videos containing 1-4 mice in a modified home-cage. As 47 expected, chronic recording in home-cage revealed diurnal activity patterns. Moreover, it was 48 observed that novel non-cagemate mice pairs exhibit more similarity in travel trajectory 49 patterns over a 10-minute period in the openfield than cagemates. Therefore, shared features 50 within travel trajectories between animals may be a measure of sociability that has not been 51 previously reported. Moreover, PRT can interface with open-source packages such as 52 Deeplabcut and Traja for pose estimation and travel trajectory analysis, respectively. In 53 combination with Traja, PRT resolved motor deficits exhibited in stroke animals. Overall, we 54 present an affordable, open-sourced, and customizable/scalable rodent-specific behavior 55 recording and analysis system. ### Competing Interest Statement The authors have declared no competing interest.