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Combining physiological measures with observational data (e.g., video or self-reports) to further capture and understand the temporal and cyclical process of social regulation has become a trend in the field. Synchronized physiological arousal is a particularly meaningful situation in collaboration. However, little attention has been given to synchronized physiological arousal episodes and their relationship with the social regulatory process. In addition, only a few research utilized heart rate (HR) as a physiological measure in the current collaboration literature. More research is necessary to reveal the potential of HR to expand the diversity of physiological indicators in the field. Therefore, the current study aimed to explore what synchronized physiological arousal can further reveal about the social regulatory process. To achieve this goal, this study designed a collaborative argumentation (CA) activity for undergraduates (mean age 20.3). It developed an arousal-regulation analysis platform, which could automatically detect synchronized physiological arousal in HR and align them with coding challenges and social regulation based on the timeline. In total, 14 four-member groups were recruited. After analyzing both videos and HR data, several findings were obtained. First, only one-third of episodes were synchronized physiological arousal episodes, and the situations where four members were all in arousal states were rare during CA. Second, synchronized physiological arousal was more sensitive to socio-emotional aspects of collaboration as the shared physiological arousal more frequently co-occurred with socio-emotional challenges and socio-emotional regulation, while it happened the least under motivational challenges. Third, synchronized physiological arousal has also been found to be associated with the challenges being regulated. Finally, pedagogical implications were suggested., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Li, Hu, Li and Mao.)