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The Trier Social Stress Test (TSST) is a widely used, reliable, and ecologically valid method for inducing acute stress under controlled conditions. Traditionally, the TSST is administered with staff physically present with participants, which limits the participant populations that can be exposed to the TSST. We describe an adaptation of the TSST to remote, online delivery over video-conferencing, which we call the internet-delivered Trier Social Stress Test (iTSST). This adaption has participants use wearable, self-administered ECG monitors received and returned via mail. Fifty participants were recruited to take part in a pilot study evaluating stress-reduction interventions and completed the iTSST at two occasions separated by approximately 12 weeks. Perceived stress and heart rate variability (HRV) were measured during both administrations of the iTSST. Forty-one participants completed both assessments and were included in the set of analyses. Both administrations were characterized by an increase in self-reported stress and reduction in self-reported relaxation from the resting phase to the speech task, which returned to baseline during recovery. In terms of HRV, we observed a significant parasympathetic response to the iTSST in 90% of participants, evidenced by a decrease in RMSSD and increase in heart rate from resting to the speech task, which recovered during the recovery phase. In terms of repeatability, there was little evidence of habituation and the iTSST elicited a stress response during both the initial administration and the 12-week follow-up. While the utility is limited by the lack of a measure of sympathetic and HPA-axis activity, the iTSST represents a promising research tool when physically interacting with participants is not feasible. • The Trier Social Stress Test was adapted to an online environment (iTSST). • Parasympathetic withdrawal in response to the iTSST occurred among 90% of participants. • iTSST reliably increased self-reported stress and reduced relaxation. • Little evidence of habituation to the iTSST was observed over 12-weeks time. • iTSST may be a cost effective and ecologically valid ambulatory stress test. [ABSTRACT FROM AUTHOR]