Objectives <p>Compassion is a valuable, trainable skill which can bring significant benefits to oneself and others. One method for developing compassion towards others is Tonglen, a Tibetan Buddhist meditation which involves taking in suffering from others and sending them well-being. The aim of this study was to investigate the psychophysiological outcomes of Tonglen meditation in healthcare workers, a population who have frequent contact with the suffering of others.</p> Method <p>Sixty participants were randomly assigned to listen to a 15-min audio of either guided Tonglen meditation, or a story in the control condition, and completed assessments before and after.</p> Results <p>The results showed that, compared to the control condition, Tonglen significantly increased heart rate variability, compassion state, and affective responses to suffering.</p> Conclusions <p>This is the first study to show the acute effectiveness of Tonglen in healthcare workers. Results suggest a potential general applicability of this active compassion meditation to populations experiencing empathic distress and burnout.</p> Preregistration <p>This study was preregistered in Open Science Framework (<a href="https://osf.io/x7yh4">https://osf.io/x7yh4</a>).</p>