Stress among university students is a growing concern across the Netherlands, Europe, and the world, yet the influence of physical learning environments on stress remains underexplored. This thesis investigates how two contrasting university study environments influence students’ stress responses, both physiologically and psychologically. The study compares two real-life university rooms at Delft University of Technology: a traditional, warm-toned space and a newer, cool-toned, modern environment. Stress was measured using salivary cortisol, custom-designed survey items, and thematic analysis of open-ended survey responses. Results show that students in the newer room reported improved concentration and reduced self-perceived physiological stress. In contrast, cortisol levels slightly increased in this space, though not significantly. This rise may indicate focused arousal or “eustress” rather than distress, suggesting that environments supporting concentration might trigger physiological activation even when perceived stress is low. Qualitative responses supported this interpretation, with students describing task-dependent preferences and subconscious responses to room design. These findings suggest that the physical learning environment can influence how students experience stress, while also highlighting a mismatch between psychological and physiological indicators. The divergence reinforces the importance of using multi-method approaches in environmental stress research, implementing multiple stress indicators both physiological and psychological. The study concludes that a diverse range of adaptable study environments is needed to accommodate students’ varying emotional and cognitive needs. Physical design should be considered a meaningful factor in promoting student wellbeing and academic performance.