Occupational stress is universally experienced and is emerging as a major risk factor for physical and mental illness and a key factor in poor work performance and low job satisfaction. However, the technology does not currently exist to unobtrusively measure occupational stress in real-time. Here, we describe the design and clinical validation of an automated high-definition thermal imaging system that can be used to quantify human stress, remotely and instantaneously. Healthy human subjects underwent a computer-based version of the Stroop-color conflict test, which is a validated stress provocation test, in an experimental office facility. In separate experiments, the same subjects completed a mental arithmetic challenge. The thermal signal associated with stress provocation is near-instantaneous corrugator warming. The stress response was detected in all subjects for all stress-events compared to the respective baselines. Furthermore, there was remarkable inter-individual preservation of the corrugator signal with stress R(2) = 0.96, P< 0.001). High-definition thermal imaging can be used for real-time detection of stress provocation. This technology may prove to be of help in ameliorating office-place stress.