Several authors have studied the influence of light on both human physiology and emotions. Blue light has been proved to reduce sleepiness by suppression of melatonin secretion and it is also present in many emotion-related studies. Most of these have a common lack of objective methodology since results and conclusions are based on subjective perception of emotions. The aim of this work was the objective assessment of the effect of blue lighting in post-stress relaxation, in comparison with white lighting, by means of bio-signals and standardized procedures. We conducted a study in which twelve healthy volunteers were stressed and then performed a relaxation session within a chromotherapy room with blue (test group) or white (control group) lighting. We conclude that the blue lighting accelerates the relaxation process after stress in comparison with conventional white lighting. The relaxation time decreased by approximately three-fold (1.1 vs. 3.5 minutes). We also observed a convergence time (3.5–5 minutes) after which the advantage of blue lighting disappeared. This supports the relationship between color of light and stress, and the observations reported in previous works. These findings could be useful in clinical and educational environments, as well as in daily-life context and emerging technologies such as neuromarketing. However, our study must be extended to draw reliable conclusions and solid scientific evidence.