This study explored the possible beneficial effects of singing on well-being during a singing lesson. Eight amateur (2m, 6f, age 28-53 yrs) and eight professional (4m, 4f, age 26-49 yrs) singers who had been attending singing lessons for at least six months were included. Continuous ECG was recorded and computerized spectral analysis was performed. Serum concentrations of TNF-alpha, prolactin, cortisol, and oxytocin were measured before and 30 min after the lesson. Five visual analogue scales (VAS, sad-joyful, anxious-calm, worried-elated, listless-energetic, and tense-relaxed) were scored before and after the lesson. In addition, a semi-structured interview was performed. Heart rate variability analyses showed significant changes over time in the two groups for total power, and low and high frequency power. Power increased during singing in professionals, whereas there were no changes in amateurs. This indicates an ability to retain more "heart-brain connection," i.e., more cardio-physiological fitness for singing in professional singers, compared to amateur singers. Serum concentration of TNF-alpha increased in professionals after the singing lesson, whereas the concentration in amateurs decreased. Serum concentrations of prolactin and cortisol increased after the lesson in the group of men and vice versa for women. Oxytocin concentrations increased significantly in both groups after the singing lesson. Amateurs reported increasing joy and elatedness (VAS), whereas professionals did not. However, both groups felt more energetic and relaxed after the singing lesson. The interviews showed that the professionals were clearly achievement-oriented, with focus on singing technique, vocal apparatus and body during the lesson. The amateurs used the singing lessons as a means of self-actualization and self-expression as a way to release emotional tensions. In summary, in this study, singing during a singing lesson seemed to promote more well-being and less arousal for amateurs compared to professional singers, who seemed to experience less well-being and more arousal.