The aim of the investigation was to study the significance of mat and shoe softness during prolonged work in an upright position based on some physiological, biomechanical and comfort measurements related to the lower extremities and the low back. Eight healthy female volunteers performed 2 h of simulated standing and 2 h of standing/walking work tasks in the laboratory using four combinations of soft shoes, clogs, soft mat and concrete. Thus, each subject performed a total of eight 2 h work tests. The following parameters were measured pre-experimentally and one or more times during 2 h: total foot volume, vascular volume and interstitial volume of the left foot, EMG from the lumbar paraspinals, movement of centre of gravity (only during standing), biomechanical heel impact (only during standing/walking), perceived discomfort in lumbar back, legs and feet, whole body oxygen uptake, arterial blood pressure and heart rate. Using soft shoes rather than clogs during standing/walking work implies approximately a halving of the foot oedema formation and the heel impact. The effects due to the introduction of the soft mat are negligible. The local circulatory responses in the feet and the EMG-signs of paravertebral muscle fatigue are larger during standing compared to standing/walking work. The two investigated work types in this study differ regarding exposures as well as responses. Thus, it is recommended to shift between these postures and seated work during the working hours to improve job exposures.