The purpose of the study was to investigate the effects of shift work on the psychomotor and postural performance of night workers. The study included 20 polysomnography technicians working schedule of 12-h night shift by 36-h off. On the first day of protocol, the body mass and height were measured, and an actigraph was placed on the wrist of each participant. On the second day of protocol, sleepiness by Karolinska Sleepiness Scale, postural control by force platform (30 seconds) and psychomotor performance by Psychomotor Vigilance Task (10 minutes) were measured before and after 12-h night work. Results showed that after 12-h night work, sleepiness increased by 59% (p<0.001), postural control variables increased by 9% (p = 0.048), and 14% (p = 0.006). Mean reaction time, and the number of lapses of attention increased by 13% (p = 0.006) and 425% (p = 0.015), respectively, but the mean reciprocal reaction time decreased by 7%. In addition, there were correlations between sleepiness and postural control variables with opened eyes (r = 0.616, 95% confidence interval [CI] = 0.361–0.815; r = 0.538; 95% CI = 0.280–0.748) and closed eyes (r = 0.557; 95% CI = 0.304–0.764, r = 0497; 95% CI = 0.325–0.715) and a pronounced effect of sleepiness on postural sway (R2 = 0.393; 95% CI = 0.001–0.03). Therefore, 12-h night work system and sleepiness showed a negative impact in postural and psychomotor vigilance performance of night workers. As unexpected, the force platform was feasibility to detect sleepiness in this population, underscoring the possibility of using this method in the workplace to prevent occupational injuries and accidents.