INTRODUCTION: The instructional video "Be sweet to babies" was developed in Canada as a knowledge translation tool directed to parents on the use of analgesic strategies during painful procedures performed in newborns. AIM: To assess the effect of the Portuguese version of the "Be sweet to babies" video on maternal involvement in neonatal pain management during painful procedures. METHODS: A non-randomized, pragmatic clinical trial conducted in the rooming-in unit of the University of São Paulo affiliated hospital in July 2017. The study included 73 mothers, allocated in the control group (CG) (n=38) and in the experimental group (EG) (n=35). All participants received a pamphlet regarding neonatal pain management at the time of admission in the rooming-in. The EG was composed by mothers who voluntarily watched the video during daily educational sessions taught by nurses. Data were collected by means of interviews. The primary outcome was the prevalence of analgesic strategies during newborn screening test. The secondary outcomes were: feasibility, acceptability and usefulness of the instructional video; type of mothers' involvement during this procedure; analgesic strategies used (breastfeeding, skin-to-skin care and sweet tasting solution); pain intensity and newborn behavior; maternal feelings; satisfaction with pain control and information received. The study was conducted according to the 466/2012 Resolution, under approval of the Research Ethics Committees of the involved institutions, and after registration in the Brazilian Registry of Clinical Trials platform. RESULTS: Groups were homogeneous regarding maternal and neonatal characteristics, except for the mean number of children, which was higher in the CG. The "Be sweet to babies" video was considered as useful, easy to understand and easy to apply in real scenarios, with ideal length, and recommendable to other parents. Mothers of both groups were involved in newborns pain management. However, analgesic strategies were used in 40.0% of cases in the EG and 23.7% in the CG. There was a clinically significant difference of 16.3 percentage points between groups, although no statistical significance was found. The use of breastfeeding prevailed, followed by skin-to-skin care and non-nutritive sucking. In an adjusted analysis by other variables, the video increased the chance of analgesic strategies use by 2.1 times, while the nurse suggestion increased the chance of analgesic strategies use by 5.5 times. There was no difference between the experimental and control groups regarding pain intensity and newborns' behavior perceived by mothers. There was also no difference between groups with respect to maternal feelings during the procedure. Most of the mothers were satisfied with the nursing care in controlling the newborns' pain, however there was greater satisfaction with the amount of information received on neonatal pain management in the EG. Finally, almost all participants stated that they intend to use analgesic strategies in future painful procedures. CO...