Healthcare-associated infections (HAIs) are considered to be one of the biggest health problems as they continue to be an important cause of morbidity and mortality worldwide. They cannot be completely prevented, but their incidence can be significantly limited. Preventive action is the most important measure in this case. Due to the frequent interaction between healthcare professionals and patients, the crucial importance of hand hygiene is therefore emphasised. Adherence to good disinfection and hand washing practices remains around 40%, which can be improved by using a variety of nudge tools to promote desired hygienic behaviour. We conducted an open observation of employees and visitors with participation. The aim of this study was to determine the actual status of hand disinfection in a selected healthcare facility amongst doctors, registered nurses, medical technicians, cleaners, and visitors or parents of children; then, we selected and introduced three nudge tools of desired hygiene behaviour and analysed their effectiveness; finally, we provided suggestions for the use of nudge tools of desired hygiene behaviour with the aim of influencing doctors, registered nurses, medical technicians, cleaners, and visitors or parents of children so that they disinfect their hands properly. The actual state of hand disinfection was determined on the basis of observation without introducing any changes; then, we separately introduced three nudge tools, posters with an inscription and picture, the scent of citrus, and flashing lights. The obtained results were analysed with the help of the SpeedyAudit Lite application, and the effectiveness of each nudge tool and the adequacy of hand disinfection by categories of people were compared. In general, posters with a picture and an inscription contributed the most to more consistent disinfection of employees’ hands, while the scent of citrus and flashing lights contributed slightly less.