Abstract Background Hand hygiene is universally recognized as a cornerstone measure for the prevention of healthcare-associated infections. Although the WHO “My five Moments for hand hygiene” poster has been used for more than a decade to delineate hand hygiene indications and promote action, adherence levels among healthcare workers are still notoriously low and disquieting. To compensate for the lack of effective hand hygiene communication, we aimed to evaluate emojis as possible surrogates for the non-verbal aspects of hand hygiene behaviour. Methods Following a thorough review of the Unicode version 12.0, the most applicable emojis to the terms used in the WHO 5 Moments poster were extracted. We developed a self-administered questionnaire to assess the view of infection prevention and control (IPC) practitioners regarding the use of emojis to show the WHO 5 Moments. Completed questionnaires were collected and analysed to determine the suitability of the existing emojis to illustrate a unified emoji poster. Data were analysed using R (version 3.6.3). Results A total of 95 IPC practitioners completed the questionnaire from May to October 2019 from different countries. Of these, 69 (74%) were female, and the mean age of the participants was 44.6 ± 10.87 years. We found appropriate emojis for six of the words used in the poster, including "Image missing" for touching (72%), "Image missing" for patient (63%), "Image missing" for clean (53%), "Image missing" for procedure (56%), "Image missing" for body fluid (58%), and "Image missing" for exposure risk (71%). The existing emojis proposed for the words “hygiene”, “aseptic”, and “surrounding” seemed to be less satisfactory. Conclusions In summary, the findings of this study indicate that the existing emojis may not be able to substitute the words used in the WHO 5 Moments poster. Emojis might be helpful to address hand hygiene indications in healthcare that may eventually play a role in promoting this measure. However, emojis should be further studied to choose the most appropriate ones and avoid ambiguity and misinterpretation. More emojis to convey health related messages are needed. We recommend further research in this area to evaluate the effect of using emojis in healthcare-related behaviours.