Abstract Background Examining information sources, communication channels, and preferred content formats is crucial for adapting and improving future communication strategies in vaccination campaigns. This study aims to evaluate the information and communication preferences regarding the booster dose of Covid-19 among employees of the Central Tuscany Local Health Authority (CT-LHA). Methods From July to October 2022, a questionnaire was administered to 7,000 out of approximately 15,000 CT-LHA employees. The questionnaire analyzed communication, organizational, and access aspects regarding preferences for the Covid-19 booster dose. The sample was stratified into healthcare workers (HCWs) and non-healthcare workers (NHCWs). Chi-squared tests were performed using STATA, with the significance level set at 95%. Results The returned questionnaires were 1,885 (26.9%), with the majority being HCWs (64.4%). HCWs rated the quality of communication significantly lower compared to NHCWs, who rated it very high (p < 0.05). HCWs also rated the ease of booking the Covid-19 booster dose and access modalities to vaccination significantly lower than NHCWs (p < 0.05). NHCWs expressed a preference for receiving scientific information from Ministry of Health tutorials, while HCWs preferred information from their general practitioner (p < 0.05). HCWs preferred to receive organizational information from healthcare management, while NHCWs preferred information from vaccination physicians (p < 0.05). Additionally, HCWs preferred company operating procedures for disseminating information, whereas NHCWs preferred corporate communications circulars (p < 0.05). Conclusions HCWs and NHCWs showed differences in perceived quality of communication, ease of access to vaccination services, and preferred sources of information. HCWs rated the quality of communication and access to vaccination services lower than NHCWs. This results suggest the need for tailored communication strategies to effectively reach both groups. Key messages • Health organizations need to develop targeted communication approaches that address the specific needs and preferences of their employees. • Flexibility and adaptability are crucial to adapt communication methods and channels to strengthen vaccination initiatives and promote public health outcomes.