There is no exaggeration to assert that vaccination is one of the greatest scientific discoveries ever made. Generally, mothers’ misconception of vaccination was considered as one of the major barriers for better immunization coverage. Previous studies showed misconceptions on mothers’ knowledge towards childhood immunization. So, there is a necessity for the mothers to create unequivocal knowledge in order to develop positive attitude towards vaccination and thus improve their contribution to it for reducing the burden of dreadful infectious diseases. Objective: To develop and validate an educational booklet for treating mothers' misconceptions on vaccination. Design: A cross-sectional descriptive study design was adopted in this study for building and validating an educational booklet entitling "Treating mothers' misconceptions on vaccination." The construction of the booklet took place between June to December 2017 at Tanta district health offices through two stages: bibliographic survey; and manufacture of educational materials. Total sample size of 400 mothers of children under five years selected by using convenience sampling technique. Two-structured questionnaires were developed based on relevant literature and adopted throughout the study phases. The first one contained the following data: A) the first part included socio-demographic characteristics of the mothers as age, educational level, occupation, family size and residence. B) The second part included mothers' knowledge towards obligatory vaccination importance, contraindications, and side effects, in addition to all vaccines' mode, time and age of administration. C) The third part included mother’s practical knowledge she followed before, during and after vaccinating her child. Moreover, the second questionnaire was used for exploring the mothers and professional experts' opinions in relation to formulation of an educational booklet. Results: The study results indicated that, Out of 400 mothers included in the study, the mean age of the mother whose child undergoing vaccination is (22.00 ± 10.68) years. The predominant mothers have completed higher education, and occupational status showed that more than two-third (68.3%) were house wives. Regarding mothers' knowledge level, The proportion of mothers who lack the knowledge were as follows; vaccination contraindication (86.3%),Side effects (60.8%), BCG (51.5%), Hepatitis B (87%), MMR (92.5%), OPV (54.8%) and PENTA (94.5%). Conclusion: It was crucial to construct this booklet to ensure children' mothers' better understanding of the issues surrounding vaccination, treating their misconceptions and for improving their potentials of management of their children' vaccination care.