Aims: To assess preschool children's attitudes towards their dentists. Methods: A questionnaire was developed to evaluate children's attitudes towards their dentists. Informed consent to distribute the questionnaire was obtained from the school principals and parents. The investigators filled the questionnaire out with the children. The questionnaire recorded age, gender, place of residence and contained 17 attitudinal items related to the dentist's gender, attire, age, personal protective equipment and dental clinic interior. The children expressed their attitudes to the above items by choosing from pictograms or photographs. A previous dental experience, if any, was recorded, along with the date of the first dental visit provided by the parents. In a pilot study, 44 children were asked to complete the questionnaire and to identify any questions which were unclear. Subsequently, the questionnaire was modified accordingly and retested for internal consistency (Cronbach's alpha 0.75). The child subjects for the main study were recruited in randomly selected nurseries in the capital city and rural regions. Then all children were included based on signed informed consent. Chi-square test at the significance level of 5% was used to test the relationships between the variables. Results: The questionnaire was completed by 439 children (247 girls, 192 boys; response rate 76.6%) within an age range of 3-7 years, attending public nurseries in the capital city Prague (312 children in 4 nurseries) and in other regions (127 children in 4 nurseries). In total, 376 (85.6%) of the study subjects had already been to the dentist. The girls preferred a dental office decorated with motifs for children more than boys (p=0.05). The children with a previous dental experience (p=0.05) and those who lived out of Prague (p=0.04) were more likely to prefer motifs for children in a dental office. The girls preferred female dentists at the age of their parents (p=0.05). The children currently undergoing dental treatment preferred female dentists, at the age of their parents (p=0.03), wearing colourful medical uniforms. Children from non-Prague regions preferred female dentist in coloured clothes without any protective equipment more than children from Prague (p=0.04). Several reasons for children's dental anxiety were identified. These were related to tooth drilling more in girls than in boys (p=0.03). The children preferred their parents staying in the dental office during their treatment (p=0.02). The children who lived out of Prague were more afraid of tooth drilling and extraction than those from Prague (p=0.03). Conclusions: The study described Czech preschool children's perceptions and preferences towards dental visits and dentists. Dental anxiety of children was related to the age of their first dental visit (2.5 years in non-Prague children, 1.6 years in Prague children), previous dental treatment and place of residence. Identification of children's attitudes towards their dentists could help initiate positive changes to make dental treatment more comfortable for preschool children. Acknowledgements: Supported by program PRVOUK-P 28/LF1/6. Keywords: preschool children, dental care, questionnaire, dentist, dental fear, dental anxiety