This study aimed to evaluate toothbrushing practices and toothpaste dosing among preschool children aged 0-6 years in Switzerland recruited from the Department of General Pediatric and Adolescent Dentistry, University Center for Dental Medicine Basel UZB, and in daycare centers or in private practices located in the canton of Berne. Three hundred parents of children were surveyed about socioeconomic characteristics, their children's toothbrushing behavior, the use of age-appropriate toothpaste, and whether they were instructed by an oral healthcare professional about appropriate toothbrushing practices and the amount of toothpaste to use. Additionally, the parents were asked to apply toothpaste to two different toothbrushes, one with a narrow and one with a wide brush head, as they usually would at home. The amount of toothpaste was weighed using a portable scale. Over 50% of the parents were not instructed by an oral health care professional on toothbrushing practices and toothpaste dosing. However, nearly all participants used age-appropriate toothpaste. Approximately 50% of the parents of 0- to 2-year-old children applied more than the recommended 0.25 g of fluoridated toothpaste (both brushes, mean ± SD: 0.25 g ± 0.14 g), while two-thirds of the parents of 2- to 3-year-olds (both brushes, mean ± SD: 0.36 g ± 0.23 g) and nearly 90% of the parents of 3- to 6-year-olds applied more than 0.25 g (both brushes, mean ± SD: 0.43 g ± 0.20 g). Overall, parents dispensed more toothpaste on the toothbrush with the wide brush head. The present study suggested that assuming toothpaste as the sole source of fluoride, the amounts generally used are not of concern. However, considering the substantial variation in the amounts of toothpaste applied, oral healthcare professionals should routinely provide instructions on toothbrushing practices and toothpaste dosing during children's dental examinations.