Current codes for fire protection of buildings are mainly based on the movement of adults and neglect the movement characteristic of pre-school children. Having a profound comprehension of the difference between children and adults passing bottlenecks is of great help to improve the safety levels of preschool children. This paper presents an experimental study on the bottleneck flow of pre-school children in a room. The movement characteristics of children’s and adults’ bottleneck flow are investigated with two macroscopic properties: density and speed profiles as well as microscopic characteristic time: motion activation time, relaxation time, exit travel time and headway time. Arch-like density distributions are observed both for highly motivated children and adults, while the distance between the peak density region and the exit location is shorter for children and longer for adults. Children’s movement is less flexible manifested as longer motion activation time and longer relaxation time compared to that of adults. The findings from this study could enhance the understanding of crowd dynamics among the children population and provide supports for the scientific building design for children’s facilities.