The discussion of the existence and measurement of layers in forests refers to one of the earliest and most controversial concepts of forest ecology, the stratification. Since there is no consensus on the most appropriate methodology to describe the vertical structure of forest communities, we chose for this paper to develop a methodology that was adequate to represent the stratification observed on site. The objective was to determine the species and functional traits characteristic of each vertical layer in the semideciduous seasonal forests (SSF). The study was conducted in ten fragments (10 ha) located in southeastern Brazil. Stratification was performed according to species using the median and the 3rd quartile (non-parametric statistical analysis) of tree heights were used for canopy stratification the (understory, midstory and canopy). This result shows the small range of the midstorey layer, highlighting the dichotomy between the canopy and understory. The variations found for the quartile and median values represent the history of successional stage of each fragment, allowing variations in the vertical occupation by species of certain layers. The analysis of SSF vertical structure allowed a visualization of the division of species and their respective functional traits performing different ecosystem functions in each layers.