In the case of tree species delineation with very high spatial resolution (VHR) images, is desirable that each segment corresponds to one individual tree crown (ITC). However, in order to have a segmentation algorithm that generates segments matching to ITCs, its parameters ought to be properly tuned. Aiming to avoid time-consuming trial-and-error procedures associated with this task, some initiatives for the automatic search of segmentation parameters have been developed, such as metaheuristic methods. The objective of this work was to test the automatic tuning of segmentation parameters of three segmentation algorithms for the delineation of ITCs belonging to a native endangered species in a subtropical forest area, comparing this method with the traditional trial-and-error approach. Two datasets (WorldView-2 and an orthoimage) and three segmentation algorithms (multiresolution, mean-shift and graph-based) were tested. For the automatic approach, a hybrid metaheuristic method was applied to accomplish the automatic search of parameters for the segmentation algorithms, while for the trial-and-error, a visual assessment was conducted for each set of parameters tested. Four supervised metrics were used to assess the quality of the segmentation results for the optimization approach and for the final set of parameters chosen in the trial-and-error approach. Results showed that none of the algorithms, datasets or approaches differ too much. The evaluation metrics values were lower, indicating that the reference ITCs polygons matched with the segmentation results. Despite the similar results, the automatic tuning of segmentation parameters proved to be a feasible alternative to reduce the subjectivity and the human effort in the choice of segmentation parameters as compared to the trial-and error approach.