Monitoring forest dynamics at an individual tree scale is essential for accurately assessing ecosystem responses to climate change, yet traditional methods relying on field-based forest inventories are labor-intensive and limited in spatial coverage. Advances in remote sensing using drone-acquired RGB imagery combined with deep learning models have promised precise individual tree crown (ITC) segmentation; however, existing methods are frequently validated against human-annotated images, lacking rigorous independent ground truth. In this study, we generate high-fidelity validation labels from co-located Terrestrial Laser Scanning (TLS) data for drone imagery of mixed unmanaged boreal and Mediterranean forests. We evaluate the performance of two widely used deep learning ITC segmentation models - DeepForest (RetinaNet) and Detectree2 (Mask R-CNN) - on these data, and compare to performance on further Mediterranean forest data labelled manually. When validated against TLS-derived ground truth from Mediterranean forests, model performance decreased significantly compared to assessment based on hand-labelled from an ecologically similar site (AP50: 0.094 vs. 0.670). Restricting evaluation to only canopy trees shrank this gap considerably (Canopy AP50: 0.365), although performance was still far lower than on similar hand-labelled data. Models also performed poorly on boreal forest data (AP50: 0.142), although again increasing when evaluated on canopy trees only (Canopy AP50: 0.308). Both models showed very poor localisation accuracy at stricter IoU thresholds, even when restricted to canopy trees (Max AP75: 0.051). Similar results have been observed in studies using aerial LiDAR data, suggesting fundamental limitations in aerial-based segmentation approaches in closed canopy forests.

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