This is a postprint version of the following published document:Mateos-Pérez, J. M.; Soto-Montenegro, M. L.; Peña-Zalbidea, S.; Desco, M.; Vaquero, J. J. (2016). "Functional segmentation of dynamic PET studies: Open source implementation and validation of a leader-follower-based algorithm". Computers in Biology and Medicine, v. 69, February, pp. 181-188. DOI: 10.1016/j.compbiomed.2015.12 We present a novel segmentation algorithm for dynamic PET studies that groups pixels according to the similarity of their time activity curves. Methods: Sixteen mice bearing a human tumor cell line xenograft (CH 157MN) were imaged with three different 68 Ga DOTA peptides (DOTANOC, DOTATATE, DOTATOC) using a small animal PET CT scanner. Regional activities (input function and tumor) were obtained after manual delineation of regions of interest over the image. The algorithm was implemented under the jClustering framework and used to extract the same regional activities as in the manual approach. The volume of distribution in the tumor was computed using the Logan linear method. A Kruskal Wallis test was used to investigate significant differences between the manually and automatically obtained volumes of distribution. Results: The algorithm successfully segmented all the studies. No significant differences were found for the same tracer across different segmentation methods. Manual delineation revealed significant differ ences between DOTANOC and the other two tracers (DOTANOC DOTATATE, p¼0.020; DOTANOC DOTATOC, p¼0.033). Similar differences were found using the leader follower algorithm.Conclusion: An open implementation of a novel segmentation method for dynamic PET studies is pre sented and validated in rodent studies. It successfully replicated the manual results obtained in small animal studies, thus making it a reliable substitute for this task and, potentially, for other dynamic segmentation procedures.