BACKGROUND: We have acquired dual-echo spin-echo (DE SE) MRI data of the rhesus monkey brain since 1994 as part of an ongoing study of normal aging. To analyze these legacy data for regional volume changes, we have created a reference label atlas for the Template Driven Segmentation (TDS) algorithm. METHODS: The atlas was manually created from DE SE legacy MRI data of one behaviorally normal, young, male rhesus monkey and consisted of 14 regions of interest (ROI’s). We analyzed the reproducibility and validity of the TDS algorithm using the atlas relative to manual segmentation. RESULTS: ROI volumes were comparable between the two segmentation methodologies, except TDS overestimated the volume of basal ganglia regions. Both methodologies were highly reproducible, but TDS had lower sensitivity and comparable specificity. CONCLUSIONS: TDS segmentation calculates accurate volumes for most ROI’s. Sensitivity will be improved in future studies through the acquisition of higher quality data.