AbstractA novel 3D nnU-Net-based of algorithm was developed for fully-automated multi-organ segmentation in abdominal CT, applicable to both non-contrast and post-contrast images. The algorithm was trained using dual-energy CT (DECT)-obtained portal venous phase (PVP) and spatiotemporally-matched virtual non-contrast images, and tested using a single-energy (SE) CT dataset comprising PVP and true non-contrast (TNC) images. The algorithm showed robust accuracy in segmenting the liver, spleen, right kidney (RK), and left kidney (LK), with mean dice similarity coefficients (DSCs) exceeding 0.94 for each organ, regardless of contrast enhancement. However, pancreas segmentation demonstrated slightly lower performance with mean DSCs of around 0.8. In organ volume estimation, the algorithm demonstrated excellent agreement with ground-truth measurements for the liver, spleen, RK, and LK (intraclass correlation coefficients [ICCs] > 0.95); while the pancreas showed good agreements (ICC = 0.792 in SE-PVP, 0.840 in TNC). Accurate volume estimation within a 10% deviation from ground-truth was achieved in over 90% of cases involving the liver, spleen, RK, and LK. These findings indicate the efficacy of our 3D nnU-Net-based algorithm, developed using DECT images, which provides precise segmentation of the liver, spleen, and RK and LK in both non-contrast and post-contrast CT images, enabling reliable organ volumetry, albeit with relatively reduced performance for the pancreas.