We studied the diagnostic value of 16 slices of single photon emission computed tomography (SPECT)/computed tomography (CT) in the anatomical localization, image interpretation and extra-sentinel lymph nodes (SLNs) detection compared to dynamic and static planar radioisotopic lymphoscintigraphy (PLS) in patients with melanoma. Eighty-two patients with melanoma underwent dynamic PLS, static PLS and SPECT/CT. Data were obtained using a dual head SPECT/CT 16 slices γ-camera. We evaluated the number and localization of SLNs detected with each imaging method. SPECT/CT demonstrated 48 additional SLNs in comparison with PLS in 29 patients. In five truncal and seven head-neck lesions, dynamic and static PLS failed to detect the SLNs found on SPECT/CT (false negative). In one case of truncal and one case of lower limb melanoma, the foci of increased activity interpreted on PLS as possible SLNs were confirmed to be non-nodal sites of uptake on SPECT/CT (false positive). PLS underestimated the number of SLNs detected, whereas SPECT/CT revealed higher agreement compared to the respective number from histological reports. SPECT/CT showed a better prediction of the number of SLNs and higher diagnostic parameters in comparison to planar imaging. SPECT/CT is an important complementary diagnostic modality to PLS, that improves detection, preoperative evaluation, anatomical landmarks of SLNs and surgical management of patients with melanoma.