Purpose: We conducted a comparative study to evaluate the diagnostic values of Tl-201, Tc-99m MIBI and I-131 scans in the follow-up assessment after ablative I-131 therapy in differentiated thyroid cancer. Materials and Methods: The study population consisted of 20 patients who underwent surgical removal of thyroid cancer and ablative radioactive iodine therapy, and followed by one or more times of I-131 retreatments (33 cases). In all patients, Tl-201, Tc-99m MIBI, diagnostic and therapeutic I-131 scans were performed and the results were analyzed retrospectively. Also serum thyroglobulin levels were measured in all patients. The final diagnosis of recurrent or metastatic thyroid cancer was determined by clinical, biochemical, radiologic and/or biopsy findings. Results: Positive rates (PR) of Tc-99m MIBI, Tl-201, diagnostic and therapeutic I-131 scans in detecting malignant thyroid tissue lesions were 70% (19/27), 54% (15/28), 35% (17/48) and 63% (30/48), respectively. The PR in the group of 20 cases (28 lesions) who underwent concomitant Tl-201 and I-131 scans were in the order of therapeutic 131 scan 71%, Tl-201 scan 54% and diagnostic I-131 scan 36%. There was no statistically significant difference between Tl-201 and diagnostic I-131 scans (p>0.05). In the group of 20 cases (27 lesions) who underwent concomitant Tc-99m MIBI and I-131 scans, the PR were in the order of Tc-99m MIBI scan 70%, I-131 therapeutic scan 52% and I-131 diagnostic scan 33%. The PR of Tc-99m MIBI was significantly higher than that of diagnostic I-131 scan (p<0.05). Conclusion: Tc-99m MIBI scan is superior to diagnostic I-131 scan in detecting recurrent or metastatic thyroid cancer following ablation therapy in patients with differentiated thyroid cancer. Tl-201 scan did not showed significantly higher positive rate than diagnostic I-131 scan. Instead of diagnostic I-131 scan before the I-131 retreatment, Tc-99m MIBI scan without discontinuing thyroid hormone replacement would be a prudent and effective approach in the management of these patients.