The aim of this study was to assess the correlation between technetium-99m methoxyisobutylisonitrile (MIBI) uptake by parathyroid adenomas, oxyphil cell content and volume of the lesions. Thirty-one patients with parathyroid adenomas were evaluated prospectively. Preoperative double-phase 99mTc-MIBI scintigraphy was performed in all patients and tracer uptake by parathyroid lesions was assessed semi-quantitatively employing region of interest ratios to normal adjacent neck areas. Surgical specimens underwent histological evaluation and oxyphil cell content was determined. The intensity of tracer uptake was compared with oxyphil cell content, volume of the lesions and serum levels of calcium and parathormone. 99mTc-MIBI tracer uptake was correlated with oxyphil cell content, volume of parathyroid lesions and the functional status of the parathyroid adenomas. Tracer accumulation in oxyphil cells might partially explain the preferential 99mTc-MIBI retention in parathyroid lesions.