Three different doses (1.0, 1.5, and 2.0 ml) of iohexol (300 mgI/ml) were injected percutaneously into the popliteal lymph node of eight adult cats under ultrasound guidance. Serial transverse CT images of five regions of interest (L3, T13, T8, T4, and T1 level) were performed at 2-min intervals, and the attenuation in Hounsfield Units (HU) of the lymphatic vessels was measured for determination of the optimal dose of iohexol and CT scan parameters. The optimal dose was 1.5 ml and helical CT acquisition is recommended to be performed as soon as possible after iohexol injection. In helical scans, the thoracic duct was characterized by variable branch numbers that formed a single trunk and entered the venous system at variable levels. CT lymphography using this protocol was performed in a cat with chylothorax. The thoracic duct was tortuous and focally dilated, and leakage of contrast medium was observed. Percutaneous CT lymphography using ultrasound-guided administration of iohexol into the popliteal lymph node appears reliable for delineation of the thoracic duct in cats.