Pulmonary vein (PV) stenosis following catheter ablation of atrial fibrillation (AF) is a new clinical syndrome. The optimal method of assessing this syndrome is not known. We evaluated radionuclide perfusion imaging, anatomic imaging, and direct measurements of PV-left atrial (LA) pressure gradients in patients suspected of having PV stenosis after catheter ablation for the treatment of AF. The study included 11 consecutive patients who were referred to a tertiary referral center for the evaluation of symptoms suggesting or imaging evidence of PV stenosis following catheter ablation for AF. All patients underwent anatomic imaging of their PVs with direct pulmonary venography or CT scanning as well as radionuclide perfusion imaging. PV stenosis (> 50% diameter) was diagnosed by venography in 6 of the 11 patients and in 16 of 44 PVs. All six patients with PV stenosis had perfusion defects in the affected pulmonary lobe. In contrast, all of the patients without anatomic evidence of PV stenosis had normal perfusion. There were 14 PVs with stenoses of > 80% of the luminal diameter, all of which had a corresponding perfusion abnormality ascertained by perfusion scanning. In all 14 PVs with a resting PV-LA gradient of > 5 mm Hg, there was a corresponding perfusion defect. PV stenosis results in decreased perfusion in the affected lobe when the resting PV-LA pressure gradient is at least 5 mm Hg or when there is 80% luminal stenosis. A perfusion scan may serve as an effective screening tool for PV stenosis and may be most useful in assessing the hemodynamic significance of an anatomic PV stenosis.