Studies report that 11 C-flumazenil (FMZ) PET more specifically localizes the epileptogenic zone in patients with medically refractory focal epilepsy than 18 F-FDG PET. However, practical aspects of 11 C use limit clinical application. We report a phase I/IIa study assessing the clinical use of 18 F-FMZ PET for the localization of the epileptogenic zone in patients with drug-resistant temporal lobe epilepsy (TLE). Receptor binding was quantified using kinetic modeling that did not require arterial sampling. Methods: Dynamic 18 F-FMZ PET and static interictal 18 F-FDG PET scans were compared in healthy controls (n 5 17 for 18 F-FMZ and n 5 20 for 18 F-FDG) and TLE patients with mesial temporal sclerosis on MR imaging (MTS, n 5 12) and with normal MR imaging (NL TLE, n 5 19). Masked visual assessment of images was undertaken. Parametric images of 18 F-FMZ binding potential (BP ND ) were generated using the simplified reference tissue model. Region-of-interest analysis on coregistered MR images and statistical parametric mapping were used to quantify 18 F-FMZ BP ND and 18 F-FDG uptake in the temporal lobe. Results: The visual assessment of static standardized uptake value images showed 18 F-FMZ PET to have high specificity (16/17 [94%]) and moderate sensitivity (21/31 [68%]) for the localization of the epileptogenic zone, with a more restricted abnormality than 18 F-FDG PET. However, the 18 F-FMZ standardized uptake value images were falsely localizing in 3 of 31 patients (10%). Region-of-interest analysis demonstrated reductions in ipsilateral hippocampal 18 F-FMZ BP ND in patients with either MTS or NL TLE, compared with controls subjects. Ipsilateral hippocampal 18 F-FMZ BP ND was independent of both hippocampal volume and 18 F-FDG uptake, whereas ipsilateral hippocampal volume was correlated with 18 F-FDG uptake (r 2 5 0.69, P , 0.0001). Statistical parametric mapping analysis demonstrated decreased uptake in 14 of 31 (45%) cases with 18 F-FMZ PET and 18 of 29 (62%) with 18 F-FDG PET. Cluster size was significantly smaller on 18 F-FMZ than 18 F-FDG images (37 vs. 160 voxels, P , 0.01). Conclusion: 18 F-FMZ PET has potential as a clinical tool for the localization of the epileptogenic zone in the presurgical evaluation of drug-resistant TLE, providing information complementary to 18 F-FDG PET, with a more restricted region of abnormality.