Objective: In the detection of seizure onset zones, arterial spin labeling (ASL) can overcome the limitations of positron emission tomography (PET) with F-18-fluorodeoxyglucose (F-18-FDG), which is invasive, expensive, and radioactive. PET/magnetic resonance (MR) systems have been introduced that allow simultaneous performance of ASL and PET, but comparisons of these techniques with stereoelectroencephalography (SEEG) and comparisons among the treatment outcomes of these techniques are still lacking. Here, we investigate the effectiveness of ASL compared with that of SEEG and their outcomes in localizing mesial temporal lobe epilepsy (MTLE) and assess the correlation between simultaneously acquired PET and ASL. Methods: Between October 2016 and August 2017, we retrospectively studied 12 patients diagnosed with pure unilateral MTLE. We extracted and quantitatively computed values for ASL and PET in the bilateral hippocampus. SEEG findings and outcome were considered the gold standard of lateralization. Finally, the bilateral asymmetry index (AI) was calculated to assess the correlation between PET and ASL. Results: Our results showed that hypoperfusion in the hippocampus detected using ASL matched the SEEG-defined epileptogenic zone in this series of patients. The mean normalized voxel value of ASL in the contralateral hippocampus was 0.97 +/- 0.19, while in the ipsilateral hippocampus, it was 0.84 +/- 0.14. Meanwhile, significantly decreased perfusion and metabolism were observed in these patients (Wilcoxon, p < 0.05), with a significant positive correlation between the AI values derived from PET and ASL (Pearson's correlation, r= 0.74, p < 0.05). Significance: In our SEEG- and outcome-defined patients with MTLE, ASL could provide significant information during presurgical evaluation, with the hypoperfusion detected with ASL reliably lateralizing MTLE. This non-invasive technique may be used as an alternative diagnostic tool for MTLE lateralization.