Diffusion-weighted imaging (DWI) based thermometry has the potential to be a non-invasive method of temperature measurement for the deep inside of human brain. Nevertheless, the DWI at lateral ventricle in brain might be influenced by the pulsation flow of cerebrospinal fluid (CSF), which is motivated by heartbeat. The purpose of this study was to investigate the influence of pulsation flow on brain DWI thermometry for healthy subjects. Comparisons were performed between magnetic resonance spectroscopy and DWI based thermometry (ΔT) at three CSF speed selections. There was no significant difference on ΔT among the CSF speed and volume on healthy subjects.