Background Flow-related signal void artifacts can be visualized on the T2-weighted (T2W) three-dimensional sampling perfection with application-optimized contrast (3D-SPACE) sequence. Flow void artifacts in the cerebral aqueduct and the fourth ventricle can provide information about cerebrospinal fluid (CSF) flow dynamics. Purpose In this study, we aimed to test the performance of the T2W 3D-SPACE sequence in assessing the CSF flow in the aqueduct and/or fourth ventricle. Material and Methods A total of 137 patients (age range = 3–89 years) who underwent CSF flow study were included. The amount of signal loss on T2W 3D-SPACE due to flow in the aqueduct and fourth ventricle was assessed and graded using a 4-point scale of 0 (absence of flow void) to 3 (signal void filling the aqueduct and entire fourth ventricle). A correlation was then sought between the quantitative values obtained by phase-contrast magnetic resonance imaging (PC-MRI) and the amount of signal void in the 3D-SPACE sequence. Results At the aqueduct level, there was a statistically significant difference in the forward flow velocity and the flow volume among different grades (all P < 0.001). In the grade 3 group, CSF peak systolic flow velocity and mean flow volume were found to be significantly higher than in the other grades ( P < 0.001). The mean aqueduct area in the grade 0 group was found to be significantly different from that in the other classes ( P < 0.001). Conclusion The amount of signal loss in the fourth ventricle observed on T2W 3D-SPACE is correlated with the peak systolic velocity and flow volume measured quantitatively in PC-MRI.