OBJECTIVE To evaluate pulsatile ocular blood flow (POBF) analysis in normal subjects and glaucoma patients by comparison of POBF measurements with functional (as determined by visual field [VF]) and structural (as determined by optical coherence tomography [OCT]) measures. METHODS Prospective, cross-sectional study. METHODS Forty-one eyes of 24 consecutive glaucoma patients and 20 eyes of 10 healthy subjects were studied; POBF analysis was performed on all subjects at the same visit as VF testing and OCT retinal nerve fiber layer (NFL) thickness measurement. The mean results of normal and glaucomatous eyes were compared for each method. Correlation between measurements obtained with each modality and the discriminating power using receiver operator characteristic curves was tested. RESULTS The mean POBF (standard deviation [SD]) in the normal group was 1,010.4 (292.8) microl/min and 989.3 (305.5) microl/min in the glaucoma group (P =.90). Significant differences between groups were found for VF mean deviation and pattern standard deviation (P =.02, P =.004, respectively) and OCT mean NFL thickness (P <.0001). No correlation was found between POBF parameters and intraocular pressure, VF, or OCT variables except for intraocular pressure in glaucoma group (r = -.43, P =.003). The area under the receiver operator characteristic curves was higher for VF indexes and OCT mean NFL thickness than POBF parameters for distinguishing between normal and glaucomatous eyes. CONCLUSIONS The wide range of normal values and the low discriminating power of POBF between normal and glaucomatous eyes limits the clinical use of the device for glaucoma patients.