Objective: The study aimed to evaluate the interobserver agreement among glaucoma subspecialists in diagnosing glaucoma and to explore the causes of diagnostic discrepancies. Methods: Three experienced glaucoma subspecialists independently assessed frequency domain optical coherence tomography, fundus color photographs, and static perimetry results from 464 eyes of 275 participants, adhering to unified glaucoma diagnostic criteria. All data were collected from the Wenzhou Glaucoma Progression Study between August 2014 and June 2021. Results: The overall interobserver agreement among the three experts was poor, with a Fleiss’ kappa value of 0.149. The kappa values interobserver agreement between pairs of experts ranged from 0.133 to 0.282. In 50 cases, or approximately 10.8%, the three experts reached completely different diagnoses. Agreement was more likely in cases involving larger average cup-to-disc ratios, greater vertical cup-to-disc ratios, more severe visual field defects, and thicker retinal nerve fiber layer measurements, particularly in the temporal and inferior quadrants. High myopia also negatively impacted interobserver agreement. Conclusions: Despite using unified diagnostic criteria for glaucoma, significant differences in interobserver consistency persist among glaucoma subspecialists. To improve interobserver agreement, it is recommended to provide additional training on standardized diagnostic criteria. Furthermore, for cases with inconsistent diagnoses, long-term follow-up is essential to confirm the diagnosis of glaucoma.