AIM: To investigate the influence of non-oil 95 (N95)/filtering facepiece 2 (FFP2) facemask wear on retinal thickness, choroidal thickness (CT), retinal nerve fiber layer thickness (RNFLT), and ganglion cell layer thickness (GCLT) in healthy subjects. METHODS: In this prospective study, 53 healthy participants who used FFP2/N95 facemask were enrolled. Participants underwent optical coherence tomography imaging before and at 1 and 4h following FFP2/N95 facemask wear. The last imaging session was performed 1h after FFP2/N95 removal. Retinal thickness, CT, RNFLT, and GCLT were assessed at each session. Vital parameters were also assessed. RESULTS: The pulse rate of the subjects significantly decreased at 1 and 4h compared to baseline values (P<0.05). No significant changes in retinal thickness, RNFLT, and GCLT were observed in the study. CT profile showed a significant increase at all measured locations except 1-mm temporal, 1-mm inferior and 2-mm inferior points following FFP2/N95 wear which turned to baseline values after FFP2/N95 removal. Pulse rate and CT changes at 4h were significantly correlated (P<0.05). CONCLUSION: Parasympathetic activation during FFP2/N95 facemask wear might have a role on elevated CT measurements in healthy individuals by virtue of increased choroidal blood flow.