Purpose: To investigate the acute effects of brinzolamide, betaxolol, and latanoprost (drugs commonly used in the medical management of glaucoma) on choroidal thickness using enhanced depth imaging optical coherence tomography (EDI-OCT). Methods: Ninety healthy volunteers were evaluated in this prospective study. Participants were randomly divided into 3 groups. Brinzolamide, betaxolol, and latanoprost were administered into the left eyes of the first group (n = 30), second group (n = 30), and third group (n = 30), respectively, and artificial tear (Sodium hyaluronate) was instilled into the right eyes of all participants. Subfoveal choroidal thickness (SFCT) was measured using EDI-OCT before and 45 minutes after administration of the antiglaucomatous drops. Results: SFCT revealed a significant increase in the left eye (administered antiglaucomatous drop) in the brinzolamide (p = 0.001) and betaxolol groups (p = 0.049) and a significant increase also in the right eye (administered artificial drop) in the brinzolamide (p = 0.001) and betaxolol groups (p = 0.001). However, SFCT did not reveal a significant increase in the left eye (p = 0.213) or in the right eye (p = 0.062) in the latanoprost group. Conclusion: Brinzolamide and betaxolol caused an increase in SFCT, while latanoprost had no significant effect on SFCT.