Purpose: The purpose of this study was to evaluate the effect of pharmacologic pupillary dilatation on anterior chamber depth (ACD) and anterior chamber angle (ACA) in eyes with exfoliation syndrome (XFS). Methods: Thirty-six eyes of 36 patients with XFS were evaluated with slit-lamp examination, Goldmann applanation tonometry and ultrasound biomicroscopy (UBM) under standard light conditions. Primary outcome parameters were defined as the change in ACD and ACA measured before and 40 min after instillation of a single drop of either 1% cyclopentolate (Group I; n = 12), 2.5% phenylephrine (Group II; n = 12) or 1% tropicamide (Group III; n = 12). Change in intraocular pressure (IOP) during the same time interval was included as a secondary outcome measure. Results: The average predilatation ACD, ACA and IOP values in the study subjects were 2.54 ± 0.40 mm, 27.9 ± 6.3° and 14.9 ± 3.1 mmHg, respectively. There were no significant differences in the mean age (p = 0.461), the female/male ratio (p = 0.232), baseline ACD (p = 0.841), ACA (p = 0.761) or IOP (p = 0.070) within the three groups. Differences in dilation induced changes in ACD (p = 0.108), ACA (p = 0.636) and IOP (p = 0.160) between the three groups were not statistically significant. Conclusion: Pupillary dilatation with a single drop of 1.0% cyclopentolate, 2.5% phenylephrine or 1% tropicamide is not associated with shallowing of the anterior chamber or narrowing of the ACA in patients with XFS who present with open angles. © 2013 Spanish General Council of Optometry.