Conventional static automated perimetry provides important clinical information, but its utility is limited by considerable test-retest variability. Fixational eye movements during testing could contribute to variability. To assess this possibility, it is important to know how much sensitivity change would be caused by a given eye movement. To investigate this, we have evaluated the gradient, the rate at which sensitivity changes with location. We tested one eye each, twice within 3 weeks, of 29 patients with glaucoma, 17 young normal subjects and 13 older normal subjects. The 10-2 test pattern with the SITA Standard algorithm was used to assess sensitivity at locations with 2 degrees spacing. Variability and gradient were calculated at individual test locations. Matrix correlations were determined between variability and gradient, and were substantial for the patients with glaucoma. The results were consistent with a substantial contribution to test-retest variability from small fixational eye movements interacting with visual field gradient. Successful characterization of the gradient of sensitivity appears to require sampling at relatively close spacing, as in the 10-2 test pattern.