Surgical treatments of macular holes have become increasingly effective in inducing resolution of the cuff of surrounding subretinal fluid, resulting in increased vision in many patients. However, for many conditions that mimic a macular hole, either surgery is not indicated or different surgical manipulations are necessary. Differentiating macular holes from some forms of macular pseudoholes can be difficult or impossible based solely on clinical examination. Adjunctive tests that may enhance the accuracy of diagnosis are either not feasible or not available to most clinical practices. We evaluated three clinic-based tests for their value in allowing the differentiation between macular holes and macular pseudoholes: Amsler grid testing, Watzke-Allen sign, and laser aiming beam test. These tests were evaluated in three groups of clinically defined patients: those with full-thickness macular holes, those with macular pseudoholes, and those who had previously undergone successful macular hole treatment. Although the Amsler grid testing was sensitive in correlating with clinically defined macular holes, it was not specific. The Watzke-Allen sign and, to a greater extent, the laser aiming beam test were extremely sensitive and specific in correlating clinically defined full-thickness macular holes and pseudoholes. These tests improve the accuracy of diagnosis of full-thickness macular holes.