Background: This study assessed the impact of contact lens wear on retinal spectral domain optical coherence tomography (SD-OCT) image quality and macular thickness measurements, among subjects with myopia. Methods: This was a prospective study including 34 subjects (26.59 AE 3.19 years) with myopia or myopic astigmatism. Twelve were imaged wearing spherical soft contact lenses, eight non-contact lens wearers were imaged with a plano soft contact lens, and 14 with significant astigmatism were fitted with a rigid gas permeable (RGP) contact lens. For each group of contact lens types, the average image quality index (Q-index), and the average macular thickness measurements were compared between macular OCT scans obtained from the same eyes with and without a contact lens. Results: Among the subjects assessed with their habitual spherical soft lenses, the average Q-index was similar for scans acquired with and without a contact lens (30.10 AE 1.94 versus 31.03 AE 2.55; p = 0.18). Among non-contact lens wearers, the average Q-index was slightly higher for scans acquired without a contact lens, compared to scans with a plano contact lens (31.99 AE 2.06 versus 29.51 AE 1.56; p = 0.006). Among 14 subjects imaged wearing a fitted RGP contact lens, the Q-index was similar for scans acquired with and without a contact lens (29.04 AE 2.73 versus 28.75 AE 2.86; p = 0.78). In all groups, there were no correlations between the power of the sphere and change in the Q-index (that is, post-minus pre-contact lens Q-index), and no differences were found between OCT-derived macular thickness measurements from scans with and without a contact lens. The magnitude of cylinder was not correlated with the change in the Q-index in the habitual and RGP contact lens groups. However, an inverse correlation between cylinder power and change in the Qindex was found in the plano contact lens group. Conclusion: In low to intermediate levels of myopia, with or without regular astigmatism, macular SD-OCT imaging does not merit placement of a soft or rigid contact lens, nor is there an added benefit from removing a habitual spherical soft lens prior to scanning.