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Chiranjib Majumder,1 Safinaz Mohd Khialdin,2 Subrata Mukhopadhyay,3 Rokiah Omar1 1Optometry & Vision Science Programme, Fakulti Sains Keshihatan, Universiti Kebangsaan Malaysia, Kuala Lumpur, 50300, Malaysia; 2Department of Ophthalmology, Faculty of Medicine, Universiti Kebangsaan Malaysia, Cheras, 56000, Malaysia; 3Shankar Bhaban Netralaya, Rupnarayanpur, Asansol, WB, 713386, IndiaCorrespondence: Rokiah Omar, Email r_omar@ukm.edu.myPurpose: The purpose of this study is to compare the macular thickness, foveal thickness, and retinal nerve fiber layer (RNFL) thickness in amblyopic eyes with those of fellow non-amblyopic eyes using spectral-domain optical coherence tomography (OCT).Methods: A cross-sectional, observational study was conducted with amblyopic subjects from August 2022 to December 2024. A pilot study was carried out using simple random sampling technique. The study involved OCT-based measurement of macular, foveal, and RNFL thickness in amblyopic and non-amblyopic eyes using standardized imaging protocols. Data were analysed using appropriate statistical tests (paired and independent t-tests) based on normality results to compare values across eye types and gender.Results: The results showed significantly higher macular, foveal, and RNFL thickness in amblyopic eyes compared to non-amblyopic eyes (p < 0.05), with no significant differences between anisometropic and mixed amblyopia subtypes. Gender had no significant effect on retinal thickness, except for superior RNFL thickness, which was higher in the anisometropic group of non-amblyopic eyes.Conclusion: This study confirms that amblyopia involves both functional and structural retinal changes, notably in the macula and RNFL, regardless of type or gender. OCT proves valuable in detecting these changes, and future studies should focus on axial length correction and post-treatment outcomes.Keywords: refractive amblyopia, mixed amblyopia, optical coherence tomography, macular thickness, foveal thickness and RNFL thickness