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BACKGROUND Photographs or videos taken from a mobile phone with a +20-D lens can be an alternative to indirect ophthalmoscopy in mass screening and telemedicine. OBJECTIVE To develop a mobile fundus camera (MFC) that ensures eye screening with safety, ease of use, and high accuracy in detecting fundus lesions. METHODS From October 1st, 2021, to September 30th, 2022, we developed an MFC based on Samsung S21 and iPhone 12 mobile phones with four different types of connecting lenses. Ten ophthalmology personnel were trained to use the MFC on schematic eyes and evaluate the developed systems. The four schematic eyes had a normal fundus, diabetic retinopathy (DR), normal optic disc, and glaucomatous optic disc. The system safety was assessed based on the ISO 15004-2 Visible wavelength (400–700 nm). All users completed the Thai version of the Usability Experience Questionnaire (UEQ). The evaluation of image quality was based on the sharpness index (SI), disc-to-image ratio, and color discrimination, which included relative redness, red-green difference, and red-blue difference. RESULTS Our system satisfied the safety criteria. The mean UEQ scores for the overall MFCs were above average in the domains of attractiveness, dependability, stimulation, and novelty. However, the efficiency and perspicuity were below average. The MFC based on the Samsung S21 had a significantly better overall SI but poorer overall color discrimination power than that of the iPhone 12 (p < 0.05). The best SI was achieved using Samsung S21+Volk 28D, whereas iPhone 12+oDocs 20D provided the best color discrimination. Interparticipant differences were not statistically significant. CONCLUSIONS All the developed MFCs tested were safe and usable. Based on the SI criteria, the Samsung S21+ Volk 28D should be used for further development.