Purpose: To determine the status of sub-specialization among Nigerian ophthalmologists as well as their dispositions and barriers against sub-specialization with a view to providing valuable information for the purpose of human resources for eye care planning thereby providing useful insight into the future of ophthalmic practice in Nigeria. Methods: This was a web-based, cross-sectional study conducted among Ophthalmologists in Nigeria. An online questionnaire was distributed through e-mails using Qualtrics software (Qualtrics, Provo, UT, USA). Information concerning socio-demographic characteristics, type of practice, location of practice, years of practice, status and disposition to sub-specialization as well as barriers to sub-specialization were obtained through the questionnaire.Results: two hundred and four Nigerian Ophthalmologists participated in the study out of which 118 (57.8%) were females. One hundred and ten (54.0%) respondents had undergone sub-specialty training. The sub-specialties with the highest number of patronage was Paediatric Ophthalmology and Strabismus (14.2%). Respondents older than 46 years were three times more likely to have undergone subspecialty training compared to respondents who were aged 46 years and below [odds ratio (OR) = 3.01, 95% Confidence interval (CI) = 1.33 – 6.83, p = 0.01]. The main barriers to the availability and uptake of sub-specialty services as well as the challenges of sub-specialty services at the centres with established sub-specialty practice were non-availability/inadequate trained specialist and inadequate equipment. Conclusion: Nigerian Ophthalmologists are well disposed to sub-specialization although the extent of sub-specialization among them was a little above average. The main barriers to the availability and uptake of sub-specialty services as well as the challenges of sub-specialty services at the centres with established sub-specialty practice in this study were non-availability/inadequate trained specialist and inadequate equipment.