AIM: To compare central macular thickness (CMT) measurements obtained by two spectral-domain optical coherence tomography (SD-OCT) exams, and to evaluate measurement reproducibility and agreement between these two exams, and to investigate the relationship between CMT and possible influencing factors such as age, sex, eye (OD/OS), and operators in elderly non-mydriatic eyes. METHODS: Seventy-two normal subjects were included. Every subject underwent CMT measurement twice using one of two SD-OCT (OSE-2000, Moptim, Shenzhen, China & 3-D OCT-1000, Topcon, Tokyo, Japan) instruments respectively where we randomly chose one eye in each patient for the test; these exams were performed by two operators over an hour period with a brief rest between sessions. Comparison of the OSE-2000 and 3-D OCT-1000 CMT measurements was based on paired-t test. The mean difference between the CMT measurements was calculated. General linear model analyzed the relationships among eye (OD/OS), operator, sex, and CMT values using age as co-variant. All tests were considered statistically significant at P<0.05. The main outcome measures included CMT. RESULTS: When evaluated with general linear model analysis, CMT measurements were found to have high reproducibility across the two instruments between the two operators for the OSE-2000 single line scan and 3-D OCT-1000 macular scans (P=0.731; P=0.443). There was statistically significant difference in CMT values between the two instruments (P<0.001) and the mean difference was -46.83μm at 95% confidence limits (-49.15,-44.51). Age was positively correlated with CMT (beta coefficient = 0.516, P=0.001; beta coefficient = 0.453, P=0.009) and sex was correlated with CMT from the OSE-2000 (P=0.021) but not with the 3-D OCT-1000 (P=0.056). According to the actual thickness measurements, the CMT of the male was thicker than the female’s but there was no statistical difference. There was interaction between sex and eye in OSE-2000 and not in 3-D OCT-1000 (P=0.02; P=0.374). No significant correlation was found between CMT and the influencing factor of eye in both of the instruments (P=0.884; P=0.492). CONCLUSION: Reproducibility of CMT measurement using the two SD-OCTs is excellent in normal eyes according to the operator factor analysis. OSE-2000 has a different posterior retinal boundary of CMT measurement, which results in the CMT value differences, compared with the 3-D OCT-1000. Age is positively correlated with CMT measurement while sex is correlated with CMT in the OSE-2000 but not in the 3-D OCT-1000 and eye (OD/OS) had no correlation with CMT values. Mydriatic drops may not be necessary for CMT measurement using high scan rate SD-OCT in normal eyes in dark room.