Purpose:To develop semi-automated application software that quickly analyzes infrared meibography images taken with the CSO Sirius Topographer (CSO, Italy) and to compare them to the manual analysis system on the device (Phoenix software platform).Methods:A total of 52 meibography images verified as high quality were used and analyzed through manual and semi-automated meibomian gland (MG) detector software in this study. For the manual method, an experienced researcher circumscribed the MGs by putting dots around grape-like clusters in a predetermined rectangular area, and Phoenix software measured the MG loss area by percentage, which took around 10 to 15 minutes. MG loss was graded from 1 (<25%) to 4 (severe >75%). For the semi-automated method, 2 blind physicians (I and II) determined the area to be masked by putting 5 to 6 dots on the raw images and measured the MG loss area using the newly developed semi-automated MG detector application software in less than 1 minute. Semi-automated measurements were repeated 3 times on different days, and the results were evaluated using paired-sample t test, Bland-Altman, and kappa analysis.Results:The mean MG loss area was 37.24% with the manual analysis and 40.09%, 37.89%, and 40.08% in the first, second, and third runs with the semi-automated analysis (P < 0.05). Manual analysis scores showed a remarkable correlation with the semi-automated analysis performed by 2 operators (r = 0.950 and r = 0.959, respectively) (P < 0.001). According to Bland-Altman analysis, the 95% limits of agreement between manual analysis and semi-automated analysis by operator I were between-10.69% and 5% [concordance correlation coefficient (CCC) = 0.912] and between-9.97% and 4.3% (CCC = 0.923) for operator II. The limit of interoperator agreement in semi-automated analysis was between-4.89% and 4.92% (CCC = 0.973). There was good to very good agreement in grading between manual and semi-automated analysis results (0.76-0.84) and very good interoperator agreement with semi-automated software (0.91) (P < 0.001).Conclusions:For the manual analysis of meibography images, around one hundred dots have to be put around grape-like clusters to determine the MGs, which makes the process too long and prone to errors. The newly developed semi-automated software is a highly reproducible, practical, and faster method to analyze infrared meibography images with excellent correlation with the manual analysis.