Computational protocol: Effects of photocoagulation on ocular blood flow in patients with severe non-proliferative diabetic retinopathy

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Protocol publication

[…] Binarization of the choroidal area in EDI-OCT images was performed by the modified Niblack’s method as previously reported.[] EDI-OCT images were analyzed using the ImageJ software (ImageJ version 1.47, NIH, Bethesda, MD, USA). The examined area was 1,500 μm wide in the subfoveal choroid, extending vertically from the RPE to the chorioscleral border (). This choroidal area was selected using the ImageJ ROI Manager. Next, the image was converted into 8 bits. The vitreous cavity in front of the macular area was selected by the Oval Selection Tool on the ImageJ tool bar, and the maximum reflectivity of these areas was determined. The maximum brightness was set at the minimum value to minimize noise in the OCT image. After adjusting by the Niblack Auto Local Threshold, the luminal area was determined using the threshold tool. The light pixels were defined as the stromal areas, and the dark pixels were defined as the luminal areas. After adding the data of the distance of each pixel, the luminal and stromal areas were automatically calculated. Two clinicians blinded to the other findings measured the area. [...] The value of each parameter was presented as the mean ± standard deviation. For comparisons of categorical data, the chi-square test was used, whereas quantitative data were compared using Kruskal–Wallis tests. Analysis of variance with post-hoc Bonferroni correction was used to evaluate data pertaining to blood flow parameters and choroidal areas. Spearman’s rank correlation coefficient tests were used to determine the correlation coefficients between the variables. Multiple stepwise regression analysis was used to determine the association between blood flow parameters and other variables. All statistical analyses were performed using IBM SPSS Statistics for Windows, v.23 (IBM Corp., Armonk, NY). The significance level was set at a probability (p) value < 0.05. […]

Pipeline specifications

Software tools ImageJ, SPSS
Applications Miscellaneous, Microscopic phenotype analysis
Organisms Homo sapiens
Diseases Diabetes Mellitus