102 Figure 4 Cornea nerve fiber length (CNFL) and cornea nerve fiber area (CNFA) boxplots, 25th-75th percentiles, whiskers min to max and median lines. Four groups were defined based on established small fiber neuropathy (SFN), probable SFN, no SFN and healthy controls (HC). A: CNFL measured with CCMetrics. B: CNFL measured with ACCMetrics. C: CNFL measured with NeuronJ. D: nerve fiber area (NFA) measured with NFA FIJI. E: NFA measured with ACCMetrics. CNFL and NFA were unable to distinguish between patients with sarcoidosis with established SFN and without SFN. CNF: = corneal nerve fiber length; est. = established; HC = healthy control; prob.= probable; NFA = nerve fiber area; SFN = small fiber neuropathy Correlation between programs CCMetrics, NeuronJ and ACCMetrics obtained values correlated well (correlation coefficient 0.82-0.86, p<0.0001), as did NFA FIJI and ACCMetrics NFA (correlation coefficient 0.54, p<0.0001). The strongest correlation existed between NeuronJ and CCMetrics (R=0.86), see Table 3. Furthermore, inter-rater agreement was excellent between CCMetrics, NeuronJ and ACCMetrics, and fair between NFA FIJI and ACCMetrics. Table 3 Correlation between CCMetrics, ACCMetrics, NeuronJ and NFA FIJI. R-values, p-values and ICC 95%- confidence intervals are presented. Analysis systems R-value P-value ICC 95%-confidence interval CCMetrics vs. ACCMetrics 0.82 <0.0001 0.75-0.87 CCMetrics vs. NeuronJ 0.86 <0.0001 0.80-0.90 ACCMetrics vs. NeuronJ 0.82 <0.0001 0.74-0.87 NFA FIJI vs ACCMetrics NFA 0.54 <0.0001 0.39-0.66 Bland Altman plots comparing the manual analysis method with the semi-automatic and automatic analysis methods were shown in Figure 5, as well as the comparison of NFA FIJI and ACCMetrics NFA. The difference between CCMetrics and ACCMetrics showed a slight ascending slope (Figure 5A), indicating that obtaining mean CNFL with CCMetrics resulted in slightly higher values compared to assessing CNFL with ACCMetrics. A nearly horizontal regression line was seen in the difference between CCMetrics and NeuronJ (Figure 5B), suggesting similar results. There was a slight descending slope in the difference between ACCMetrics and NeuronJ (Figure 5C), indicating that CNFL obtained with ACCMetrics showed slightly lower values than CNFL obtained with NeuronJ. Again, a nearly 6 107 6
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