Fig. 4

Statistical validation of MuSeeQ. a–c Technical reproducibility assays for the measurement of the area of the seeds (a), the area of released soluble mucilage (b) and the ratio (AreaM/AreaS) (c) on seeds from the cultivar Oliver sown on three different agarose stained gels. ns indicate no significant differences (p ≤ 0.05) as determined by one-way ANOVA followed by a Tukey’s HSD (honest significant difference) post hoc test (n = 30 seeds for each technical replicat). Bars represent mean ± SEM d Correlation analysis of automated (MuSeeQ) versus manual (Fiji) measurements of the ratio (AreaM/AreaS) (n = 104 seeds from 21 randomly selected flax RILs, 5 seeds each). e A representative result of segmentation using MuSeeQ and SmartGrain on the same seed. Note that both image analysis tools numbered the seed 14. Opposite to SmartGrain, MuSeeQ does not draw the computed parameters on the seed, except for the seed outlines. f Bland–Altman plot showing no major statistical differences between the results of AreaS obtained using both methods. g Correlation analysis of automated (MuSeeQ) versus manual (SmartGrain) measurements of the AreaS. Pearson’s correlation coefficients were determined by linear regression analyses (n = 104 seeds from 21 randomly selected flax RILs, 5 seeds each). Red lines are least squares regression curves. Bars: 1.5 mm