Fig. 4
Uncertainty-guided segmentation ensures efficient mitochondrial quantification in complex skeletal muscle TEM images. (A) Comparison between semi-automatic and probabilistic interactive segmentation results for mitochondria with varying levels of structural clarity. The bottom row shows the uncertainty maps generated by the probabilistic interactive model, where brighter regions indicate areas of higher uncertainty, guiding user attention. (B) Comparison of IoU values derived from semi-automated (SA) and probabilistic interactive (PI) segmentation (whiskers show 10–90 percentile range, N = 6). The arithmetic mean is indicated by a + symbol in each box plot. (C) Fold change comparison of mitochondrial morphological parameters (height, width, area, and count) relative to the gold standard. Multiple parameters were measured for each image. Manual quantification using ImageJ was considered the gold standard, while semi-automatic (SA) and probabilistic interactive (PI) methods produced results via automated quantification based on segmentation masks from each method (mean ± SD, independent two-sample t-test, N = 172–176). (D) Box-and-whisker plot displaying the elapsed time for each method (whiskers represent the 10–90 percentile, p = 1.1 × 10−8, Welch ANOVA followed by Dunnett’s T3 multiple comparisons test, N = 6). The average time for manual segmentation was 2480 s, while the probabilistic interactive method averaged 214 s, representing a 91.4% reduction in analysis time. Statistical significance is indicated by asterisks.
