Fig. 5
Three-dimensional representation of SHG signal in rat Biceps femoris muscle. (a) Left: three-dimensional (3D) reconstruction of SHG collagen fibers in a 350 μm-thick muscle section from 1-year-old wild-type (WT) and dystrophic (DMDmdx) rats (field of view 1.052 × 1.052 × 0.350 mm³). The image was processed with a Laplacian of Gaussian filter to enhance fiber boundaries. Right: individual SHG collagen fibers were detected using the GA3 module in NIS-Elements software (Nikon Europe BV, Amstelveen, Netherlands), with false-coloring applied to the binary mask. (b) Bar graphs illustrate features characterizing endomysial SHG+ objects: density, volume, distance separating objects and elongation of objects. Data were determined in both rat groups considering 3 animals per group and 4 regions each. Statistical analysis was performed using the Wilcoxon test for SHG density. For volume, distance, and elongation datasets, data were discretized into classes, and a log-linear model was applied to compare the distribution between groups. *0.01 < p < 0.05, ***p < 0.001, ****p < 0.0001.
