Journal: Scientific Reports
Article Title: Lumican delays melanoma growth in mice and drives tumor molecular assembly as well as response to matrix-targeted TAX2 therapeutic peptide
doi: 10.1038/s41598-017-07043-9
Figure Lengend Snippet: Investigation of the relationship between lumican and intratumoral collagen organization. ( a ) Representative microphotographs of s.c. allograft sections stained with picrosirius red and viewed under dark-field cross-polar optics (original magnification ×63). Birefringence of collagen fibers allows distinction between type I (red) and type III (green) collagen. ( b , c ) Quantification of the relative distribution of red ( b , left panel ) and green ( c , left panel ) pixels as well as the corresponding intensities ( b and c , right panels ), expressed as a percentage of mean pixel intensity in the Lum +/+ (WT) group (mean ± SEM, t test, * p < 0.05). ( d ) Quantification of type I/type III ratio based on red and green pixel calculations, respectively. Individual ratios were calculated from 3 different fields per animal, and then averaged (mean ± SEM, t test, *** p < 0.001). ( e – g ) Quantification of tumor ECM collagen organization from spectra derived from Gabor filtering and FFT. ( e ) Flowchart depicting the different stages of image analysis. From grey scale-converted picrosirius-stained images, a 3 × 3 median filter is applied before Gabor filtering and then FFT. After determining the elliptical shape of the scatter pattern (see red ellipses on right panel ), measurements of the elliptical axes ( black lines ) generated from ω angle values n 1 to n 4 were performed so as to produce a collagen orientation index (N), according to displayed equations ( inset ). ( f ) Histogram displays results of collagen orientation index determination within melanoma tumors allografted to Lum +/+ or Lum −/− mice (mean ± SEM, t test, ** p < 0.01). ( g ) Correlation between calculated collagen orientation index and final tumor volume in Lum +/+ (WT, black dots ) and Lum −/− ( red dots ) mice. Linear regression was performed ( black line ) and r coefficient arising from non-parametric two-tailed Spearman test was determined. ( h ) Representative collagen SHG images from B16F1 tumors (original magnification ×20). ( i ) Collagen density of SHG images for s.c. allografts of Lum +/+ and Lum −/− animals (mean ± SEM, t test, ** p < 0.01). ( j ) Representative polar plots of SHG intensity vs . angle of laser polarization.
Article Snippet: To assess the basketweave structure of collagen, an innovative bioimaging approach combining Fast Fourier Transform (FFT) with Gabor filtering was applied .
Techniques: Staining, Derivative Assay, Generated, Two Tailed Test