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Fig. 6 | Respiratory Research

Fig. 6

From: Second harmonic generation microscopy reveals altered collagen microstructure in usual interstitial pneumonia versus healthy lung

Fig. 6

Elastin content and Collagen:Elastin ratio differ in UIP and COP versus healthy lung. a Mature elastin fiber content was similarly decreased in both COP (*p < .007) and UIP (*p < .004) versus healthy lung tissue, and b The total fibrillar collagen/mature elastin ratio was similarly increased in both COP (**p < .004) and UIP (**p < .0008) compared to healthy. Plots represent mean pixel intensity ± SEM for these elastin autofluorescence (Panel a) and (FSHG + BSHG/ Elastin autofluorescence) (Panel b) signals captured as described in Methods, in relative AFU. Plots were generated from a total of 12–24 sections and 60–120 distinct ROIs from N = 4 patients (Healthy and UIP), and 9–18 sections and 45–90 distinct ROIs from N = 3 patients (COP). All data was plotted as N = number of patients per group. Statistics were performed by one way ANOVA with Dunnett’s post-hoc test and correction for multiple comparisons against healthy control. Representative merged images c and d illustrate this lower total FC SHG (blue):mature elastin (green) ratio as seen in c healthy compared to d UIP. (Compare total amount and intensity of the blue summed FSHG + BSHG collagen SHG signal, relative to the green mature elastin signal, in the healthy c versus UIP d panels respectively). Panel d respresents an area of widespread late-stage fibrosis in UIP. For illustrative purposes, the originally grayscale SHG and elastin fluorescence signals are shown with “Blue” and “Green” LUTs applied in ImageJ respectively, with levels (screen stretch) linear and set the same for all channels in all images

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