1. 3D model and in vivo live imaging of dermal wound repair
3D model and in vivo live imaging of dermal wound repair A(x,y) Cross‐section of the computational simulation of wound healing in adult dermis. Please refer to associated Movie EV4.B–DLive imaging of adult PDGFRαH2BEGFP back skin during wound healing. (B) Experimental design of in vivo imaging of wound healing. (C) Representative wound bed Z‐stack stitched maximum projections of the entire wound bed (upper panel) showing fibroblasts (green) and collagen with second harmonic generation (SHG) in purple at indicated time points after wounding and before live imaging. Dotted line indicates wound edge, and boxed area indicates the position of the magnified area below. (D) Representative time‐lapse images of adult wound bed at PW4, where Z‐directional movement is visualised by pseudo‐colouring the fibroblast nuclei (red, movement towards the basement membrane; blue, movement away from the basement membrane). Dotted line indicates wound edge. Please refer to associated Movie EV6 which shows fibroblasts actively migrating into the wound bed.E–HIn vivo lineage tracing of upper (Blimp1Cre) and lower (Dlk1CreER) dermal fibroblasts during wound healing. (E) Experimental strategy for in vivo lineage tracing. (F) Immunofluorescence image of Blimp1Cre × Confetti (upper panels) and Dlk1CreER × Confetti (lower panel) back skin inside and outside the wound bed at 10 days postwounding. Colours show YFP (green), RFP (red) and CFP (blue) with bright field image overlaid. (G) Clone size quantification of Blimp1Cre‐labelled fibroblasts inside and outside the wound bed at 10 days postwounding (n = 400 clones of four biological replicates). (H) Clone size quantification of Dlk1CreER‐labelled fibroblasts inside and outside the wound bed at 10 days postwounding (n = 300 clones of two biological replicates).Data information: Scale bars, 500 μm (C); 100 μm (D, F).
Emanuel Rognoni et al. Mol Syst Biol 2018;14:e8174
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