Fig. 4

dCAM provides a collagen-rich 3D substrate for cell culture. Laser scanning confocal spectral unmixing was used to determine the residual components after decellularization of CAM (Nikon A1 Plus at room temperature using a × 60 1.40 Plan Apo ∞/0.17 WD 0.13, NA 1.4 lens). a, combined image, b DAPI only, c CAM background only, d, phalloidin for cellular actin cytoskeleton (scale bar for A = 20 μm). Scanning electron microscopy (SEM) was used to characterize the surface of the decellularized tissue (Quanta FEI), e and f show dCAM surface features including vasculature (yellow arrows) and fibrous extracellular matrix (scale bars: E = 50 μm, F = 5 μm). dCAM used as a growth matrix: g shows a bright field image of dCAM during colonization and h shows MDA-MB-231 GFP+ cells adhering and proliferating over the dCAM (DC, yellow arrows). Images G and H were taken using a Zeiss Axio Vert inverted epifluorescence microscope and × 5 Planar Plan Neofl Ph1 0.15 ∞ /0.17 lens with the DS-Fi2 camera, operating at room temperature, scale bars = 1 mm