Fig. 5

dCAM provides a structured 3D environment for studying cell proliferation and migration. a, dCAM partially populated with a co-culture of MDA-MB-231 (white arrows) and MCF7 GFP+ (yellow arrows) breast cancer cells stained with phalloidin for actin cytoskeleton (red) and DAPI nuclear stain (blue). b, MDA-MB-231 cells stained with phalloidin (red) and DAPI (blue) appear to have formed layers over the dCAM surface. c, Cells stained with cell proliferation marker Ki67 (Alexafluor 488, green), phalloidin (red), DAPI (blue) on dCAM. Differential Ki67 staining suggests that not all cells were actively proliferating (proliferating cells – white arrows, high Ki67, low Ki67 cells indicated with yellow arrows). d.1-d.4, Ki67 staining of human cells proliferating and migrating amongst chick CAM cells in invaded CAM (section): combined channels D.1, DAPI, blue (D.2); Phalloidin, red (D.3); Ki67/Alexafluor 647, white (D.4). Images were taken using Nikon A1R confocal microscope operating at room temperature, Image A using a × 20 Plan Apo VC DIC NR, NA 0.75 lens and Images B-D using a × 60 1.40 Plan Apo ∞/0.17 WD 0.13, NA 1.4 lens. Scale bars in C, D = 20 μm. Image planes for 3D images in C and D are marked XY, YZ, XZ