Fig. 3

Cell migration and invasion can be explored by seeding GFP+ cells onto CAM. Cells re-suspended in 1 mg/ml collagen were seeded onto the surface of live CAM. a, bright field (BV = blood vessel) and b, c, epifluorescence images of live CAM with cells seeded over the surface (Leica MZFLiii stereo microscope with DC500 camera and × 1 Leica lens with × 10 zoom, at room temperature, scale bars = 1 mm). d, HT1080 GFP+ cells dispersed over fixed and stained whole CAM, scale bar = 100 μm. e, f Invaded HT1080 GFP+ cells show different morphologies in fixed and sectioned CAM. g, h, i: a timeline for the invasion of MCF7 GFP+ cells seeded onto CAM shows that cell invasion was evident 1 day after cell seeding and by day 5, cells could be seen within the vasculature and were well disseminated within CAM tissue (scale bars = 25 μm, image planes marked XY, XZ, YZ). Images D-I were taken using Nikon A1 plus confocal microscope, at room temperature. Image D was taken using a × 20 Plan Apo DIC N2, 0.75 NA lens. Image E-I were taken using a × 60 Plan Apo ∞/0.17 WD 0.13, NA 1.40 lens