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Fig. 3. The distance between neighbouring podosomes depends on the substrate flexibility. (A) Total internal fluorescence microscopy of a podosome cluster. Podosomes are hardly visible over the dense actin cloud. (B) Image post-processing by a top-hat filter reveals the location of podosomes. Bars, 10 µm. (C) Computation of the mean distance between nearest podosomes for 16 pairs of podosomes per cell over full time-lapse sequences and for each substrate rigidity (n=96 for 0.08% gel; n=112 for 0.2% gel; and n=144 for glass). Data are expressed as mean ± s.d.