AbstractThe distribution of forces exerted by migrating Dictyostelium amebae at different developmental stages was measured using traction force microscopy. By using very soft polyacrylamide substrates with a high fluorescent bead density, we could measure stresses as small as 30 Pa. Remarkable differences exist both in term of the magnitude and distribution of forces in the course of development. In the vegetative state, cells present cyclic changes in term of speed and shape between an elongated form and a more rounded one. The forces are larger in this first state, especially when they are symmetrically distributed at the front and rear edge of the cell. Elongated vegetative cells can also present a front‐rear asymmetric force distribution with the largest forces in the crescent‐shaped rear of the cell (uropod). Pre‐aggregating cells, once polarized, only present this last kind of asymmetric distribution with the largest forces in the uropod. Except for speed, no cycle is observed. Neither the force distribution of pre‐aggregating cells nor their overall magnitude are modified during chemotaxis, the later being similar to the one of vegetative cells (F0 ∼ 6 nN). On the contrary, both the force distribution and overall magnitude is modified for the fast moving aggregating cells. In particular, these highly elongated cells exert lower forces (F0 ∼ 3 nN). The location of the largest forces in the various stages of the development is consistent with the myosin II localization described in the literature for Dictyostelium (Yumura et al.,1984. J Cell Biol 99:894–899) and is confirmed by preliminary experiments using a GFP‐myosin Dictyostelium strain. Cell Motil. Cytoskeleton, 2008. © 2008 Wiley‐Liss, Inc.