Indirect immunofluorescence studies have localized fibronectin (FN) within the trunco-conal ridges of the chick embryo heart during the formation of the cushion tissue mesenchyme. Prior to cell migration into the endocardial pads, fluorescence for FN is demonstrated almost entirely in association with the basal surfaces of endocardium and myocardium. Scattered spots and thin dotted-strands of fluorescent material can be demonstrated in the cardiac jelly. Cushion tissue (CT) cells migrating into the cardiac jelly have patches of fluorescent material associated with their surfaces. Filopodial processes always show intense fluorescence. The close association between the fluorescence and the surface of the CT cells suggests that FN may be implicated in the interaction of these cells with the matrical components of the cardiac jelly and, therefore, in the process of cell migration into the endocardial pads. The intensity and amount of FN staining decreased concomitantly with the progressive accumulation of cells in the cushion areas. After the completion of CT cell migration only reduced amounts of faint fluorescence remained in the endocardial pad areas. The possible significance of the changes observed in the distribution of FN during the formation of the cushion tissue mesenchyme is discussed.