The maintenance of cooperation in populations where public goods are equally accessible to all but inflict a fitness cost on individual producers is a long-standing puzzle of evolutionary biology. An example of such a scenario is the secretion of siderophores by bacteria into their environment to fetch soluble iron. In a planktonic culture, these molecules diffuse rapidly, such that the same concentration is experienced by all bacteria. However, on solid substrates, bacteria form dense and packed colonies that may alter the diffusion dynamics through cell-cell contact interactions. In Pseudomonas aeruginosa microcolonies growing on solid substrate, we found that the concentration of pyoverdine, a secreted iron chelator, is heterogeneous, with a maximum at the center of the colony. We quantitatively explain the formation of this gradient by local exchange between contacting cells rather than by global diffusion of pyoverdine. In addition, we show that this local trafficking modulates the growth rate of individual cells. Taken together, these data provide a physical basis that explains the stability of public goods production in packed colonies.biofilm | evolution | noise | variability | ecology I ron is required for many enzymatic processes, and is therefore an essential nutrient. To overcome the low abundance of free iron in aerobic environments, bacteria, as well as other microorganisms, synthesize and secrete iron-chelating molecules called siderophores (1). Once released in the extracellular environment, siderophores diffuse and complex with iron. Because other bacteria can import them, siderophores can be regarded as public goods (2) and siderophore secretion is often cited as a model system for studying the stability of cooperation (3, 4). In this context, nonproducing mutants, which can enjoy the benefit of siderophores without bearing the cost of their production, have a selective advantage over the WT-producing strain and could, in principle, displace them on evolutionary time scales.This argument usually assumes that public good molecules are readily available to all, as is the case in planktonic cultures, where molecules diffuse freely and rapidly between cells. In liquid conditions, mutants that do not produce siderophores have, in fact, been shown to outcompete WT strains (5-8). However, the limited spatial dispersal of public goods can challenge this picture and has been proposed as a general mechanism for explaining the maintenance of cooperation (8-12). When dispersal is limited, public good molecules tend to stay in the vicinity of the producing subpopulations, allowing them to benefit preferentially from their own production, and thus to balance the advantage of opportunistic nonproducing strains. In many ecological situations, bacteria form complex, tightly packed, and spatially structured colonies, such as biofilms, where public good dispersal may be severely reduced compared with planktonic cultures. This raises the questions of how public good molecules circulate between cells in these natur...