Upon contact with the surface of epithelial cells, Shigella flexneri secretes Ipa proteins through the Mxi-Spa type III secretion apparatus. Among the Ipa proteins, IpaB and IpaC form a soluble complex in the bacterial supernatant which appears to be sufficient to initiate the cellular rearrangements necessary to achieve bacterial entry. Here, we provide the first evidence that efficiency of bacterial entry into cells depends on the stage of bacterial growth. Bacteria in the early phase of exponential growth are six times more invasive than those in the stationary phase. The entry efficiency of the bacteria present on the cell surface appears to correlate with the percentage of those that are able to secrete their invasins. This suggests that the capacity to activate the Mxi-Spa apparatus is a major factor in the regulation of bacterial entry efficiency. Consistent with these observations, we have further shown that bacteria which have reached the stage of division secrete Ipa proteins more often than those that have not. Also, initial secretion occurs essentially in the area of the septation furrow. The Ipa proteins, secreted in the vicinity of the septation furrow, seem to initiate the early stages of reorganization of the host cell cytoskeleton.