Many effects of insulin, insulin-like growth factors and other receptor stimuli are mediated via the phospholipid second messenger phosphatidylinositol-3,4,5-trisphosphate (PIP(3)). PIP(3) is formed by the activity of phosphoinositide 3-kinases in the plasma membrane, where it serves to recruit signalling proteins. These proteins coordinate complex events leading to changes in cell metabolism, growth, movement and survival. Over the past decade, new techniques for measurements of PIP(3) in the plasma membrane of individual living cells have markedly improved our understanding of the role of this messenger in a variety of cellular processes. This review summarises the mechanisms involved in formation and degradation of PIP(3) in insulin-responsive cells, how PIP(3) can be measured in individual cells as well as accumulating evidence that the plasma membrane PIP(3) concentration undergoes complex spatio-temporal patterns in many types of cells, with particular emphasis on autocrine insulin-induced PIP(3) oscillations in pancreatic beta-cells.