After microinjection into cultured Vero cells, rhodamine‐labeled clathrin triskelions gave rise to a punctuate fluorescence pattern typical for clathrin, with two major localizations: the plasma membrane and the perinuclear region of the cells. We analyzed clathrin motion by Fluorescence Recovery After Photobleaching and its 3 dimensional distribution by Confocal Microscopy. Altogether, 55% of total clathrin is polymerized into coats which are turning over with a half time of 11 seconds and 45% of total diffuses freely in the cytoplasm. Various conditions known to affect membrane traffic were investigated. Cytosolic acidification or ATP depletion stabilized the polymerized clathrin coats without modifying the ratio of free versus polymerized clathrin. Low temperature (6 °C) or hypertonic media dramatically increased both the stability and the amount of the polymerized clathrin. We conclude that ATP and pH homeostasis are needed to support a very high turnover of the clathrin coats in living cells whereas low temperature and high osmotic strength promote an extensive polymerization of clathrin.