The problem of the absence of centrioles in cells of the 1182-4 line of Drosophila melanogaster has been reexamined with high voltage electron microscopy. A hypotonic treatment of the cells before fixation allowed a clear recognition of centrioles in 1 micron thick sections. Three different approaches were used to determine the presence of absence of centrioles in a Kc control cell line and in the 1182-4D cell line. 1) 1500 random 0.5 micron thick sections representing the equivalent of 60 whole cells of the 1182-4D line show no centrioles. In contrast, nearly all centrioles of the Kc cells were detected by this examination. 2) In a blind test 10 grids with either Kc or 1182-4D cells were correctly identified by the operator. In Kc cells, 4 to 6 diplosomes were observed by grid square on about 300 cell profiles, while no centrioles were seen in the sections of 1182-4D cells. 3) Complete serial sections 1 micron thick of whole 1182-4D cells were screened for presence or absence of centriole. No centriole was seen in any section. We conclude that these Drosophila 1182-4D cells, which have been maintained in culture for several years, are free of centrioles.