Abstract Cytoplasm from lactating cells is included with some milk fat globules at secretion. The objective was to search for factors causing this phenomenon. Globules bearing crescents of cytoplasm were selectively stained with the fluorescent dye, acridine orange, and their proportion in the globule population was obtained by counting from photomicrographs. Incidence of crescents on fat globules in milk samples of 50 human donors ranged from 1 to 29%, and the mean was 7.2%. Two bovine milk samples, both representing over 100 animals, contained 1% or less of globules with crescents. Globules in individual milkings of five beef cows showed the same low proportion of crescents. In addition to species, genetic and diurnal factors Influenced numbers of crescents. Two sisters showed evidence within and between lactations of a persistent high proportion (>25%) of globules with crescents. Samples collected in the a.m. contained a lower percentage of globules with crescents (6.5%) than those obtained in the p.m. (9.7%). Crescent incidence was not correlated with lipid or protein content of human milk, interval within a milking, days in lactation, or the donor's age. Evidence is presented to suggest that the concentration, distribution, and acylation of butyrophilin and xanthine oxidase, coat proteins of the apical plasma membrane, are important factors in globule crescent formation.