Filipin binding to membrane sterols induces deformations of the membrane that are detected by freeze-fracture either as 20- to 25-nanometer protuberances or as pits on the fracture faces. By using the filipin probe in pancreatic acinar cells, it was found that the polarity of filipin-induced deformations in the membrane limiting the Golgi condensing vacuoles is opposite that in the membrane limiting the mature zymogen granules. This asymmetry could be due to unequal partitioning of cholesterol between the membrane leaflets in these two compartments during the transformation of the condensing vacuole into the zymogen granule.