Title: Analysis and isolation of endocytic vesicles by flow cytometry and sorting: Demonstration of three kinetically distinct compartments involved in fluid-phase endocytosis (endosome/lysosome/cathepsin B/fluorogenic substrates/fluorescein isothiocyanate-dextran)
Abstract: The existence of three distinct classes of endocytic vesicles that are part of the pathway of fluid-phase endocytosis has been demonstrated by flow cytometry. Amounts of fluorescent and scattered light were measured on a particle-by-particle basis for unfractionated whole cell lysates from cells incubated with fluorescein isothiocyanate-dextran. After 20 min two different fluorescent populations were ob- served, and after a 180-min incubation a third highly fluores- cent population was found. Since the fluorescein isothiocya- nate-dextran per fluid-phase vesicle should be a function solely of the external fluorescein isothiocyanate-dextran concentra- tion, the existence of endocytic compartments with widely different amounts of fluorescence could result from a wide range of sizes of initial endocytic vesicles. However, the kinetics of appearance of the intermediate and highly fluorescent vesicles suggest that these compartments become labeled through fusion with the smaller primary endocytic vesicles.
Publication Year: 1985
Publication Date: 1985-01-01
Language: en
Type: article
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