Fluorescence-activated cell sorting (FACS) of Drosophila hemocytes reveals important functional similarities to mammalian leukocytes

摘要: Drosophila is a powerful model for molecular studies of hematopoiesis and innate immunity. However, its use functional cellular remains hampered by the lack single-cell assays hemocytes (blood cells). Here we introduce generic method combining fluorescence-activated cell sorting nonantibody probes that enables selective gating live from lymph glands (larval hematopoietic organ) or hemolymph equivalent). Gated are analyzed sorted at will based on precise quantitation fluorescence levels originating metabolic indicators, lectins, reporters (GFP β-galactosidase) antibodies. With this approach, discriminate sort plasmatocytes, major hemocyte subset, lamellocytes, an activated subset present in gain-of-function mutants Janus kinase Toll pathways. We also illustrate how important, evolutionarily conserved, blood-cell-regulatory molecules, such as calcium glutathione, can be studied functionally within hemocytes. Finally, report vivo transfer their successful reanalysis retrieval single hosts. This versatile approach detection, analysis, sorting, which efficient down to one animal, should critically enhance ex other species, notably mosquitoes.