While it is relatively easy to identify/discriminate neutrophils/eosinophils from freshly isolated peripheral blood by a combination of forward/side scatter properties, autofluorescence, and CD expression by fluorescent antibody (I favour relative expression of FcR, CD16) - it is more problematic to identify neutrophil lineage in cultured leucocytes, especially differentiating from stem cells in vitro, compounded by autofluorescence associated with in vitro culture. Has anyone out there developed a reliable optimised method for this or other related phenotyping issues on in vitro generated leucocyte lineages? _____________________________ Dr G Robin Barclay Lead Scientist / Honorary Senior Lecturer SNBTS Academic Transfusion Medicine Group John Hughes Bennett Laboratory Western General Hospital, Edinburgh, UK Robin.Barclay@ed.ac.uk
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