2001•Unpublished venueRequires access
A Flow Cytometric Assay for Quantitating Phagocytic and Oxidative Burst Activity of Bovine Polymorphonuclear Leukocytes in Whole Blood
HU Song-hua
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Abstract
A rapid and sensitive flow cytometric method for simultaneous measurement of phagocytosis rate and oxidative burst activity of bovine polymorphonuclear leukocytes(PMN) in small amount of blood samples is described.After incubation of the PMN with 1.75 μm blue fluorescent beads,intracellular dihydrorhodamine 123(DHR) was oxidized into green fluorescent rhodamine 123(ROD) by oxidative burst products.Cells with beads and ROD were simultaneously detected by a flow cytometer.The method also allows to count the number of beads ingested by PMN.Phagocytosis and oxidative burst activity both increased with increasing beads concentration.A ratio of 50 beads to one cell was considered optimal for assaying bovine blood PMN activity.To evaluate the reliability of the method,blood samples were treated with a phagocytosis inhibitor,cytochalasin B.Incubation of blood with cytochalasin B at 1,2.5 or 5 mg/L resulted in a decrease in the phagocytosis rate with a concommitant decrease in oxidative burst activity.