Simultaneous measurement by flow cytometry of phagocytosis and hydrogen peroxide production of neutrophils in whole blood

Bacterial ingestion and hydrogen peroxide production by polymorphonuclear leukocytes (PMN) were simultaneously measured using flow cytometry and 2′,7′-dichlorofluorescein diacetate and fluorescent Staphylococcus aureus. Only 100 μ1 of a whole blood specimen were required for these determinations, and the results were found to be independent of the absolute numbers of PMN, making the purification and adjustment of PMN numbers unnecessary. A positive correlation between phagocytosis and hydrogen peroxide production by individual PMN was demonstrated in healthy adults.