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A methodology for detection of phagocytosis of murine neutrophilsby by flow cytometry

ZHAO Xiu chun,YAO Chun yan,LI Bai qing,XU Feng zhen (Department of Respiratory Disease,Affliated Hospital Bengbu Medical College,Anhui 233004,China)  
Objective:To establish the method for detection of phagocytosis of muirne neutrophils by flow cytometry.Methods: Staphylococcus aureus ( S.aureus ) and Escherichia coli ( E.coli ) were labeled with fluorescein isothiocyanate(FITC) in different buffers.The efficient rate of labeling was assessed by flow cytometry.Whole blood taken from mice were incubated with FITC labeled bacteria at 37℃ for 10-20 min,followed by lysing red blood cells.The fluorescent intensity of neutrophils was measured as phagocytotic rate by flow cytometry.Results:The good labeling of S.aureus and E.coli were obtained in PBS(pH 7.2) and carbonate buffer(pH 9),respectively.The percentage of phagocytosis of E.coli was higher than that of S.aureus by murine neutrophils.Conclusions:Detection of phagocytosis of FITC labeled E.coli by flow cytometry is simple,rapid and reproducible method for measuring phagocytotic activity of murine neutrophils.
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