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流式细胞术检测小鼠中性粒细胞吞噬功能的方法学探讨

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

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【作者】 赵修春; 姚春艳; 李柏青; 徐凤珍;

【Author】 ZHAO Xiu chun,YAO Chun yan,LI Bai qing,XU Feng zhen (Department of Respiratory Disease,Affliated Hospital Bengbu Medical College,Anhui 233004,China)

【机构】 蚌埠医学院附属医院呼吸内科; 蚌埠医学院免疫学教研室; 蚌埠医学院附属医院呼吸内科 (现工作单位:广东省广州市卫生学校; 510000); 安徽蚌埠233004; 安徽蚌埠233003; (现工作单位:广东省广州市卫生学校; 安徽蚌埠233004;

【摘要】 目的 :建立用流式细胞术检测小鼠中性粒细胞 (PMN)吞噬功能的方法。方法 :用不同缓冲液制备荧光素 (FITC)标记葡萄球菌或大肠埃希菌 ,流式细胞术检测细菌FITC标记率。小鼠全血与FITC标记细菌 37℃共育 1 0~ 2 0min ,溶解红细胞后用流式细胞术检测小鼠PMN荧光强度的变化 ,计算PMN吞噬率。结果 :用PBS(pH 7.2 )标记金黄色葡萄球菌效果好 ,用碳酸盐缓冲液 (pH 9)标记大肠埃希菌效果好。小鼠PMN对金黄色葡萄球菌的吞噬率低 ,对大肠埃希菌的吞噬率高。 结论 :用流式细胞术检测小鼠PMN吞噬FITC标记大肠埃希菌是测定小鼠PMN吞噬功能的简便、快速和重复性好的方法

【Abstract】 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.

  • 【文献出处】 蚌埠医学院学报 ,Journal of Bengbu Medical College , 编辑部邮箱 ,2004年05期
  • 【分类号】R392
  • 【被引频次】22
  • 【下载频次】894
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