使用流式细胞术测量中性粒细胞吞噬作用的快速全血微量测定法。
Rapid whole-blood microassay using flow cytometry for measuring neutrophil phagocytosis.
作者信息
White-Owen C, Alexander J W, Sramkoski R M, Babcock G F
机构信息
Department of Surgery, University of Cincinnati College of Medicine, Ohio 45267.
出版信息
J Clin Microbiol. 1992 Aug;30(8):2071-6. doi: 10.1128/jcm.30.8.2071-2076.1992.
Abstract
A simple flow cytometric method (FCM) for measuring phagocytosis of Staphylococcus aureus by human neutrophils (polymorphonuclear leukocytes [PMNs]) is described. This assay utilizes 100 microliters of EDTA-anticoagulated whole blood and a simplified method of fluorescently labeling bacteria. A commercially available whole-blood lysing reagent allows for the removal of erythrocytes and the exclusion of external free or adherent bacteria. Phagocytized bacteria are unaffected by this reagent, so PMNs containing internalized bacteria can be easily identified by FCM. Advantages of this method include the following: (i) small sample size, (ii) no requirement for PMN separation, (iii) rapid reliable method of labeling the bacteria, (iv) ability to distinguish between adherent bacteria and those which are actually internalized, (v) avoidance of vital dyes as quenching agents, and (vi) ability to fix cells and store for future FCM analysis.
摘要
本文描述了一种通过人中性粒细胞（多形核白细胞[PMN]）测量金黄色葡萄球菌吞噬作用的简单流式细胞术方法（FCM）。该检测方法使用100微升乙二胺四乙酸抗凝全血以及一种简化的细菌荧光标记方法。市售的全血裂解试剂可去除红细胞并排除外部游离或黏附的细菌。吞噬的细菌不受该试剂影响，因此含有内化细菌的PMN可通过FCM轻松识别。该方法的优点包括：（i）样本量小；（ii）无需分离PMN；（iii）标记细菌的快速可靠方法；（iv）能够区分黏附细菌和实际内化的细菌；（v）避免使用活性染料作为淬灭剂；（vi）能够固定细胞并储存以备将来进行FCM分析。
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Simultaneous measurement of phagocytosis and phagosomal pH by flow cytometry: role of polymorphonuclear neutrophilic leukocyte granules in phagosome acidification.通过流式细胞术同时测量吞噬作用和吞噬体pH值：多形核中性粒细胞颗粒在吞噬体酸化中的作用。
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New, simple flow cytometry technique to discriminate between internalized and membrane-bound particles in phagocytosis.用于区分吞噬作用中内化颗粒和膜结合颗粒的新型简单流式细胞术技术。
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