Cantinieaux B, Hariga C, Courtoy P, Hupin J, Fondu P
Department of Haematology, University Hospital St. Pierre, Brussels, Belgium.
J Immunol Methods. 1989 Jul 26;121(2):203-8. doi: 10.1016/0022-1759(89)90161-0.
A new method for measuring the uptake of Staphylococcus aureus by human polymorphonuclear neutrophils (PMN) using flow cytometry (FCM) is described. Bacteria were labeled with fluorescein isothiocyanate (FITC) and incubated with PMN in a suspension assay. At the end of the assay, phagocytosis was arrested by the addition of cold paraformaldehyde. Thereafter, phagocytosis was quantitated by FCM, using crystal violet to distinguish adherent versus ingested bacteria. The method was validated in multiple samples by reference to our microscopic technique; correlations of phagocytic indices were in good agreement. The FCM method was also found to be reproducible and provided a mean to detect low phagocyte values. Another feature of the approach is that FCM readings can be delayed without any appreciable alterations in the results.
描述了一种使用流式细胞术（FCM）测量人多形核中性粒细胞（PMN）摄取金黄色葡萄球菌的新方法。细菌用异硫氰酸荧光素（FITC）标记，并在悬浮试验中与PMN一起孵育。试验结束时，通过加入冷多聚甲醛来阻止吞噬作用。此后，使用结晶紫区分粘附细菌与摄入细菌，通过FCM对吞噬作用进行定量。通过参考我们的显微镜技术，该方法在多个样本中得到验证；吞噬指数的相关性非常一致。还发现FCM方法具有可重复性，并提供了一种检测低吞噬细胞值的手段。该方法的另一个特点是FCM读数可以延迟，而结果没有任何明显变化。
