Harf C, Goffinet S, Meunier O, Monteil H, Colin D A
Laboratoire de Toxinologie Bacterienne, Institut de Bacteriologie de la Faculte de Medecine, Universite Louis Pasteur, Strasbourg, France.
Cytometry. 1997 Mar 1;27(3):269-74.
Endocytosis of fluorescently labelled cells of Legionella pneumophila (L. pneumophila) by free-living Acanthamoeba palestinensis (A. palestinensis) has been studied using flow cytometry. L. pneumophila cells were labelled with CM-DiI, a lipophilic fluorescent probe under conditions that did not modify viability. Coculturing the bacteria with amoebae was accompanied by rapid endocytosis; after 5 min, 90% of the amoebae had internalized bacteria. This percentage remained unchanged during further coculture, but the number of bacteria ingested per amoeba increased. Moreover, the number of ingested bacteria was found to be dependent on the size of the amoeba. The validity of the internalization analyzed by flow cytometry was confirmed by observation using epifluorescence and phase contrast microscopy. CM-DiI labelling associated with flow cytometry provides a very valuable technique for the determination of bacteria endocytosis by free-living amoeba.
利用流式细胞术研究了自由生活的嗜肺军团菌（L. pneumophila）荧光标记细胞被巴勒斯坦棘阿米巴（A. palestinensis）内吞的情况。嗜肺军团菌细胞用CM-DiI（一种亲脂性荧光探针）标记，标记条件不会改变细胞活力。将细菌与变形虫共培养时会迅速发生内吞作用；5分钟后，90%的变形虫内化了细菌。在进一步的共培养过程中，该百分比保持不变，但每个变形虫摄取的细菌数量增加。此外，发现摄取的细菌数量取决于变形虫的大小。通过落射荧光和相差显微镜观察证实了通过流式细胞术分析的内化作用的有效性。与流式细胞术相关的CM-DiI标记为确定自由生活变形虫对细菌的内吞作用提供了一种非常有价值的技术。
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