Distribution of acridine orange fluorescence in Plasmodium falciparum-infected erythrocytes and its implications for the evaluation of digestive vacuole pH
Distribution of acridine orange fluorescence in Plasmodium falciparum-infected erythrocytes and its implications for the evaluation of digestive vacuole pH
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DOI：
10.1016/s0166-6851(01)00403-0
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发表时间：
2002-02-01
影响因子：
1.5
通讯作者：
Ward, SA
中科院分区：
文献类型：
--
作者：
Bray, PG;Saliba, KJ;Ward, SA
P.用吖啶橙子（AO）染色法对恶性疟原虫K1株与P.恶性疟原虫，其中消化泡用膜不渗透的葡聚糖连接的荧光标记物特异性标记。AO荧光仅限于寄生虫质膜内的区域。AO染色的寄生虫内的暗区与用葡聚糖-SNARF标记的区域和在明视野图像中观察到的暗区完全一致。该区域代表整个消化泡。与AO获得的图像相似的图像也获得了细胞溶质pH指示剂BCECF，作为膜渗透剂乙酰氧基甲酯加载到寄生的红细胞中。与AO染色的寄生虫一样，BCECF负载的细胞显示来自寄生虫胞质溶胶的强荧光，而来自寄生虫消化泡的荧光不存在。使用皂苷从其宿主细胞分离的寄生虫的实验得到了非常相似的结果与完整的寄生红细胞：AO和BCECF都标记寄生虫胞质溶胶，没有荧光从消化泡。
The pattern of fluorescence fromP. falciparumK1 strain stained with acridine orange (AO) was compared with that fromP. falciparumin which the digestive vacuole was specifically labelled with membrane-impermeant dextran-linked fluorescent markers. AO fluorescence is restricted to the area within the parasite plasma membrane. The dark region within the AO-stained parasite coincides exactly with the region labelled with dextran-SNARF and with the dark region observed in the brightfield image. This region represents the digestive vacuole in its entirety. Similar images to those obtained with AO were also obtained with the cytosolic pH indicator BCECF, loaded into parasitized erythrocytes as the membrane-permeant acetoxymethyl ester. As with AO-stained parasites, BCECF-loaded cells showed strong fluorescence from the parasite cytosol, with no fluorescence from the parasite's digestive vacuole. Experiments with parasites isolated from their host cells using saponin gave very similar results to those obtained with intact parasitized erythrocytes: AO and BCECF both labelled the parasite cytosol, with no fluorescence from the digestive vacuole.