THIAZOLE ORANGE - A NEW DYE FOR PLASMODIUM SPECIES ANALYSIS
THIAZOLE ORANGE - A NEW DYE FOR PLASMODIUM SPECIES ANALYSIS
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DOI：
10.1002/cyto.990080606
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发表时间：
1987-11-01
期刊：
影响因子：
--
通讯作者：
RECKTENWALD, D
中科院分区：
文献类型：
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作者：
MAKLER, MT;LEE, LG;RECKTENWALD, D
提出了一种快速灵敏的体外培养中恶性疟原虫测定方法。该技术采用荧光流式细胞仪，配备可发射 488 nm 光的 15 兆瓦氩激光器和可对 RNA 进行染色的膜渗透性荧光染料噻唑橙 (TO)。将寄生的红细胞悬浮在含有 10-5 M TO 的 1 ml 磷酸盐缓冲盐水 (PBS) 中，并在室温下黑暗中孵育该混合物 15 分钟，对寄生红细胞进行染色。染色的细胞可以新鲜进行分析，也可以在用 1% 多聚甲醛/PBS 或 0.25% 固定后进行分析，然后进行染色。荧光标记的寄生红细胞数量与吉姆萨染色细胞数量之间存在极好的对应关系。
A rapid sensitive method for the determination of Plasmodium falciparum in in vitro culture is presented. The technique employs a fluorescent flow cytometer equipped with a 15-mwatt argon laser that emits light at 488 nm and a membrane-permeable fluorochrome thiazole orange (TO) that stains RNA. Parasitized red cells are stained by suspending them in 1 ml of phosphate-buffered saline (PBS) containing 10-5 M of TO and incubating this mixture for 15 min in the dark at room temperature. The stained cells may be analyzed fresh or after fixation with 1% paraformaldehyde/PBS or 0.25% may be fixed first and then stained. There is excellent correspondence between the number of fluorescent-labeled parasitized red cells and Giemsa-stained cells.