Makler M T, Lee L G, Recktenwald D
Laboratory Service, Veterans Administration Medical Center, Portland, Oregon 97201.
Cytometry. 1987 Nov;8(6):568-70. doi: 10.1002/cyto.990080606.
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% glutaraldehyde/PBS. Alternatively the cells may be fixed first and then stained. There is excellent correspondence between the number of fluorescent-labeled parasitized red cells and Giemsa-stained cells.
本文介绍了一种用于测定体外培养的恶性疟原虫的快速灵敏方法。该技术采用配备15毫瓦氩激光（发射488纳米波长的光）的荧光流式细胞仪和一种可透过细胞膜的荧光染料噻唑橙（TO），它能对RNA进行染色。将寄生有疟原虫的红细胞悬浮于1毫升含10^(-5) M TO的磷酸盐缓冲盐水（PBS）中进行染色，并将该混合物在室温黑暗条件下孵育15分钟。染色后的细胞可直接分析，也可用1%多聚甲醛/PBS或0.25%戊二醛/PBS固定后再分析。或者先固定细胞，然后再进行染色。荧光标记的寄生有疟原虫的红细胞数量与吉姆萨染色的细胞数量之间具有良好的对应关系。
