We developed a flow-cytometry-based method to separate and collect cocultured male and female Plasmodium falciparum gametocytes responsible for malaria transmission. The purity of the collected cells was estimated at >97% using flow cytometry, and sorted cells were observed by Giemsa-stained thin-smear and live-cell fluorescence microscopy. The expression of validated sex-specific markers corroborated the sorting strategy. Collected male and female gametocytes were used to confirm three novel sex-specific markers by quantitative real-time PCR that were more enriched in sorted male and female gametocyte populations than existing sex-specific markers. We also applied the method as a proof-of-principle drug screen that allows the identification of drugs that kill gametocytes in a sex-specific manner. Since the developed method allowed for the separation of male and female parasites from the same culture, we observed for the first time a difference in development time between the sexes: females developed faster than males. Hence, the ability to separate male and female gametocytes opens the door to a new field of sex-specific P. falciparum gametocyte biology to further our understanding of malaria transmission.

中文翻译：

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分离疟原虫恶性疟原虫的雄性和雌性配子体的新方法，使生物和药物发现成为可能。

我们开发了一种基于流式细胞术的方法来分离和收集共培养的雄性和雌性恶性疟原虫负责疟疾传播的配子体。使用流式细胞术估计收集到的细胞纯度 > 97%，并通过吉姆萨染色薄涂片和活细胞荧光显微镜观察分选的细胞。经验证的性别特异性标记的表达证实了排序策略。收集到的雄性和雌性配子体用于通过实时定量 PCR 确认三个新的性别特异性标记物，这些标记物在分选的雄性和雌性配子体种群中比现有的性别特异性标记物更丰富。我们还应用该方法作为原理验证药物筛选，允许鉴定以性别特异性方式杀死配子体的药物。由于开发的方法允许从同一文化中分离雄性和雌性寄生虫，我们第一次观察到两性之间发育时间的差异：女性比男性发育得更快。因此，分离男性和女性配子体的能力打开了通向特定性别的新领域的大门。恶性疟原虫配子体生物学以进一步了解疟疾传播。