Influenza A virus (IAV) is an RNA virus with high genetic diversity which necessitates the development of new vaccines targeting emerging mutations each year. As IAV exists in genetically heterogeneous populations, current studies focus on understanding population dynamics at the single cell level. These studies include novel methodology that can be used for probing populations at the single cell level, such as single cell sequencing and microfluidics. Here, we introduce a drop-based microfluidics method to study IAV infection at a single cell level by isolating infected host cells in microscale drops. Single human alveolar basal epithelial (A549), Madin-Darby Canine Kidney cells (MDCK) and MDCK + human siat7e gene (Siat7e) cells infected with the pandemic A/California/07/2009 (H1N1) strain were encapsulated within 50 μm radii drops and incubated at 37°C. We demonstrate that drops remain stable over 24 hours, that 75% of cells remain viable, and that IAV virus can propagate within the drops. Drop-based microfluidics therefore enables single cell analysis of viral populations produced from individually infected cells.

中文翻译：

---

使用基于液滴的微流体技术感染甲型流感病毒的单细胞

甲型流感病毒 (IAV) 是一种具有高度遗传多样性的 RNA 病毒，因此每年都需要开发针对新出现的突变的新疫苗。由于 IAV 存在于遗传异质群体中，目前的研究侧重于了解单细胞水平的群体动态。这些研究包括可用于在单细胞水平上探测种群的新方法，例如单细胞测序和微流体。在这里，我们介绍了一种基于液滴的微流体方法，通过在微尺度液滴中分离受感染的宿主细胞，在单细胞水平上研究 IAV 感染。单个人肺泡基底上皮 (A549)、Madin-Darby 犬肾细胞 (MDCK) 和 MDCK + 人siat7e感染了大流行 A/California/07/2009 (H1N1) 毒株的基因 (Siat7e) 细胞被封装在 50 μm 半径滴内，并在 37°C 下孵育。我们证明液滴在 24 小时内保持稳定，75% 的细胞保持活力，并且 IAV 病毒可以在液滴内传播。因此，基于液滴的微流体技术可以对由单个感染细胞产生的病毒群进行单细胞分析。