High-throughput single-cell analysis in microfluidically-generated droplets

该项目是在本财政年度结束时在VRC免疫学实验室新成立的一个部门启动的，目的是利用微流体技术在另一个项目中回答一个特定的生物学问题。该项目的基本原理是，下一代测序的新兴技术允许对基因组、转录组和其他复杂的核酸群体进行全面测定，但它们本身不允许对复杂群体中单个细胞的信号进行分辨率。在需要细胞间异质性信息的多种情况下，这是一个重要的限制。这包括对大量群体中罕见病毒感染细胞的分析，以及对高度多样化和复杂群体的全球理解。

This project was initiated at the end of the fiscal year in a newly-created section of the Immunology Laboratory, VRC to leverage work using microfluidics undertaken to answer a specific biological question in a separate project. The overarching rationale for this project is that emerging technologies for next-generation sequencing allow the comprehensive determination of genomes, transcriptomes, and other complex populations of nucleic acid, but on their own do not permit resolution of signal from individual cells within complex populations. This is a significant limitation in multiple settings where information about intercellular heterogeneity is needed. These include the analysis of rare virus-infected cells in large populations as well as the global understanding of highly diverse and complex populations. To address this need, this project seeks to employ microfluidics to generate stable, uniform emulsions in which single cells are encapsulated at limiting dilution. Thus, tens of millions of single-cell reaction chambers can be created. Various adaptations of next-generation sequencing library preparation technology are then employed to generate material for sequencing that capture information about intercellular heterogeneity within cell populations studied.