Magnetically directed single cell transcriptome analysis in HIV latency

 描述(申请人提供)：该项目利用生物医学和工程研究人员之间的新合作来开发新的免疫细胞分类和分离方法，其中单个细胞的精确轨迹在芯片上进行控制和分类，类似于计算机电路中电子的控制方式。这套工具允许使用全新的方法来研究单个或成对单细胞的表型、基因和形态，在更长的时间内具有现有技术所无法比拟的精确度，如流式细胞仪和流式细胞分选。这项研究的总体目标是开发工程平台并展示芯片实验室设备的生物学应用，该设备可以长时间分析数千个单细胞，暴露在多种刺激下，并能够提取单个、高价值的细胞用于进一步的免疫学分析(RT-PCR、克隆扩增等)。我们的目标是1)设计和制造一种针对生物应用而优化的芯片上实验室检测方法，2)针对生物应用优化该检测方法，以及3)使用这种新的检测方法来研究HIV潜伏期中断中的早期细胞事件。

 DESCRIPTION (provided by applicant): This project leverages new collaborations between biomedical and engineering investigators to develop new methods for the sorting and isolation of immune cells, where the precise trajectory of individual cells are controlled and sorted on a chip analogous to the way electrons are controlled inside computer circuits. This set of tools allows for fundamentally new methods to study phenotype, genotype, and the morphology of single or pairs of single cells, over extended periods of time with precision that is unparalleled by existing techniques, such as flow cytometry and flow cell sorting. The overall stated aims of this research are to develop the engineering platform and demonstrate a biological application for a lab-on-a-chip device that can analyze thousands of single cells over long durations, exposure to multiple stimuli, and enable the extraction of individual, high-value, cells for furthe immunological analyses (RT-PCR, clonal expansion, etc.). We aim to 1) design and fabricate a lab-on-chip assay optimized for biological applications, 2) optimize the assay for biological applications, and 3) use this novel assay to study early cellular events in the disruption of HIV latency.