Request for BD FACSAria Fusion Cell Sorter ShEEP Application

该提案针对高容量流式细胞仪分选仪BD FACSAria融合细胞分选仪， 允许VA安阿伯卫生部的6名主要用户（优异奖资助的PI）和7名次要用户 护理系统（VAAAHS）进行多参数流式细胞术细胞分选， 纯化的细胞亚群用于随后的分析、实验操作和研究。我们 研究重点是优先影响人类健康的疾病的细胞和分子机制。 退伍军人：慢性阻塞性肺病、神经退行性疾病和中枢神经系统创伤、实体癌和白血病， 与部署有关的肺部疾病和一组感染性并发症， 风险更高。涉及特定的和经常罕见的细胞亚群及其独特的 越来越多的人发现，应对措施会影响这些条件，因此需要加以研究。新 实验方法，如单细胞RNA测序，细胞载体递送， 免疫方法和过继转移都依赖于高纯度细胞分选， 必须快速、精确和无菌地进行，以获得特定的细胞群， 高生存能力。这些需求在VAAAHS没有得到充分满足。要求的FACS 我们的VA站目前还没有一台分选机，它将允许收集稀有细胞群 基于它们表达的一组独特的多个特异性标记， 不可能或者很难得到。高容量分拣机将允许及时分拣 时尚（以维持细胞活力，无菌性和未改变的激活状态），同时工作 多个样本。我们多元化的用户群体将能够1）有效地收集定量数据 来自小而珍贵的样品的数据; 2）在进一步的测定中使用高度特异性的细胞亚群 如RNA转录组分析、单细胞测序、蛋白质组学、基因阵列分析等; 3） 产生用于过继转移实验和临床前治疗测试的细胞; 4）分离和 研究从临床样本和植入设备中纯化的细胞亚群，以发现新的 生物标志物、治疗靶点，以及推进治疗植入物的设计。引入 这些新的能力将提高目前研究的质量，提高效率， 吞吐量它也将提高我们的教师未来的赠款申请，并创造新的学习 为VAAAHS现场的受训人员提供机会，最终使退伍军人的健康受益。

This proposal is for a high capacity flow cytometry sorter BD FACSAria Fusion Cell Sorter to allow 6 Major Users (Merit Award funded PIs) and 7 Minor Users at the VA Ann Arbor Health Care System (VAAAHS) to conduct Multi-parameter flow cytometry cell sorting to isolate highly purified cell subsets for subsequent analysis, experimental manipulations, and studies. Our research focuses on cellular and molecular mechanisms of diseases that preferentially affect veterans: COPD, neurodegenerative disorders and CNS trauma, solid cancers and leukemias, the deployment-related lung disease and group of infectious complications for which veterans have higher risk. Involvement of specific and frequently rare cell subsets and their unique responses are increasingly found to influence these conditions and need to be studied. New experimental approaches, such as single-cell RNA sequencing, cellular vector deliveries for immunotherapeutic approaches, and adoptive transfers, all rely on high-purity cell sorting that must be performed rapidly, precisely, and aseptically to acquire specific cell populations with high viability. These needs are not being adequately met at the VAAAHS. Requested FACS sorter, presently unavailable at our VA station, will allow for collection of rare cell populations based on their expression of a unique set of multiple specific markers that would otherwise be impossible or very difficult to obtain. The high capacity sorter will allow for sorting in a timely fashion (to maintain cell viability, sterility and unaltered activation state) while working on multiple samples. Our diverse group of users will be able to 1) effectively collect quantitative data from small and precious samples; 2) use of highly specific subsets of cells in further assays like RNA transcriptome analysis, single-cell sequencing, proteomics, gene array analysis, etc; 3) generate cells for adoptive transfer experiments and preclinical therapy tests; 4) isolate and study purified cell subsets from clinical samples and implanted devices to discover new biomarkers, therapeutic targets, and to advance design of therapeutic implants. Introducing these new capabilities will enhance quality of current studies and increase efficiency and throughput. It will also enhance future grant applications of our faculty and create new learning opportunities for trainees at the VAAAHS site to ultimately benefit Veterans’ health.