High-parametric flow cytometry cell sorter for human immunology studies.

项目摘要 流式细胞术是分析组织中免疫细胞群的最常用和最关键的工具， 能够实现约18-20个参数的群体分离并维持细胞活力。通过传递细胞 通过施加激光激发的流，可以将单个细胞群分离成 96孔板作为单细胞，使得能够克隆和其他应用，或者可以作为群体在4-6个细胞中富集。 不同的人群同时这是唯一的细胞分离技术普遍适用于多个 组织，包括从困难部位（如外周组织、坏死肿瘤）分离罕见群体，以及 机关 目前，威尔康奈尔医学院（WCMC）还不能随时获得这项尖端技术， 医学中心研究社区。我们目前的流式细胞仪核心是在最大容量使用的基本 科学应用程序，并提前几周到几个月预订，禁止与临床应用程序的需求保持一致。 研究人员和基础和转化研究直接对患者样本。 基于对WCMC科学家的前瞻性调查，我们还确定了超过12个NIH资助的主要用户， 作为来自多个部门的大量小用户， 他们的工作。拟议的应用是多种多样的，利用了一些国家的各种科学优势， WCMC实验室。一些例子包括阿里·梅尔尼克博士对人类B细胞生殖中心的理解， 博士Sallie Permar对儿科感染和人类疫苗反应的理解以及 保护性免疫，Laura Santambrogio博士正在进行的关于人类MHC肽组的研究， 免疫抗原呈递，以及布拉德琼斯博士对HIV发病机制和T细胞免疫的研究 应答 世界养护监测中心的领导层强烈支持购置这一仪器，并将为该仪器提供重要的 在S10申请中要求的额外资金。此外，还分配了专用空间， Belfer研究大楼和人类免疫监测中心内，仪器将位于 并管理。总的来说，我们假设在细胞上建立流式细胞术细胞分选仪， WCMC校园将在许多项目和学科中推动新的科学见解。

PROJECT SUMMARY Flow cytometry is the most commonly used and critical tool for analyzing immune cell populations in tissues, enabling approximately 18-20 parameter separation of populations and maintaining cell viability. By passing cells through a stream in which laser excitation is applied, individual populations of cells can be separated into either 96 well plates as single cells, enabling cloning and other applications, or can be enriched as populations in 4-6 separate populations simultaneously. This is the only cell separation technology universally adaptable to multiple tissues including isolation of rare populations from difficult sites such as peripheral tissues, necrotic tumors, and organs. Currently, this cutting-edge technology is not readily available to the Weill Cornell Medical College (WCMC) and medical center research community. Our current flow cytometry core is at maximum capacity usage for basic science applications and is booked weeks to months in advance, prohibiting alignment to the needs of clinical research investigators and basic and translational research directly on patient samples. Based on a prospective survey of WCMC scientists, we have identified over 12 NIH-funded major users, as well as a large number of minor users from multiple departments who have requested this instrumentation to advance their work. The proposed applications are diverse, leveraging the various scientific strengths of a number of WCMC labs. Some examples include Dr. Ari Melnick's work on understanding human B cell germinal centers, Dr. Sallie Permar’s understanding of pediatric infections and human vaccine responses and correlates of protective immunity, Dr. Laura Santambrogio’s ongoing studies on the human MHC peptidome to understand immune antigen presentation, and Dr. Brad Jones’ investigations into HIV pathogenesis and T cell immune responses. The leadership of WCMC is strongly supportive of the acquisition of this instrument, and will provide significant additional funds beyond those requested in the S10 application. In addition, dedicated space has been allocated within the Belfer Research Building and Human Immune Monitoring Core, where the instrument will be located and managed. Overall, we hypothesize that the establishment of a Flow Cytometry cell sorter instrument on the WCMC campus will be pivotal in driving new scientific insights across a number of projects and disciplines.