Multimensional Robotic Flow Platform

使用流式细胞术的免疫表型分析与十年前的基因组学相似。具有6至18种颜色的高度多色测定已在学术实验室中得到证明，但通常没有以稳健的工业化方式定期部署用于临床研究。人类免疫系统监测，包括复杂的多参数流式细胞术的应用，将使更直接反映疾病发病机制和进展的深入表型分析成为可能。多色流式细胞术基于细胞数量、细胞类型以及与免疫扰动相关的细胞相关表面和细胞内分子的表达的差异，提供了对免疫功能的有力评估。开发一个全面的平台，引入：自动化细胞处理;在六分钟内运行96孔板的高速细胞询问;以及实施新开发的“FLAME”程序的多维数据分析工具，将允许精确和稳健地测量人类对病毒疫苗和感染的免疫反应。该核心的目的是为U19带来新技术，以实现高通量，同时在免疫或感染之前，期间和之后对免疫细胞状态进行稳健和精确的检查。将使用最先进的机器人平台，通过流式细胞术处理和分析从项目1和2中采集的大部分样本。多维机器人流动核心能够使用经验证的机器人方法来处理血液样本，从而使细胞制备的变化最小。将使用机器人平台制备处理后的样本，以使用多色流式细胞术检测不同细胞群的免疫状态标志物水平。机器人技术的应用，试剂的仔细质量控制，以及多维空间中流动数据的自动下游分析，对于U19建立人类免疫学分析研究小组网络的挑战至关重要。

Immunophenotyping using flow cytometry is in a position similar to that of genomics a decade ago. Highly polychromatic assays with 6 to 18 colors have been demonstrated in academic labs, but have generally not been regularly deployed for clinical studies in a robust, industrialized manner. Immune system monitoring in humans, including the application of sophisticated multi-parameter flow cytometry, would make possible indepth phenotyping that more directly reflects disease pathogenesis and progression. Polychromatic flow cytometry provides a powerful assessment of immune function based on differences in cell numbers, cell types and the expression of cell-associated surface and intracellular molecules related to immune perturbation. Development of a comprehensive platform that introduces: automation to cell processing; highspeed cell interrogation running a 96-well plate in under six minutes; and multidimensional data analysis tools implementing the newly developed "FLAME" program will allow the precise and robust measurement of human immune responses to viral vaccines and infections. The aim of this core is to bring new technologies to the U19 to achieve high throughput but also robust and precise examination of the immune cell status before, during and after immunization or infection. The bulk of all samples collected from Projects 1 and 2 will be processed and analyzed by Flow Cytometry using a state-of-the-art robotics platform. The multidimensional robotic flow core has the capability to use validated robotic methods to process blood samples, resulting in minimal variation in cell preparation. The processed samples will be prepared using the robotic platform to detect the level of immune status markers of different cell populations using polychromatic flow cytometry. The application of robotics, careful quality control of reagents, and automated downstream analysis of flow-data in multi-dimensional space is critical in the U19's challenge to establish this network of human immunology profiling research groups.