Multimensional Robotic Flow Platform

使用流式细胞术进行免疫表型鉴定的情况类似于十年前的基因组学。有6到18种颜色的高度多色分析已经在学术实验室中得到证实，但通常还没有以稳健的工业化方式定期用于临床研究。对人类免疫系统的监测，包括复杂的多参数流式细胞术的应用，将使深入的表型成为可能，更直接地反映疾病的发病机制和进展。多色流式细胞术基于细胞数量、细胞类型以及与免疫紊乱相关的细胞表面和细胞内分子表达的差异，提供了一种强大的免疫功能评估方法。开发一个全面的平台，引入：细胞处理的自动化；在6分钟内运行96孔板的高速细胞询问；以及实施新开发的“FLAME”计划的多维数据分析工具，将允许精确和强大地测量人类对病毒疫苗和感染的免疫反应。这一核心的目的是将新技术引入U19，以实现高吞吐量，并在免疫或感染之前、期间和之后对免疫细胞状态进行稳健和精确的检查。从项目1和2收集的大部分样本将通过使用最先进的机器人平台的流式细胞仪进行处理和分析。多维机器人流动核心具有使用经过验证的机器人方法处理血液样本的能力，从而使细胞准备过程中的差异最小。处理后的样本将使用机器人平台进行准备，以使用多色流式细胞术检测不同细胞群体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化下游分析，在U19‘S挑战中至关重要，以建立这个人类免疫图谱研究小组的网络。

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.