Improved flow cytometry panel design with a novel automated panel design system

项目总结 FluoroFinder，LLC正在开发FluoroFinder Intellipanel Builder，这是一种创新的软件系统，用于 最佳荧光细胞仪板的自动计算。流式细胞术是一种普遍存在的技术 由于最近仪器能力的提高，细胞和分子生物学变得越来越复杂。 对于研究人员来说，设计一块包含20多种颜色的细胞仪面板是很常见的 做实验。为了成功地设计多参数细胞仪仪表板，研究人员必须将设计 优化荧光激发/发射溢出、优化荧光亮度的最佳实践 通过抗原密度，他们必须从广泛和多样化的抗体市场中选择抗体。两者都有 专家和非专家将受益于一种自动计算最优面板的方法 关于它们的仪器配置和实验生物标记物。FluoroFinder之前开发了一种 手动面板生成器，集成了一个广泛的多供应商数据库，其中包括所有商业 可用的仪器、所有荧光染料和300多万种抗体，以简化面板设计的方方面面。 在这份提案中，FluoroFinder将利用这些广泛的数据库来构建IntelliPanel生成器。这个 IntelliPanel Builder将为用户的实验生成一个最佳面板，方法是考虑到用户特定的 实验配置，采用用户和实验特定的门控策略，并定量 最大限度地减少荧光溢出。预计这将大大减少开发复杂细胞分析的时间 并显著减少或潜在地消除设计中的错误。预计这也将减少 参与流式细胞术的非专家研究人员的准入门槛。此第一阶段提案的目标 是在专业细胞学家的支持下重组数据库和开发算法，以量化 比较荧光染料(目标1)，通过调查Intellipanel上的一组细胞学专家来演示有效性 生成的面板(Aim 2)，并开发直观的Web应用程序以支持简化的用户体验 (目标3)。

PROJECT SUMMARY FluoroFinder, LLC is developing the FluoroFinder Intellipanel builder, an innovative software system for the automatic calculation of an optimal fluorochrome cytometry panel. Flow cytometry is a ubiquitous technique in cell and molecular biology that is increasingly complex due to recent improvements in instrumental capabilities. It is common for researchers to design cytometry panels that incorporate more than 20-colors in a single experiment. To successfully design a multi-parameter cytometry panel, researchers must incorporate design best practices to optimize fluorochrome excitation/emission spillover, optimize for fluorochrome brightness through antigen density, and they must select antibodies from an extensive and diverse antibody market. Both experts and non-experts would benefit from an approach that automatically calculates an optimal panel based on their instrumental configuration and experimental biomarkers. FluoroFinder has previously developed a manual panel builder that incorporates an extensive multi-supplier database that includes all commercially available instruments, all fluorochromes, and over 3 million antibodies to streamline all aspects of panel design. In this proposal, FluoroFinder will leverage these extensive databases to build the Intellipanel builder. The Intellipanel builder will generate an optimal panel for a user’s experiment by accounting for user specific experimental configurations, employing user and experimental specific gating strategies, and quantitatively minimizing fluorescent spillover. This is expected to significantly reduce the time to develop complex cytometry panels and significantly reduce or potentially eliminate errors in design. This is also expected to reduce the barrier to entry for non-expert researchers that are involved in flow cytometry. The goals of this Phase I proposal are to restructure databases and develop algorithms with the support of expert cytometrists to quantitatively compare fluorochromes (Aim 1), demonstrate efficacy by surveying a panel of cytometry experts on Intellipanel generated panels (Aim 2), and develop an intuitive web application to support a streamlined user experience (Aim 3).