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

项目摘要 Fluoropolitan，LLC正在开发Fluoropolitan智能面板构建器，这是一种用于 自动计算最佳荧光染料细胞计数板。流式细胞术是一种普遍存在的技术， 细胞和分子生物学由于最近仪器性能的改进而日益复杂。 研究人员通常会设计出一个包含20多种颜色的细胞仪面板， 实验为了成功设计多参数细胞仪面板，研究人员必须将设计 优化荧光染料激发/发射溢出、优化荧光染料亮度的最佳实践 他们必须从广泛而多样的抗体市场中选择抗体。两 专家和非专家将受益于一种自动计算最佳面板的方法， 它们的仪器配置和实验生物标志物。Fluoroelasticity以前开发了一种 手动面板生成器，包含广泛的多供应商数据库，包括所有商业 现有的仪器、所有荧光染料和超过300万种抗体，以简化面板设计的各个方面。 在本提案中，Fluoropilot将利用这些广泛的数据库来构建智能面板构建器。的 智能面板生成器将通过考虑用户特定的 实验配置，采用用户和实验特定的门控策略，并定量 最小化荧光溢出。预计这将大大减少开发复杂细胞仪的时间 面板和显着减少或潜在地消除设计中的错误。预计这也将减少 对于参与流式细胞术的非专家研究人员来说，这是一个进入门槛。第一阶段提案的目标 是在专家细胞测量师的支持下重建数据库并开发算法， 比较荧光染料（目标1），通过在Intellipanel上调查一组细胞计数专家来证明疗效 生成面板（目标2），并开发直观的Web应用程序，以支持简化的用户体验 (Aim 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).