Bigfoot Multispectral High Speed Fluorescence Activated Cell Sorter

Bigfoot 多光谱高速荧光激活细胞分选仪

• 批准号: 10414835
• 负责人: Steven A Porcelli
• 金额: $ 60万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-01 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10414835
• 关键词: Algorithms; Area; Attention; Basic Science; Cancer Center; Cells; Core Facility; Detection; Development; Dyes; Ensure; Equipment; Faculty; Financial compensation; Flow Cytometry; Fluorescence; Funding; Institution; Laboratories; Medicine; Minor; Monitor; Optics; Output; Population; Research Personnel; Research Project Grants; Sampling; Science; Sorting - Cell Movement; Speed; Supervision; Technology; Time; Translational Research; Tube; United States National Institutes of Health; base; cohort; college; design; detector; experience; flexibility; fluorescence activated cell sorter device; fluorophore; instrument; member; new technology; operation; optical spectra; senior faculty

We are requesting funds to assist us in purchasing a ThermoFisher Scientific Bigfoot spectral high speed cell sorter to expand and upgrade the equipment of the main Flow Cytometry Core Facility at the Albert Einstein College of Medicine. The Bigfoot is based on a remarkable new design that makes use of a principle known as full spectrum fluorescence analysis. This technology uses the full spectrum of emissions over a fixed detector array and an unmixing algorithm to greatly increase the number of fluorophores detected per cell, while eliminating most compensation issues that are major limitations of conventional flow cytometers. The optimized but flexible optical configuration makes the instrument robust and relatively easy to use, and permits detection of any fluorescence emission in the 400-900 nm range without requiring major adjustments of optical filters. The Bigfoot sorter’s optical design and unmixing algorithm enable the use of a wide array of dyes, including those with highly overlapping spectra—a feat not possible using conventional compensation-based cytometers and sorters. The problem of correct compensation of dyes with overlapping emission spectra, a major challenge for advanced multiparameter analysis, is mostly eliminated by the full spectrum approach of the Bigfoot. In addition to the unique spectral detection and processing, the Bigfoot provides extremely fast sort speeds for high yields and purity of specific cell populations. The instrument supports a range of sample outputs, including 6-way sorting into tubes and automated 8-way sorting into various format microtiter plates. A large cohort of NIH-funded investigators at Einstein working in diverse areas of biomedical science will benefit from access to this instrument, including the 24 major and 14 minor users specifically identified in this application. We plan to house and operate the Bigfoot sorter in our well established main Flow Cytometry Core Facility, where there will be ample expertise, oversight and attention to biosafety issues to ensure that the instrument is used correctly and to its full capacity. The Core Facility has an experienced senior faculty member (Dr. Steven Porcelli) as its supervisor, and employs two full time faculty level associates and two technicians to direct operations and provide technical supervision. There is also a range of internal consultants with expertise in areas of major relevance to the development of applications that will use the Bigfoot cell sorter. The Core Facility has a well-developed and carefully monitored financial plan, and receives support from the institution and from our NCI-funded Cancer Center to ensure its efficient and stable operation. By incorporating the Bigfoot spectral high speed cell sorter into this core laboratory, a large body of investigators pursuing diverse NIH-funded basic and translational research projects will have convenient and affordable access to this important new technology.

我们正在申请资金来帮助我们购买 ThermoFisher Scientific Bigfoot 光谱高速 细胞分选机用于扩展和升级阿尔伯特主要流式细胞术核心设施的设备 爱因斯坦医学院。 Bigfoot 基于一项非凡的新设计，该设计利用了 原理称为全光谱荧光分析。该技术使用全谱发射 通过固定检测器阵列和分离算法大大增加荧光团的数量 检测每个细胞，同时消除大多数补偿问题，这是传统流程的主要限制 细胞仪。优化但灵活的光学配置使仪器坚固且相对简单 使用，并允许检测 400-900 nm 范围内的任何荧光发射，无需大量 光学滤镜的调整。 Bigfoot 分拣机的光学设计和分解算法使得能够使用 广泛的染料，包括那些具有高度重叠光谱的染料——这是使用传统技术不可能实现的壮举 基于补偿的细胞仪和分选仪。染料的正确补偿问题 重叠发射光谱是先进多参数分析的主要挑战，现已基本消除 通过大脚怪的全方位方法。除了独特的光谱检测和处理之外， Bigfoot 提供极快的分选速度，可实现特定细胞群的高产量和高纯度。这 仪器支持一系列样品输出，包括 6 路分选管和自动 8 路分选 分类成各种规格的微量滴定板。爱因斯坦的一大批由 NIH 资助的研究人员在 生物医学科学的各个领域都将受益于该仪器，包括 24 个主要和 本申请中特别确定了 14 名未成年用户。我们计划在以下地点安置和运营 Bigfoot 分拣机 我们完善的主要流式细胞术核心设施，将拥有充足的专业知识、监督和 注意生物安全问题，以确保仪器正确使用并充分发挥其作用。核心 该设施拥有一位经验丰富的高级教员（Steven Porcelli 博士）作为其主管，并雇用了两名 全职教职人员和两名技术人员指导操作并提供技术监督。 还有一系列在与开发主要相关领域具有专业知识的内部顾问 将使用 Bigfoot 细胞分选仪的应用程序。核心设施拥有完善且精心设计的 监控财务计划，并获得该机构和我们 NCI 资助的癌症中心的支持 确保其高效稳定运行。通过将 Bigfoot 光谱高速细胞分选仪纳入 这个核心实验室拥有大量研究人员，致力于 NIH 资助的各种基础和转化研究 研究项目将能够方便且经济地获得这一重要的新技术。