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，以帮助我们购买赛默飞世尔科学大脚光谱高速 细胞分选机，以扩大和升级的主要流式细胞术核心设施的设备在阿尔伯特 爱因斯坦医学院。大脚怪是基于一个显着的新设计，利用了一个 原理称为全光谱荧光分析。这项技术利用了全光谱的辐射 通过固定的检测器阵列和解混算法， 每个细胞检测，同时消除了大多数补偿问题，这是传统流量的主要限制 细胞计数器。优化但灵活的光学配置使仪器坚固耐用且相对简单 并允许检测400-900 nm范围内的任何荧光发射，而不需要主要的 光学滤光片的调整。大脚分类器的光学设计和解混算法使得能够使用 广泛的染料，包括那些高度重叠的光谱-一个壮举不可能使用传统的 基于补偿的细胞仪和分选仪。染料的正确补偿问题 重叠的发射光谱是先进的多参数分析的主要挑战， 大脚怪的全方位攻击除了独特的光谱检测和处理之外， Bigfoot提供了极快的分选速度，以获得高产量和高纯度的特定细胞群。的 仪器支持一系列样品输出，包括6路分选到试管中和自动8路分选到试管中。 分选到各种规格的微量滴定板中。一大批由NIH资助的研究人员在爱因斯坦工作， 生物医学科学的各个领域将受益于这一工具，包括24个主要领域， 14个未成年用户在本申请中明确指出。我们计划将大脚分类机安置在 我们完善的主要流式细胞术核心设施，在那里将有足够的专业知识，监督和 注意生物安全问题，以确保正确使用仪器并充分发挥其能力。核心 设施有一个经验丰富的高级教员（博士史蒂芬Porcelli）作为其主管，并雇用两名 全职教员水平的助理和两名技术人员，以指导业务和提供技术监督。 还有一系列内部顾问，他们在与发展相关的主要领域拥有专业知识。 将使用大脚细胞分类器的应用程序。核心设施有一个良好的发展和仔细 受监控的财务计划，并获得该机构和我们的NCI资助的癌症中心的支持 以确保其高效、稳定的运行。通过将大脚光谱高速细胞分选机整合到 这个核心实验室，一个庞大的研究机构，追求不同的NIH资助的基础和翻译 研究项目将可以方便和负担得起地使用这项重要的新技术。