Bigfoot Multispectral High Speed Fluorescence Activated Cell Sorter

我们正在申请资金，以帮助我们购买一台ThermoFisher Science BigFoot光谱高速 细胞分选机将扩建和升级阿尔伯特主流细胞测量核心设施的设备 爱因斯坦医学院。大脚怪是基于一种引人注目的新设计，它利用了 原理称为全光谱荧光分析。这项技术使用了排放的全光谱 通过固定的探测器阵列和分解算法来大大增加荧光团的数量 同时消除了作为传统流程的主要限制的大多数补偿问题 细胞仪。优化但灵活的光学配置使仪器坚固耐用，相对容易 使用，并允许检测400-900 nm范围内的任何荧光发射，而不需要 滤光片的调整。BigFoot分类器的光学设计和拆分算法使其能够使用 广泛的染料系列，包括那些光谱高度重叠的染料--这是使用传统方法无法实现的壮举 基于补偿的细胞仪和分选机。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，基本消除了 通过大脚怪的全光谱方法。除了独特的光谱检测和处理外， 大脚怪提供了极快的分拣速度，以获得高产量和特定细胞群的纯度。这个 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 分选成各种格式的微滴定板。一大批由美国国立卫生研究院资助的爱因斯坦研究人员在 生物医学科学的不同领域将受益于获得这一仪器，包括24个主要和 此应用程序中明确标识的14个次要用户。我们计划将大脚怪分拣机安置在这里并进行操作 我们建立了完善的主流细胞计数核心设施，在那里将有充足的专业知识、监督和 注意生物安全问题，以确保正确使用该仪器并充分发挥其能力。《核心》 设施有一位经验丰富的资深教员(史蒂文·波塞利博士)作为其主管，并雇用了两名 全职教员级别的助理和两名技术人员指导操作并提供技术监督。 还有一系列内部顾问，他们在与发展密切相关的领域具有专门知识。 将使用大脚细胞分类器的应用程序。核心设施有一个开发得很好和仔细的 监控财务计划，并得到该机构和我们NCI资助的癌症中心的支持 以确保其高效稳定运行。通过将BigFoot光谱高速细胞分选器整合到 这个核心实验室，一个由大量研究人员组成的团队，致力于研究由NIH资助的各种基础和翻译 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。

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.