Spectral Configured Bigfoot Sorter for Salk Institute Flow Cytometry Core

用于索尔克研究所流式细胞术核心的光谱配置 Bigfoot 分选机

• 批准号: 10632950
• 负责人: Carolyn Pey-Min O'Connor
• 金额: $ 72.45万
• 依托单位: SALK INSTITUTE FOR BIOLOGICAL STUDIES
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10632950
• 关键词: 10 year old; Acceleration; Address; Aerosols; Aging; Air; Antigens; Applications Grants; Basic Science; Biological; Biological Sciences; Biology; Cancer Biology; Cells; Containment; Deposition; Detection; Failure; Flow Cytometry; Fluorescence; Immunology; Lasers; Life; Mechanics; Modernization; Neurosciences; Pathogenesis; Phenotype; Plants; Regenerative Medicine; Research; Research Personnel; Research Support; Resource Sharing; Sampling; Sorting; Speed; Stains; System; Technology; Time; cell type; cost; design; detector; experience; fluorophore; instrument; instrumentation; interest; microbial; migration; nervous system disorder; new technology; programs

Project Summary The Salk Institute for Biological Studies is an independent non-profit institute for basic science research located in La Jolla, CA. Salk research programs include cancer biology, immunology and microbial pathogenesis, aging and regenerative medicine, neuroscience and neurological disorders, and plant biology. Shared resources and technology cores provide critical support for this research. The Flow Cytometry Core provides Institute researchers access to flow cytometry instrumentation. To accommodate increasing demands for sorting, multi- year strategic acquisitions have been implemented, effectively doubling the Core’s sorting instrumentation since 2017 (instruments were added in 2017, 2018 and 2021). At present, all four sorters in the Core are highly utilized. Three sorters are BD Biosciences Influx instruments, which feature jet-in-air fluidic design and high-speed, multi- well plate deposition mechanics. These Influxes have been versatile and dependable workhorses for many Salk projects over many years. Unfortunately, the Influx system has reached the end of its useful life, with the Core’s units being on average ~10 years old. Costly experimental failures, reduced access to technical support, and a lack of solutions for replacement parts have collectively escalated the urgency to migrate to newer technology, in particular as sorting demands at Salk are projected to continue to increase. Additionally, two Influxes do not meet current standards for aerosol containment. The purpose of this grant application is to address an urgent need: to upgrade the Core’s sorting capabilities by acquiring a cutting-edge modern cell sorter. The Core proposes to acquire a Thermo Fisher Spectral Bigfoot (5-laser, 48-fluorescence detectors). The new instrument will supplement current sorting capacity and be invaluable in times of unexpected delays due to technical failure. The Bigfoot is a best-in-class “jet-in-air” type cell sorter and is the only viable option to replace the obsolete jet-in-air Influx systems. No extra accessories are needed: Bigfoot has an integrated Class II biocontainment enclosure, aerosol containment features, and a high-speed multi-well deposition system for plate sorting. The proposed instrument will use a “spectral” configuration that has the 5 common laser lines that are used in Salk phenotyping panels. This 5-laser spectral configuration will allow investigators to make use of previously optimized staining panels and will add powerful “spectral detection” capabilities currently unavailable at Salk. A major advantage of spectral detection capability is that it provides access to fluorophore combinations previously not possible. This in turn allows additional antigens of interest to be easily added to existing panels. Taken together, there are significant cost and time benefits for leveraging spectral detection: more information can be collected from limited, precious samples, thereby accelerating scientific discovery. The Salk Institute has a strong track record of demonstrated commitment to the implementation, support, and long-term management of high-end instrumentation. With the experience and expertise provided by the Salk Flow Cytometry Core, this new instrument will be immediately utilized at a high level, thereby facilitating scientific discovery at the Institute.

项目摘要 SALK生物学研究所是一个独立的非营利基础科学研究所 在加利福尼亚州拉霍亚SALK研究计划包括癌症生物学，免疫学和微生物发病机理，衰老 以及再生医学，神经科学和神经系统疾病以及植物生物学。共享资源和 技术核心为这项研究提供了重要的支持。流式细胞仪核心提供了研究所 研究人员可以使用流式细胞仪仪器。为了适应对分类的不断增长的需求 已经实施了一年的战略收购，自从核心的分类工具有效地翻了一番 2017年（在2017年，2018年和2021年添加了仪器）。目前，核心中的所有四个分类器都高度利用。 三个分类器是BD Biosciences涌入仪器，它们具有喷射液体设计和高速，多速度 板板沉积力学。这些涌入的多功能性和依赖于许多salk 多年的项目。不幸的是，涌入系统已经结束了其用途的终点 单位平均约10年。昂贵的实验失败，减少获得技术支持的机会，并且 缺乏替换零件的解决方案已统一升级了迁移到新技术的紧迫性， 特别是因为Salk的分类需求预计将继续增加。此外，两次涌入没有 符合当前的气溶胶遏制标准。本赠款申请的目的是解决 迫切需要：通过辅助尖端的现代细胞分系程序来升级核心的分类功能。 核心提议获得了热渔民光谱Bigfoot（5射线器，48-荧光探测器）。 新仪器将补充当前分类能力，并在意外延迟时无价 技术故障。 Bigfoot是一流的“空中喷射”类型分隔器，是唯一更换的可行选择 过时的喷气发入系统。不需要额外的配件：Bigfoot拥有II类集成 生物膜片机箱，气溶胶遏制功能和板的高速多孔沉积系统 排序。所提出的仪器将使用具有5个常见激光线的“光谱”配置 用于SALK表型面板。这种5射线光谱配置将允许调查人员使用 先前优化的染色面板，并将添加目前无法使用的功能强大的“光谱检测”功能 在萨克。光谱检测能力的主要优点是它提供了荧光团组合的访问 以前是不可能的。反过来，这允许将其他感兴趣的抗原轻松添加到现有面板中。 综上 可以从有限的珍贵样品中收集，从而加速科学发现。 Salk Institute有 对实施，支持和长期管理的表现出色的有力记录 高端仪器。凭借Salk流式细胞仪核提供的经验和专业知识， 新仪器将立即在高水平上使用，从而支持该研究所的科学发现。