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 索尔克生物研究所是一个独立的非营利性基础科学研究机构，位于 在加利福尼亚州拉荷亚。SALK研究项目包括癌症生物学、免疫学和微生物发病机制、衰老 以及再生医学，神经科学和神经紊乱，以及植物生物学。共享资源和 技术核心为这项研究提供了关键支持。流式细胞仪核心提供研究所 研究人员可获得流式细胞仪仪器。为了适应日益增长的分类需求，多个 年实施了战略收购，有效地将Core的分拣工具翻了一番 2017年(2017年、2018年和2021年增加了工具)。目前，核心中的所有四个分拣机都得到了高度利用。 三台分选机均为BD Biosciences Inflation仪器，采用空气喷射流态化设计，具有高速、多通道、多通道的特点。 好的，平板沉积力学。这些涌入对许多Salk来说是多才多艺和可靠的主力 多年来的项目。不幸的是，流入系统已经到了其使用寿命的尽头，核心的 单位平均使用年限约为10年。代价高昂的试验性故障、获得技术支持的机会减少，以及 缺乏更换部件的解决方案共同加剧了向更新技术迁移的紧迫性， 特别是由于索尔克的分拣需求预计将继续增加。此外，两次涌入不会 符合当前的气溶胶控制标准。这项赠款申请的目的是解决一个 迫切需要：通过收购尖端的现代细胞分选器来升级Core的分选能力。 核心计划购买一台Thermo Fisher Spectral BigFoot(5台激光、48台荧光探测器)。这个 新仪器将补充目前的分拣能力，在因以下原因而出现意外延误时具有无价价值 技术故障。大脚是一种最好的“空中喷射”式细胞分选机，是唯一可行的选择，以取代 过时的空气喷射系统。不需要额外的配件：大脚野人有一个集成的II级 生物围护结构、气溶胶围护结构和用于平板的高速多孔沉积系统 分类。建议的仪器将使用一种“光谱”配置，它有5条常见的激光线路， 用于Salk表型面板。这种5激光光谱配置将允许调查人员利用 以前优化过的染色面板，并将增加目前无法提供的强大的“光谱检测”功能 在索尔克。光谱检测能力的一个主要优势是它提供了访问荧光团组合的途径 以前是不可能的。这反过来又允许将更多感兴趣的抗原轻松地添加到现有的面板中。 总而言之，利用光谱检测具有显著的成本和时间效益：更多信息 可以从有限的、珍贵的样本中收集，从而加速科学发现。索尔克研究所已经 表现出致力于实施、支持和长期管理的良好记录 高端仪器的市场。凭借Salk Flow细胞测量核心提供的经验和专业知识，这 新仪器将立即在高水平上使用，从而促进研究所的科学发现。