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.

项目概要 索尔克生物研究所是一家独立的非营利性基础科学研究机构，位于 位于加利福尼亚州拉霍亚。索尔克研究项目包括癌症生物学、免疫学和微生物发病机制、衰老 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 技术核心为本研究提供了关键支撑。流式细胞术核心提供研究所 研究人员可以使用流式细胞仪。为了满足日益增长的分类需求，多 年实施了战略收购，有效地将 Core 的分选仪器增加了一倍 2017 年（2017 年、2018 年和 2021 年添加了工具）。目前，核心区的所有四台分拣机均得到高度利用率。 三个分选机是 BD Biosciences Influx 仪器，采用空气喷射流体设计和高速、多 孔板沉积力学。这些涌入者对于许多索尔克来说都是多才多艺且可靠的主力 多年来的项目。不幸的是，Influx 系统已经达到了其使用寿命的终点，核心的 单位平均使用年限约为 10 年。代价高昂的实验失败、获得技术支持的机会减少以及 缺乏更换零件的解决方案集体加剧了迁移到更新技术的紧迫性， 特别是索尔克的分拣需求预计将继续增加。此外，两个流入不 符合现行气溶胶遏制标准。此项拨款申请的目的是解决 迫切需要：通过购买先进的现代细胞分选仪来升级Core的分选能力。 Core 提议购买 Thermo Fisher Spectral Bigfoot（5 激光、48 荧光探测器）。这 新仪器将补充当前的分拣能力，并且在由于意外延误而导致的情况下具有无价的价值 技术故障。 Bigfoot 是一流的“空中喷射”型细胞分选机，是替代细胞分选机的唯一可行选择 过时的喷射空气流入系统。无需额外配件：Bigfoot 具有集成的 II 类 生物防护外壳、气溶胶防护功能以及用于板的高速多孔沉积系统 排序。拟议的仪器将使用“光谱”配置，其中有 5 条常见激光线，分别是 用于索尔克表型分析面板。这种 5 激光光谱配置将使研究人员能够利用 先前优化的染色面板，并将添加目前不可用的强大“光谱检测”功能 在索尔克。光谱检测能力的一个主要优点是它可以获取荧光团组合 以前不可能。这反过来又允许将其他感兴趣的抗原轻松添加到现有的组中。 总而言之，利用光谱检测具有显着的成本和时间优势：更多信息 可以从有限的珍贵样本中收集，从而加速科学发现。索尔克研究所有 对实施、支持和长期管理的承诺的良好记录 的高端仪器仪表。凭借索尔克流式细胞术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。