Spectral Configured Bigfoot Sorter for Salk Institute Flow Cytometry Core

项目摘要 索尔克生物研究所是一个独立的非营利基础科学研究机构，位于 在加利福尼亚州的拉霍亚。索尔克的研究项目包括癌症生物学、免疫学和微生物发病机理、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 技术核心为这项研究提供了关键支持。流式细胞术核心为研究所提供 研究人员获得流式细胞仪仪器。为了满足日益增长的分拣需求，多 年的战略收购已经实施，有效地增加了一倍的核心的分选仪器，因为 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。 三个分选机是BD Biosciences Influx仪器，其特点是空气射流设计和高速，多功能， 孔板沉积力学。这些流入者是许多索尔克人的多才多艺和可靠的主力 多年来的项目。不幸的是，Influx系统已经到了使用寿命的尽头， 平均年龄约为10岁。代价高昂的实验失败，减少获得技术支持，以及 缺乏替换部件的解决方案已经共同地升级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，两个流入者不 符合当前的气溶胶控制标准。本补助金申请的目的是解决一个 迫切需要：通过购买先进的现代细胞分选机来升级Core的分选能力。 核心计划获得一个Thermo Fisher Spectral Bigfoot（5激光，48荧光探测器）。的 新仪器将补充目前的分拣能力，在由于以下原因而出现意外延误时， 技术故障。大脚是一流的“喷气式”细胞分选机，是唯一可行的选择，以取代 过时的空中喷射系统不需要额外的配件：大脚有一个集成的II类 生物防护罩、气溶胶防护特征和用于板的高速多孔沉积系统 分类拟议的仪器将使用一个“光谱”配置，具有5个常见的激光线， 用于Salk表型分析板。这种5激光光谱配置将允许研究人员利用 先前优化的染色面板，并将增加目前不可用的强大的“光谱检测”功能 在索尔克光谱检测能力的一个主要优点是它提供了对荧光团组合的访问 以前不可能。这又允许将另外的感兴趣的抗原容易地添加到现有的组中。 总之，利用光谱检测具有显著的成本和时间优势：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 对实施、支持和长期管理的承诺的良好记录 高端仪器仪表。凭借Salk流式细胞仪核心提供的经验和专业知识， 新仪器将立即在高级别上使用，从而促进研究所的科学发现。

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.