Bigfoot Spectral Cell Sorter

这项请求是为了获得资金购买一台包含生物的ThermoFisher BigFoot光谱分类器(7台激光，60台探测器)。 匹兹堡大学或方圆100英里内都没有全光谱细胞分选器。作为一种独特的乐器， 大脚分类器结合了高参数、高内容成像(多激光、高探测器)的优势 使用现代全光谱技术(算法光谱分解)。我们正在进行的由NIH资助的研究是 受阻于无法直接从皮特的三个光谱中转换复杂的、经过验证的面板 将分析器转换为同等的光谱单元分选器。在光谱方法中，发射的光的全光谱 每个细胞由多个检测器收集，然后进行分解，将信号分离为单独的标记。 光谱细胞术通过允许使用新的荧光和新的组合来扩展参数空间 先前不能在常规配置的细胞仪中配对在一起的现有荧光(即 探测器，一种颜色系统)。PIT的所有分拣机都是传统的配置型。其后果是 我们大部分或所有高度复杂的免疫表型光谱面板都必须缩小规模，以适应 更少的探测器，并重新验证与现有传统分类器的兼容性，所有这些都无法捕获 最初的复杂性和新的信息诱人地揭示了光谱分析仪。此外，我们目前的 传统的分拣机被大量使用。大脚怪的一个非常吸引人的特点是它有两个可操作的 模式，光谱模式和常规模式。因此，大脚怪满足了频谱复杂度高的用户的需求 分拣需求，也使我们非常繁忙的核心的所有用户受益，他们有传统的补偿需求。 所需的BigFoot光谱分选器将安装在皮特的中央统一流式细胞仪核心内 医学院。核心为180多个PI和500个个人用户提供服务。对于提议的乐器，少校 用户包括10个PI(9个由NIH资助)，另外7个PI是次要用户。这些PI总共拥有超过30美元的收入 NIH资助的直接成本至少来自5个不同的研究所。一个主要的研究焦点是在广泛的 免疫学和相关免疫疾病，与移植、皮肤病、血管等姊妹学科 生物学、传染病和肿瘤学也得到了很好的代表。我们的初步和演示工作向我们证明了 这一仪器将真正让我们的研究人员在他们的研究中开辟新的天地。大脚怪将会是 受到统一流式细胞仪核心工作人员的良好照顾，该核心拥有广泛的基础设施和文化 支持面板设计，监督仪器运行，并确保财务自给自足。 除了提供独特和急需的光谱分选能力外，该仪器还将容纳 对可配置分拣的需求，包括6路分拣到管中和直接分拣到1 Ox芯片中，并允许 更好地检测和高效地分类更罕见的事件。该分拣机有一个集成的II类生物安全柜。这个 Required BigFoot将在PIT提供第一台最先进的光谱分选器仪器，以满足 我们的用户基础和先进的研究项目的高复杂性、高内容的需求。

This request is for funds to purchase a biocontained ThermoFisher Bigfoot spectral sorter (7 laser, 60 detector). There is no full-spectrum cell sorter at the University of Pittsburgh or within 100 miles. Unique as an instrument, the Bigfoot sorter combines the strengths of high-parameter, high-content imaging (multi-laser, high-detector) with modern full-spectrum technologies (algorithmic spectral unmixing). Our ongoing NIH-funded research is hampered by the inability to directly transition complex, validated panels from Pitt's three spectral analyzers to an equivalent spectral cell sorter. In spectral approaches, the full spectrum of light emitted from each cell is collected by multiple detectors followed by unmixing to separate the signal into individual markers. Spectral cytometry expands parameter space by permitting the use of new fluors and also new combinations of existing fluors that could not previously be paired together in conventionally configured cytometers (i.e., one detector, one color systems). All of the sorters at Pitt are the conventional configuration type. The consequence is that most or all of our highly complex immunophenotypic spectral panels have to be scaled down, adapted to fewer detectors, and re-validated for compatibility with existing conventional sorters, all of which fails to capture the original complexity and novel information tantalizingly revealed by the spectral analyzers. Further, our current conventional sorters are heavily used. A very attractive feature of the Bigfoot is that it that has two operational modes, spectral and conventional. Thus, the Bigfoot meets the demands of users with high-complexity spectral sorting needs, and also benefits all users of our very busy core who have conventionally compensated needs. The requested Bigfoot spectral sorter will be housed within the centralized Unified Flow Cytometry Core in Pitt's School of Medicine. The core serves over 180 Pis and 500 individual users. For the proposed instrument, Major Users include 10 Pis (9 NIH funded) with another 7 Pis as Minor Users. Together, these Pis have over $30 Min NIH-funded direct costs from at least 5 different institutes. A major research focus is in the broad area of immunology and allied immune diseases, with sister disciplines such as transplantation, dermatology, vascular biology, infectious disease, and oncology also well represented. Our preliminary and demo work proved to us that this instrument will truly allow our investigators to break new ground in their research. The Bigfoot will be well cared for by the staff of the Unified Flow Cytometry Core, which has an extensive infrastructure and culture in place to support panel design, oversee instrument operations, and ensure that it is financially self-supporting. In addition to providing unique and much needed spectral sorting capabilities, the instrument will accommodate demands for configurable sorts including 6-way sorts into tubes and direct sorting into 1 Ox chips, and allow for better detection and efficient sorting of rarer events. The sorter has an integrated Class II biosafety cabinet. The requested Bigfoot will provide the first state-of-the-art spectral sorter instrumentation at Pitt to meet the high-complexity, high-content needs of our user base and advancing research programs.