ShEEP Request for 5 Laser 28 parameter Flow Cytometer

ShEEP 请求 5 激光 28 参数流式细胞仪

• 批准号: 10176764
• 负责人: Donald D Anthony
• 金额: --
• 依托单位: LOUIS STOKES CLEVELAND VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-10-01 至 2021-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10176764
• 关键词: Algorithms; B-Lymphocytes; Blood Cell Count; Blood Volume; COVID-19; Cell physiology; Cell surface; Cells; Cellular Immunology; Clinical Pathology; Color; Complex; Computer software; Contract Services; Critical Illness; Cryopreservation; Custom; Dendritic Cells; Dyes; Equipment; Evaluation; Fees; Flow Cytometry; Fluorochrome; Funding; Goals; Grant; Human; Laboratories; Lasers; Maintenance; Medical center; Methodology; Monitor; Mus; Optics; Pathway interactions; Patients; Peripheral Blood Mononuclear Cell; Phenotype; Publishing; Pulsar; Research; Resolution; Resources; Sampling; Sapphire; Schedule; Services; Sheep; System; T-Lymphocyte; Technology; Tube; Vial device; Viola; Wing; base; cost; design; detector; immunoregulation; instrument; monocyte; operation; prototype

This equipment application is to upgrade our Flow Cytometry capability at the Cleveland VA Medical center. Currently 3 main laboratories (Anthony, Canaday, Shive) have benefitted over the past 6 years from a 2014 BD LSRFortessa 5 laser (Blue, Red, Violet, Yellow/Green, UV) 16 color, Flow Cytometer. We now have an increase in VA funded large volume flow cytometry users (7 main users in present application, including Anthony, Canaday, Shive, Wald, King, Hise, Zidar and with 1-2 additional incoming users anticipated in 2020). The proposed A5 Symphony 5 Laser/ 28 detector (color) configuration flow cytometer will allow the users to move to a higher parameter top end instrument. Symphony advantages include: 1) highest performing commercial fluorochrome based flow cytometer, with similar operation parameters and interfacing software so that current users can transition to easily. 2) A5 comes with 5 lasers, but can be upgraded to 9 lasers as available fluorochromes extend. 3) A5 has higher detector capability, making for increase from current 16 color limit to up to 28 color limit without more lasers 4) Faster processors 5) Better optimized PMT for each detector The end-result of the proposed will accomplish 2 main goals: 1) Allow for cell resource conservation allowing more studies on a given volume of blood by combining flow panels into a single tube. As an example, a single flow panel with 28-colors can provide information on B cells, dendritic cells, and monocytes, optimized for cryopreserved peripheral blood mononuclear cells. Being able to achieve the highest number of studies on limited blood volumes and cell numbers is particularly important in the COVID-19 studies since actual cell yields are reduced in these patients and achievable blood volumes are also diminished due to their critical illness and the longitudinal sampling that we desire. 2) Allow for higher phenotype resolution A number of well described 28 color flow panels (published, and ones proposed within) can provide remarkable depth of analysis for analysis of T cells, iNKT and NK, dendritic cells, and checkpoint pathway focus. The proposed BD A5 Symphony will include:  5 Laser/ 28 Color A5 FACS Symphony 5 laser (Sapphire 488-200, OBIS 637-140, Violet 405-200, OBIS 561-150, UV 355-60) Flow Cytometer (allowing 28 colors)  High Throughput System (HTS)  HP Z2G4 Win10 w/ 34” monitor  Extra optical filters for BV510 and V450  Fluorochrome & Prototype Dye evaluation kit  HP Panel Design Starter Kit (50 small vials which included off the shelf, custom, or Optibuild)  FACS Flow Supply System  Quantiflash LED Pulsar

该设备应用旨在升级克利夫兰VA医疗中心的流式细胞术能力。 目前，3个主要实验室（Anthony、Canadian、Shive）在过去6年中受益于2014年BD LSRFortessa 5激光（蓝色、红色、紫色、黄色/绿色、UV）16色，流式细胞仪。我们现在有一个 VA资助的大容量流式细胞仪用户增加（本申请中有7个主要用户，包括 Anthony、Canadian、Shive、Wald、King、Hise、Zidar，预计2020年将有1-2名新用户加入）。 申报的A5 Symphony 5激光/ 28检测器（彩色）配置流式细胞仪将允许用户 转向更高参数的高端仪器。Symphony的优势包括： 1)性能最高的商用荧光染料流式细胞仪，具有类似的操作 参数和接口软件，以便当前用户可以轻松过渡到。 2)A5配备5个激光器，但可以升级到9个激光器作为可用的荧光染料扩展。 3)A5具有更高的探测器能力，使其从当前的16色限制增加到28色 没有更多激光器的极限 4)更快的处理器 5)为每个探测器优化PMT 拟议的最终结果将实现两个主要目标： 1)允许细胞资源保存，允许对给定体积的血液进行更多研究， 将流动面板组合到单个管中。例如，具有28种颜色的单个流面板可以 提供B细胞、树突状细胞和单核细胞的信息， 血单核细胞能够在有限的血量上实现最多的研究 细胞数量在COVID-19研究中尤为重要，因为实际细胞产量会减少 在这些患者中，可获得的血量也由于他们的危重疾病而减少， 我们想要的纵向采样。 2)允许更高的表型分辨率 许多良好描述的28色流面板（已公布，以及其中提出的）可以提供 T细胞、iNKT和NK、树突状细胞和检查点分析的显著分析深度 路径聚焦 拟议的BD A5 Symphony将包括： 105激光/ 28色A5 FACS Symphony 5激光（Sapphire 488-200、OBIS 637-140、Violet 405-200、OBIS 561-150，UV 355-60）流式细胞仪（允许28种颜色） 高温超导系统（HTS） HP Z2 G4 Win10 w/ 34”显示器 适用于BV 510和V450的额外滤光片 荧光染料和原型染料评估套件 HP Panel Design Starter Kit（50小瓶，包括现成的、定制的或Optibuild） 流体供应系统 Quantiflash LED Pulsar