ShEEP Request for 5 Laser 28 parameter Flow Cytometer

这项设备的应用是为了升级我们在克利夫兰退伍军人医学中心的流式细胞仪能力。 目前，3个主要实验室(Anthony、Canaday、Shive)在过去6年中受益于2014年的BD LSRFortessa 5激光(蓝、红、紫、黄/绿、紫外线)16色，流式细胞仪。我们现在有了一个 增加VA资助的大容量流式细胞仪用户(本申请中的7个主要用户，包括 Anthony、Canaday、Shive、Wald、King、Hise、Zidar，预计2020年将增加1-2个新用户)。 拟议的A5 SYMPHONY 5激光/28检测器(彩色)配置流式细胞仪将允许用户 换成参数更高的高端仪器。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，使目前的用户可以轻松过渡到。 2)A5有5个激光器，但随着可用的荧光染料的扩展，可以升级到9个激光器。 3)A5拥有更高的探测器能力，使目前的16色限制增加到最高28色 没有更多激光的限制 4)更快的处理器 5)更好地优化了每个探测器的PMT 拟议的最终结果将实现两个主要目标： 1)允许保存细胞资源，允许通过以下方式对给定体积的血液进行更多研究 将流板组合成一根管子。例如，具有28种颜色的单个流面板可以 提供有关B细胞、树突状细胞和单核细胞的信息，为深低温保存的外周血细胞进行优化 血液单个核细胞。能够在有限的血液容量上实现最高数量的研究 细胞数量在新冠肺炎研究中尤为重要，因为实际细胞产量降低了 在这些患者中，由于他们的危重疾病和 我们想要的纵向抽样。 2)允许更高的表型分辨率 多个描述良好的28个彩色流动面板(已发布和在中建议的)可以提供 在分析T细胞、iNKT和NK、树突状细胞和检查点方面具有显著的分析深度 路径聚焦。 拟议的BD A5交响乐将包括： 5激光/28色A5 FACS5激光(蓝宝石488-200、OBIS637-140、紫罗兰405-200、OBIS 561-150，UV 355-60)流式细胞仪(允许28种颜色) 高吞吐量系统(HTS) HP Z2G4 Win10，带34英寸显示器 用于BV510和V450的Extra滤光片 荧光染料及原型染料评价试剂盒 HP面板设计入门套件(50个小瓶装，包括现成的、定制的或可选的) FACS流量供给系统 量子闪光发光二极管脉冲星

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