ShEEP Request for a Replacement Cell Sorting Flow Cytometer for the Flow Cytometry Core Facility

ShEEP 请求为流式细胞术核心设施更换细胞分选流式细胞仪

• 批准号: 9793614
• 负责人: CHRISTINE LINDA HSIEH
• 金额: --
• 依托单位: VETERANS AFFAIRS MED CTR SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-01-01 至 2019-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9793614
• 关键词: Aerosols; Affect; Area; Biological Assay; Biological Sciences; Cardiology; Cell Separation; Cells; Clinical; Color; Communicable Diseases; Communities; Commuting; Computer software; Core Facility; Coupled; Cytometry; Dementia; Detection; Disease; Equipment; Exposure to; Fees; Flow Cytometry; Fluorescence; Foundations; Funding; Future; Gene Expression; Gene Expression Profiling; Generations; Goals; Health; Health Services Research; Heart Injuries; Infectious Agent; International; Lasers; Liquid substance; Maintenance; Malignant neoplasm of prostate; Manufacturer Name; Mind; Minor; Modernization; Molecular and Cellular Biology; Mus; Optics; Pathology; Performance; Population; Principal Investigator; Procedures; Pulmonology; Reagent; Reporter; Research; Research Personnel; Research Project Grants; Rheumatology; Risk; Safety; Sampling; San Francisco; Services; Sheep; Sorting - Cell Movement; Special Equipment; Stains; System; Technology; Temperature; Training; Traumatic Brain Injury; Urology; Veterans; Viola; Work; aged; base; design; detector; exosome; experimental study; high risk population; high throughput analysis; improved; in vivo; innovation; instrument; interest; microvesicles; migration; neurosurgery; next generation; particle; particle detector; protein expression; single cell analysis; therapy design; tool; transcriptome sequencing

Flow cytometry is an outstanding research tool to research molecular and cellular biology. This fluorescence-based technology allows single-cell analysis of a high number of parameters to detect gene and protein expression. A highly valuable aspect of flow cytometry is the ability to sort for multiple specific cell populations, which is a critical step in modern day next generation RNA sequencing experiments and single cell gene expression analyses. The San Francisco VA Flow Cytometry Core Facility supports VA funded research and the VA research community. Multiple departments, including Rheumatology, Pathology, Neurosurgery, Urology, Cardiology and Pulmonology, require the flow cytometry services at this core facility and cell sorting is an integral part of several research projects. For the past 14 years to the present, the most valuable and main component of the core facility is a cell sorting flow cytometer, the Bectin Dickinson (BD) Biosciences FACS Aria IIu. The current cell sorter was manufactured in 2004, and unfortunately, in 2020 it will no longer be supported by the manufacturer; they will discontinue supplying parts and no longer offer service options which is required to maintain our aged equipment. There is a dire need for replacing the core facility’s cell sorter. The replacement equipment requested is an upgraded and advanced version of the current equipment, and it is called the BD Biosciences FACS Aria Fusion, which we will call the Aria Fusion. We expect the migration to the Aria Fusion to be a smooth transition with minimal additional training for management and for users as it utilizes identical software and fluidics mechanisms as the current Aria IIu. The Aria Fusion offers many technological and safety advancements over our current set up that will enhance VA research and increase the number of users. There are four main advancements of the requested flow cytometer: 1) The addition of the yellow-green laser (to replace our current near UV laser) will immensely improve the selection, detection, and the sensitivity of a vast number of current and newly developed fluors. This includes the ability to detect mCherry and tdTomato, popular fluors used in reporter mice. 2) The addition of a manufacturer-designed biosafety cabinet to encase the replacement cytometer will reduce the risk of exposure to infectious agents derived from BSL2 or higher risk group samples that become airborne during sorting. The core’s standard operating procedures (SOP) will be able to meet new international standards of biosafety for cytometry-based cell sorting. This will also expand the number of users to attract research on infectious diseases and clinically-derived samples. 3) The replacement Aria Fusion will have a high-powered blue laser coupled with a small particle detector. This special equipment is required for analyzing microvesicles, such as exosomes. Exosome research is a major interest for many San Francisco VA-funded Principal Investigators, and the new capability will attract new users and expand the research of current users. 4) The Aria Fusion will allow cell sorting into 96 well and 384 well plates. This new feature will facilitate research requiring the precision of single cell gene expression analyses. We are excited to request the Aria Fusion, a state of the art cell sorter that will become the foundation of the core facility and attract new users. Usage fees from all users will be sufficient for maintenance. This replacement is much needed; the current Aria will not be supported in 2020 and it lacks current day safety standards. The Aria Fusion advancements will expand the number of users. We expect the Aria Fusion to provide services critical towards accomplishing VA’s research goals towards understanding and treating diseases of high importance in the Veteran population.

流式细胞术是研究分子和细胞生物学的一种优秀的研究工具。这 基于荧光的技术允许单细胞分析大量的参数来检测 基因和蛋白质的表达。流式细胞术的一个非常有价值的方面是能够对多个 特定的细胞群体，这是现代下一代RNA测序的关键步骤 实验和单细胞基因表达分析。旧金山退伍军人事务部流式细胞术核心 设施支持退伍军人事务部资助的研究和退伍军人事务部研究社区。多个部门，包括 风湿科、病理学、神经外科、泌尿科、心脏科和肺科，需要流动 这一核心设施的细胞学服务和细胞分类是几个研究项目不可或缺的一部分。 从过去14年到现在，核心设施最有价值和最主要的组成部分 是一种细胞分选型流式细胞仪，是Bectin Dickinson(BD)Biosciences FACS Aria IIU。当前单元格 分类器是在2004年生产的，不幸的是，到2020年，它将不再受 制造商；他们将停止供应部件，不再提供所需的服务选项 来维护我们陈旧的设备。迫切需要更换核心设施的细胞分选器。这个 要求更换的设备是当前设备的升级和高级版本，以及 它被称为BD生物科学FACS Aria Fusion，我们将称之为Aria Fusion。我们期待着 迁移到Aria Fusion将平稳过渡，只需最少的额外管理培训 和用户，因为它使用与目前的Aria IIU相同的软件和流体机制。 Aria Fusion比我们目前的设置提供了许多技术和安全方面的进步 这将加强退伍军人管理局的研究，增加用户数量。有四个主要的进步 所要求的流式细胞仪： 1)增加黄绿光激光器(以取代我们目前的近紫外光激光器)将极大地 提高了对当前和新开发的广大仪器的选择、检测和灵敏度 胡说八道。这包括检测mCherry和tdTomato的能力，这是在报告老鼠中使用的流行氟。 2)增加了制造商设计的生物安全柜以包装更换 细胞仪将降低接触BSL2或更高风险人群的感染性病原体的风险 在分拣过程中变得空气中的样本。核心的标准操作程序(SOP)将是 能够满足基于细胞仪的细胞分选的新的国际生物安全标准。这也将是 扩大用户数量，以吸引传染病和临床衍生样本的研究。 3)替代的Aria Fusion将拥有一个高功率的蓝色激光器和一个小粒子 探测器。这种特殊的设备是分析微囊所必需的，例如外切体。外切体 研究是许多旧金山退伍军人事务部资助的首席调查人员的主要兴趣所在，新的 能力将吸引新用户，并扩大对现有用户的研究。 4)Aria Fusion将允许细胞分选到96孔板和384孔板。这一新功能将 促进需要单细胞基因表达分析精确度的研究。 我们很兴奋地要求Aria Fusion，这是一种最先进的细胞分选器，将成为 建立核心设施，吸引新用户。所有用户的使用费将足以满足 维修。这种替代是非常需要的；当前的Aria将在2020年不受支持，而且它 缺乏当今的安全标准。Aria Fusion的进步将扩大用户数量。 我们预计Aria Fusion将提供对实现退伍军人管理局的研究目标至关重要的服务 了解和治疗退伍军人群体中高度重要的疾病。