ShEEP Request for Fusion Flow Cytometer and Cell Sorter

ShEEP 请求融合流式细胞仪和细胞分选仪

• 批准号: 9210823
• 负责人: Shekar N. Kurpad
• 金额: --
• 依托单位: CLEMENT J. ZABLOCKI VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-05-01 至 2016-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9210823
• 关键词: Address; Adult; Aerosols; Animals; Area; Arthritis; Bacteria; Biological; Biological Markers; Blood; Bronchoalveolar Lavage; Calcium pyrophosphate deposition disease; Cell Culture Techniques; Cell Cycle; Cell Death; Cell Separation; Cell Survival; Cell Therapy; Cell surface; Cells; Characteristics; Chondrocytes; Collaborations; Collection; Complement; Computer Workstations; Computer software; Contract Services; Core Facility; Degenerative polyarthritis; Detection; Development; Devices; Diffusion Magnetic Resonance Imaging; Dyes; Ensure; Environment; Environmental air flow; Enzyme-Linked Immunosorbent Assay; Equipment; Excision; Exposure to; Face; Fee-for-Service Plans; Fees; Flow Cytometry; Funding; General Population; Goals; Grant; HIV-1; Healthcare; Housing; Human; Human T-lymphotropic virus 2; Image; Image Analysis; Immune response; In Vitro; Inflammation; Institution; Ischemia; Journals; Label; Laboratories; Lasers; Lesion; Life; Location; Lung Transplantation; Magnetic Resonance Imaging; Maintenance; Malignant Epithelial Cell; Marketing; Medical center; Mesenchymal Stem Cells; Methods; Microscope; Mission; Mitochondria; Modeling; Molecular; Oxidative Stress; Oxidative Stress Induction; Performance; Play; Population; Power Sources; Preparation; Procedures; Production; Property; Proteins; Public Health; Publishing; Rattus; Reactive Oxygen Species; Reader; Rectal Cancer; Reperfusion Injury; Reperfusion Therapy; Research; Research Personnel; Research Project Grants; Resources; Risk; Role; Safety; Sample Size; Sampling; Serotonin Receptors 5-HT4; Signal Transduction; Site; Sorting - Cell Movement; Source; Spinal cord injury; Staining method; Stains; Sterility; Structure of parenchyma of lung; Surface; System; Therapeutic Intervention; Time; Tissues; Training; Transitional Cell Carcinoma; Translating; Transport Process; Traumatic Brain Injury; Travel; Veterans; Viral Physiology; Virus; Western Blotting; Wisconsin; Work; antibody conjugate; base; chemokine; cost; cytokine; cytotoxicity; density; fluorophore; improved; in vivo; injured; instrument; lung injury; lung ischemia; medical schools; prevent; programs; regenerative; research study; tool; treatment effect

The primary justification for the requested system is the current lack of flow cytometry and cell sorting at the Milwaukee VA despite a high demand for such procedures. The requested system is a BD FACSARIA Fusion Special Order flow cytometry and cell sorting system with all components necessary to execute multicolor flow cytometry and cell sorting experiments. The instrument will consist of five lasers, which allows for detection of 18 fluorescent parameters. This will enable researchers at the Milwaukee VA to detect a large variety of cellular markers in the same cell. This instrument is produced by the market leader BD. VA researchers have previously used flow cytometry and cell sorting at the core facilities of the Medical College of Wisconsin. The same instruments and software are represented at the core facilities, which would allow researchers at the VA to continue their experiments without technical adjustment. The research projects which would strongly benefit from the presence of a flow cytometer and cell sorter at the VA include a variety of conditions relevant to both Veterans and the general population. Dr. Kurpad is using diffusion tensor imaging (DTI) analysis of the injured spinal cord at the lesion site and proximal to the lesion. Flow cytometry will be used to verify imaging results and to identify and correlate cell activation, cell death and cytotoxicity to the DTI results. Dr. Jacobs is investigating ischemia-reperfusion lung injuries, a commonly encountered condition after lung transplantation. Flow cytometry will be performed on cells derived from bronchoalveolar lavage and on enzymatically digested lung tissue and will provide information about cell signaling and mitochondrial properties. Furthermore, the result of therapeutic interventions will be assessed by flow cytometry. Dr. Rosenthal's research is focused on a form of degenerative arthritis, the calcium pyrophosphate deposition disease. The cell sorter would be used to sort populations of successfully transfected chondrocytes which can then be used for further experiments. Dr. See's project addresses the induction of oxidative stress in urothelial carcinoma cells by BCG bacteria. This oxidative stress is a critical component for the antitumor activity of BCG. Flow cytometry will be used to detect treatment effects in terms of cytotoxicity and cell death. Dr. Kroner-Milsch is investigating the role of inflammation after spinal cord injury (SCI). She aims to reduce secondary damage by specifically blocking cytokines and chemokines which are upregulated following SCI. Cells derived from injured tissue will be analyzed for production of cytokines and reactive oxygen species. Dr. Glavaski-Joksimovic's research focus is the neuroprotective effect of adult mesenchymal stem cells (MSC) after traumatic brain injury (TBI). Flow cytometry and cell sorting will be used to study and sort different MSC subpopulations and compare their effects in TBI models. Dr. Beilke is investigating the role of HTLV-2 virus as a modulator of immune response. Tax2, a HTLV-2 protein, is a strong chemokine regulator and plays a role in anti-viral activity against HIV-1. Flow cytometry is frequently applied in his studies to detect expression of surface molecules in specific cell populations. Dr. Budde's research focus is the development of noninvasive MRI biomarkers for SCI and TBI. A flow cytometry and cell sorting instrument will be necessary for cell based therapy studies with a high promise of neuroprotective and regenerative effects. Dr. Ridolfi investigates the role of 5-HT3 and 5-HT4 receptors after rectal cancer resection. Flow cytometry will be used to assess expression density in his rat model of parasympathetic damage. In summary, access to a flow cytometer and cell sorter at the Milwaukee VA would accelerate research projects currently done at the VA and open new research opportunities for VA investigators.

所需系统的主要理由是目前缺乏流式细胞仪和细胞分类 弗吉尼亚州密尔沃基，尽管对这类程序的需求很高。 所要求的系统是BD FACSARIA Fusion特殊顺序流式细胞仪和细胞分选系统 进行多色流式细胞术和细胞分选实验所需的所有组件。这台仪器将 由五个激光器组成，可检测18个荧光参数。这将使研究人员能够 密尔沃基退伍军人管理局在同一细胞中检测到多种细胞标志物。这台仪器是由 市场领头羊BD。退伍军人管理局的研究人员此前在核心设施中使用了流式细胞术和细胞分选 威斯康星医学院的。在核心设施中代表相同的仪器和软件， 这将允许退伍军人管理局的研究人员在不进行技术调整的情况下继续他们的实验。 将从流式细胞仪和细胞分选器的存在中受益的研究项目 退伍军人包括各种与退伍军人和普通民众相关的疾病。 库尔帕德博士正在使用扩散张量成像(DTI)分析损伤脊髓的病变部位和 在病变的近端。流式细胞术将用于验证成像结果，并识别和关联细胞 激活、细胞死亡和细胞毒性对DTI的结果。 雅各布斯博士正在研究缺血再灌注肺损伤，这是继肺之后的一种常见情况。 移植。流式细胞术将对来自支气管肺泡灌洗和其他来源的细胞进行检测 并将提供有关细胞信号和线粒体特性的信息。 此外，治疗干预的结果将通过流式细胞仪进行评估。 罗森塔尔博士的研究重点是一种形式的退行性关节炎，即焦磷酸钙沉积 疾病。该细胞分选器将用于对成功转基因的软骨细胞群体进行分选，这可以 然后用于进一步的实验。 See博士的项目研究了卡介苗细菌对尿路上皮癌细胞的氧化应激诱导作用。 这种氧化应激是卡介苗抗肿瘤活性的关键成分。流式细胞术将用于 根据细胞毒性和细胞死亡检测治疗效果。 Kroner-Milsch博士正在研究脊髓损伤(SCI)后炎症的作用。她的目标是减少 通过特异性地阻断脊髓损伤后上调的细胞因子和趋化因子而造成的二次损伤。单元格 从受伤组织中提取的物质将被分析以产生细胞因子和活性氧物种。 格拉瓦斯基-约克西莫维奇博士的研究重点是成人间充质干细胞的神经保护作用 (MSC)创伤性脑损伤(TBI)后。将使用流式细胞术和细胞分选来研究和分类不同的 MSC亚群并比较它们在脑损伤模型中的作用。 拜尔克博士正在研究HTLV-2病毒作为免疫反应调节器的作用。Tax2，一种HTLV-2 蛋白质，是一种强大的趋化因子调节因子，在抗艾滋病毒-1的病毒活性中发挥作用。流式细胞术是 在他的研究中经常用于检测特定细胞群体中表面分子的表达。 巴德博士的研究重点是开发脊髓损伤和脑外伤的非侵入性MRI生物标记物。一种流动 细胞仪和细胞分选仪器将是基于细胞的治疗研究所必需的，有很高的希望 神经保护和再生作用。 Ridolfi博士研究了直肠癌切除后5-HT3和5-HT4受体的作用。流式细胞术 将被用来评估他的副交感神经损伤大鼠模型的表达密度。 总而言之，在密尔沃基退伍军人管理局使用流式细胞仪和细胞分选仪将加速研究 目前在退伍军人管理局完成的项目，为退伍军人管理局的调查人员提供了新的研究机会。