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术和细胞分选， 密尔沃基弗吉尼亚州，尽管这种程序的需求很高。 申请的系统是BD FACSARIA Fusion Special Order流式细胞术和细胞分选系统， 所有必要的组件来执行流式细胞术和细胞分选实验。仪器将 由五个激光器组成，允许检测18个荧光参数。这将使研究人员在 密尔沃基VA检测同一细胞中的多种细胞标志物。该仪器由 市场领导者BD弗吉尼亚州的研究人员以前曾在核心设施使用流式细胞术和细胞分选 威斯康星州医学院的教授同样的仪器和软件在核心设施中， 这将使退伍军人事务部的研究人员能够在没有技术调整的情况下继续他们的实验。 研究项目将大大受益于流式细胞仪和细胞分选仪的存在， 退伍军人事务部包括与退伍军人和普通民众有关的各种情况。 Kurpad博士正在使用扩散张量成像（DTI）分析损伤部位的受损脊髓， 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 活化、细胞死亡和细胞毒性对DTI结果的影响。 雅各布斯博士正在研究缺血再灌注肺损伤，这是肺损伤后常见的情况。 移植将对来自支气管肺泡灌洗液的细胞和来自支气管肺泡灌洗液的细胞进行流式细胞术。 酶消化的肺组织，并将提供有关细胞信号和线粒体特性的信息。 此外，将通过流式细胞术评估治疗干预的结果。 罗森塔尔博士的研究集中在一种退行性关节炎，焦磷酸钙沉积 疾病细胞分选器将用于分选成功转染的软骨细胞群， 然后用于进一步的实验。 See博士的项目致力于BCG细菌诱导尿路上皮癌细胞的氧化应激。 这种氧化应激是BCG抗肿瘤活性的关键组分。流式细胞术将用于 检测细胞毒性和细胞死亡方面的治疗效果。 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 通过特异性阻断SCI后上调的细胞因子和趋化因子而引起的继发性损伤。细胞 将分析来源于损伤组织的细胞因子和活性氧物质的产生。 Glavaski-Joksimovic博士的研究重点是成体间充质干细胞的神经保护作用 (MSC)创伤性脑损伤（TBI）。流式细胞术和细胞分选将用于研究和分选不同的细胞。 MSC亚群，并比较它们在TBI模型中的作用。 Beilke博士正在研究HTLV-2病毒作为免疫反应调节剂的作用。Tax2，HTLV-2 蛋白，是一种强趋化因子调节剂，在抗HIV-1的抗病毒活性中发挥作用。流式细胞术是 在他的研究中经常应用于检测特定细胞群中表面分子的表达。 Budde博士的研究重点是开发SCI和TBI的非侵入性MRI生物标志物。的流程 细胞计数和细胞分选仪器对于基于细胞的治疗研究是必要的， 神经保护和再生作用。 Ridolfi博士研究了直肠癌切除术后5-HT3和5-HT4受体的作用。流式细胞术 将用于评估他的副交感神经损伤大鼠模型中的表达密度。 总之，在密尔沃基VA获得流式细胞仪和细胞分选仪将加速研究 目前在VA完成的项目，并为VA调查人员打开新的研究机会。