ShEEP Request for Fluorescence-Activated Cell Sorter (FACS)

该项目是ShEEP对Becton Dickinson（BD）FACSMelodyTM细胞分选仪的申请。屋宇署 FACSMelodyTM细胞分选仪是一种高质量的流式细胞仪分选仪器，具有自动化 仪器设置例程，具有易于学习和使用的优点。该股提供 研究人员的支持和全面的工作流程经验，并来自引用 BD FACS Aria细胞分选仪通常系统在不到17分钟内就准备好了- 最大限度地提高效率。小单位创建精简的工作流程与仪器设置，以及 作为门和排序设置，可从保存的实验中获取。细胞分选仪 系统预测潜在问题，并确定过滤器/镜像单元的根本原因， 与软件进行通信，以确认光学配置支持实验。的 工具将有助于具有广泛不同项目的各个VA调查人员隔离个体 从正常和疾病组织中提取细胞类型，并研究它们的特征，从而在 对组织炎症、癌症免疫学和神经系统再生的理解。 该仪器的特性使其成为我们VA研究服务的理想选择，因为该奖项 将支持5个主要VA Merit资助用户（Hornyak，Atamas，Luzina，Davila和 Makar）和一名未成年用户（Golding博士），他是职业发展奖（CDA-2）的获得者。 托马斯J. Hornyak，医学博士，哲学博士、是本提案的主要研究者（PI）。他是 弗吉尼亚州马里兰州卫生部研究与开发（R&D）助理参谋长（ACOS） 护理系统（VAMHCS），以及BLR&D Merit Review项目“黑素细胞干细胞在 再生医学”。他和他的实验室将使用分类器分离出两种小鼠的亚群， 和人类皮肤来源的干细胞在活的状态，以评估其性能，以促进神经 再生Sergei Atamas，医学博士，哲学博士、和医学博士伊琳娜·卢齐纳哲学博士、个人资助的VA Merit Review研究者，实验室位于巴尔的摩VA医学中心（VAMC） 他们合作研究白细胞介素（IL）-33和肺纤维化。他们会把组织分类- 衍生的巨噬细胞和淋巴细胞与FACSMelodyTM。Eduardo Davila博士，是一个VA Merit Review研究者，其研究项目专注于肿瘤免疫学。他将使用 细胞分选仪，用于他的VA Merit Review项目，以分离表达不同水平IRAK-4的肿瘤细胞， 假定介导肿瘤对治疗的抗性。塔帕斯·马卡尔博士，是退伍军人管理局的一个优秀资助的 对确定脑源性神经营养因子（BDNF）能力感兴趣的研究者 调节小鼠实验性自身免疫性脑炎， 脱髓鞘疾病他将从小鼠脾脏中分离免疫细胞和炎症细胞 在实验性治疗后评估这些治疗的免疫抑制副作用。 Amit Golding，医学博士，博士是VAMHCS风湿病学家和CDA获奖者，治疗退伍军人， 巴尔的摩VAMC关节炎诊所他提交了VA BLR&D Merit Review申请（秋季 2016年周期）之后，他的CDA-2项目;并有一个研究计划，强调 自身免疫性疾病戈尔丁博士将从血清中分离出B淋巴细胞和T调节细胞 自身免疫性疾病患者和正常对照组的样本进行研究。

The project is a ShEEP request for a Becton Dickinson (BD) FACSMelodyTM Cell Sorter. The BD FACSMelodyTM Cell Sorter is a high quality flow cytometry sorting instrument, with an automated instrument setup routine, that has the advantage of being easy to learn and use. The unit provides researchers a supportive and comprehensive workflow experience, and is derived from the cited and proven BD FACS Aria Cell Sorter. Typically the system is ready in less than 17 minutes- maximizing uptime. The small unit creates streamlined workflow with instrument settings, as well as gates and sort settings available and retrievable from saved experiments. The cell sorter system anticipates potential issues and identifies root causes with filter/mirror units that communicate with the software to confirm the optical configuration supports for experiments. The instrument will facilitate individual VA investigators with widely varying projects to isolate individual types of cells from normal and disease tissue and study their characteristics, leading to advances in the understanding of tissue inflammation, cancer immunology, and nervous system regeneration. The characteristics of the instrument make it ideal for our VA Research Service because this award will support the work of 5 major VA Merit-funded users (Drs. Hornyak, Atamas, Luzina, Davila, and Makar), and a minor user (Dr. Golding) who is a Career Development Award (CDA-2) recipient. Thomas J. Hornyak, M.D., Ph.D., is the Principal Investigator (PI) for this proposal. He is the Associate Chief Of Staff (ACOS) for Research and Development (R&D) at the VA Maryland Health Care System (VAMHCS), and PI on the BLR&D Merit Review project “Melanocyte Stem Cells in Regenerative Medicine”. He and his laboratory will use the sorter to isolate subsets of both mouse and human skin-derived stem cells in the viable state to evaluate their properties to promote neural regeneration. Sergei Atamas, M.D., Ph.D., and Irina Luzina, M.D., Ph.D., are individually funded VA Merit Review investigators with laboratories located in the Baltimore VA Medical Center (VAMC) who collaborate on the study of interleukin (IL)-33 and pulmonary fibrosis. They will sort tissue- derived macrophages and lymphocytes with the FACSMelodyTM. Eduardo Davila, Ph.D., is a VA Merit Review investigator whose research program is focused on tumor immunology. He will use the cell sorter for his VA Merit Review project to isolate tumor cells expressing varying levels of IRAK-4, postulated to mediate tumor resistance to therapy. Tapas Makar, Ph.D., is a VA Merit-funded investigator who is interested in determining the ability of brain-derived neurotrophic factor (BDNF) to modulate murine experimental autoimmune encephalitis, a disease model for autoimmune demyelinating disease in humans. He will sort immune and inflammatory cells from mouse spleen after experimental treatments to evaluate the immunosuppressive side effects of those treatments. Amit Golding, M.D., Ph.D. is a VAMHCS rheumatologist and a CDA awardee that treats Veterans in the Baltimore VAMC Arthritis Clinic. He submitted a VA BLR&D Merit Review application (Fall 2016 Cycle) subsequent to his CDA-2 project; and has a research program emphasizing autoimmune disease. Dr. Golding will sort B lymphocytes and T regulatory cells from serum samples from patients with autoimmune disease and normal controls for his studies.