BD LSR II BLUE LASER FLOW CYTOMETER: T CELLS IN GRANULOMATOUS IMMUNE RESPONSES

BD LSR II 蓝色激光流式细胞仪：肉芽肿免疫反应中的 T 细胞

• 批准号: 7335001
• 负责人: Matyas Sandor
• 金额: $ 3.03万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-04-15 至 2007-04-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7335001
• 关键词: DNA; Macaca; Retroviridae; Trypanosoma; brain; cell cycle; cell membrane; cytotoxicity; experience; fasting; flow cytometry; fluorescence; granuloma; immunology; intracellular; lasers; lymph nodes; lymphocyte; macrophage; memory; proteins; tissues

This subproject is one of many research subprojects utilizing the resources provided by a Shared Instrumentation Grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the grant, which is not necessarily the institution for the investigator. DESCRIPTION (provided by applicant): This grant application requests NIH support for the purchase of a BD Biosciences LSR II Blue Laser flow cytometer with four lasers. This very sophisticated piece of equipment will allow our user group to take advantage of the fastest developments in fluorescent technology. Our group of applicants (four major users and three minor users) all work on infectious immunology and rely heavily on cytofluorimetry as a major experimental technology. The LSR II is a highly sensitive and versatile cytometer which will vastly improve the efficiency with which we can measure expression of cell surface and intracellular proteins, assay for in vivo cytotoxicity, and perform DNA cell cycle analysis. This instrument is not currently available on campus for use with BL2/BL3 samples. Use of this instrument will make it possible to detect very rare populations from small sample sizes. Members of our group study very minute cell populations such as brain-infiltrating lymphocytes, memory CDS T cells in nonlymphoid tissue, specialized macrophage subsets, and pathogen-induced single granuloma lesions. In addition, this instrument would allow us to use new fluorescence markers that cannot be detected by our current cytofluorimeters. These new markers are important because they will allow the use of new multicolor transgenic or retrovirus-mediated in vivo labeling of cells and multi-color infectious agents. Finally, all of us are very experienced in 4-color flow cytometry; however, our projects would benefit greatly from being able to measure more parameters from the same samples. The analysis of 10 to 14 parameters in an interdependent way is not currently possible using 4-color cytometry, and is particularly important for both the limiting sample sizes discussed above and samples of experimental rarity, such as lymph nodes or mucosal samples prepared from SIV infected rhesus macaques where we want the most possible information from the least possible number of animals. Analysis of unfixed samples from animals infected with diverse pathogens including Histoplasma and Trypanosoma would be a real advance to our research programs. Advances in fluorescence technology have exploded in the past few years, especially as applied to infectious immunology. A new generation of flow cytometric assays is becoming available and our group of users would like to advance to the next level.

该子项目是利用 NIH/NCRR 资助的共享仪器补助金提供的资源的众多研究子项目之一。子项目和研究者 (PI) 可能已从另一个 NIH 来源获得主要资金，因此可以在其他 CRISP 条目中得到体现。列出的机构是资助机构，不一定是研究者的机构。描述（由申请人提供）：本拨款申请请求 NIH 支持购买具有四个激光器的 BD Biosciences LSR II 蓝色激光流式细胞仪。这种非常先进的设备将使我们的用户组能够利用荧光技术最快的发展。我们的申请人组（四名主要用户和三名次要用户）都从事感染免疫学工作，并严重依赖细胞荧光测定法作为主要实验技术。 LSR II 是一款高度灵敏且多功能的细胞仪，它将极大地提高我们测量细胞表面和细胞内蛋白质表达、测定体内细胞毒性以及进行 DNA 细胞周期分析的效率。该仪器目前在校园内无法用于 BL2/BL3 样本。使用该仪器将可以从小样本量中检测非常稀有的群体。我们小组的成员研究非常微小的细胞群，例如脑浸润淋巴细胞、非淋巴组织中的记忆 CDS T 细胞、专门的巨噬细胞亚群和病原体诱导的单一肉芽肿病变。此外，该仪器将使我们能够使用当前细胞荧光计无法检测到的新荧光标记。这些新标记很重要，因为它们将允许使用新的多色转基因或逆转录病毒介导的细胞和多色感染剂体内标记。最后，我们所有人都对四色流式细胞术非常有经验；然而，如果能够从相同的样本中测量更多参数，我们的项目将会受益匪浅。目前使用 4 色细胞计数法无法以相互依赖的方式对 10 至 14 个参数进行分析，这对于上述讨论的有限样本量和实验稀有样本尤其重要，例如从感染 SIV 的恒河猴制备的淋巴结或粘膜样本，我们希望从尽可能少的动物数量中获得最多可能的信息。对感染组织胞浆菌和锥虫等多种病原体的动物的未固定样本进行分析将是我们研究项目的真正进步。荧光技术在过去几年中取得了爆炸式的进步，尤其是在感染免疫学方面的应用。新一代流式细胞术分析即将上市，我们的用户组希望提升到一个新的水平。