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.

该副本是利用NIH/NCRR资助的共享仪器赠款提供的资源的许多研究子项目之一。子弹和调查员（PI）可能已经从其他NIH来源获得了主要资金，因此可以在其他清晰的条目中代表。列出的机构是赠款，这不一定是调查人员的机构。描述（由申请人提供）：本赠款申请要求NIH支持购买BD Biosciences LSR II蓝色激光流式细胞仪和四个激光。这种非常复杂的设备将使我们的用户组能够利用荧光技术最快的发展。我们的一组申请人（四个主要用户和三个未成年人）都从事传染性免疫学工作，并在很大程度上依赖于细胞氟化法作为一项主要的实验技术。 LSR II是一种高度敏感且通用的细胞图仪，它将大大提高我们可以测量细胞表面和细胞内蛋白的表达的效率，用于体内细胞毒性的测定，并执行DNA细胞周期分析。该乐器目前尚未在校园内与BL2/BL3样品一起使用。该仪器的使用将使从小样本量中检测出非常罕见的种群是可能的。我们小组的成员研究了非常小的细胞群体，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以及病原体诱导的单肉芽肿病变。此外，该仪器将使我们能够使用我们当前的细胞荧光仪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的体内标记。最后，我们所有人在四色流式细胞仪中都非常有经验。但是，我们的项目将从相同样本中衡量更多参数中受益匪浅。目前无法使用4色细胞术以相互依存的方式对10至14个参数进行分析，对于上面讨论的限制样本量和实验稀有性样本，尤其重要，例如淋巴结或粘膜样品，例如由SIV感染的鼠尾草中的淋巴结样本或粘膜样本，我们希望我们希望从最不可能的动物数量中获得最大可能的信息。对感染包括组织肿瘤和锥虫（包括组织型病原体）感染的动物的未固定样品的分析将是我们研究计划的真正进步。在过去的几年中，荧光技术的进步爆炸了，尤其是应用于传染性免疫学。新一代的流式细胞仪测定正在获得，我们的一组用户希望提高到一个新的水平。