RESEARCH FLOW CYTOMETER

与传统的显微荧光法或放射自显影技术相比，流式细胞术具有显著的优势。首先，流式细胞术可以快速分析单个细胞或细胞群体内的分布。通过这种方式，可以分析给定种群中的大量细胞，甚至可以检测到最罕见的亚群。具有越来越高的特异性和报告能力的荧光染料的出现允许同时测量细胞群体内或不同细胞群体之间的多个参数。随着染色技术和条件的进步，更好地保持细胞活性和膜完整性，使得用流式细胞术对细胞周期和抗原表达进行详细分析成为可能。流式细胞术与细胞分选的互补提供了额外的优势，即一旦细胞被荧光鉴定为具有不寻常或所需的特征，就可以物理分离细胞以供进一步研究。这项建议是基于迫切需要利用流式细胞术的这些优势来实现正在进行的NIH资助的研究的特定目标。研究人员将使用流式细胞术技术来实现这些目标，其中包括：阐明抗菌肽之间的结构/功能关系；表征对抗菌剂敏感或耐药的细胞；筛选核酸表达文库；确定病原体、内皮细胞和中性粒细胞之间的相互作用；以及通过产生细胞因子或表达白细胞黏附分子来鉴定对免疫调节刺激特异反应的细胞。获得拟议的文书将大大推进这些研究的进展并扩大其范围。因此，我们确定了在港湾-加州大学洛杉矶分校医学中心园区建立这一研究流式细胞仪的三个主要目标：i)为校园内的多名研究人员提供可访问的、经济的专用研究流式细胞仪；ii)协调灵活的网络，使研究人员能够使用荧光染料和根据其特定应用定制的条件进行流式细胞仪分析；以及iii)为使用现代流式细胞仪技术经验较少的研究人员推广技术专长。这些目标将使调查人员能够实现正在进行的NIH资助项目的具体目标，并生成向NIH和其他机构提交竞争性拨款申请所必需的初步数据。港湾-加州大学洛杉矶分校研究和教育研究所已分配专用空间来容纳该仪器，该机构承诺通过协调充值费用以及为其长期使用提供维护支持和管理费用，为这一重要研究设施的发展提供大力支持。

Flow cytometry offers significant advantages when compared with techniques such as traditional microfluorometry or autoradiography. First, flow cytometry allows for rapid analysis of distribution within single cells or cell populations. In this way, large numbers of cells from a given population can be analyzed, and even the most rare subpopulations detected. The advent of fluorochromes with increasing specificities and reporter capabilities allows for the simultaneous measurement of multiple parameters within a population of cells, or among distinct cell populations. Advances in staining techniques and conditions which better preserve cell viability and membrane integrity have enabled detailed analyses of cell cycle and antigen expression by flow cytometry. Complementation of flow cytometry with cell sorting provides the additional advantage of physical separation of cells for further study once they have been identified by fluorescence to have unusual or desirable characteristics. This proposal is based on the urgent need to capitalize on these advantages of flow cytometry to achieve specific aims of ongoing NIH-funded research. Investigators will use flow cytometry techniques to achieve these goals, which include: elucidation of structure/function relationships among antimicrobial peptides; characterization of cells which are susceptible or resistant to antimicrobial agents; screening nucleic acid expression libraries; defining the interactions among pathogens, endothelial cells, and neutrophils; and identification of cells which respond specifically to immunomodulatory stimuli by producing cytokines or expressing leukocyte adhesion molecules. Acquisition of the proposed instrument will significantly advance the progress and expand the scope of these studies. Thus, we have identified three principal goals in establishing this research flow cytometry facility on the Harbor-UCLA Medical Center campus: i) to provide an accessible and economical dedicated research flow cytometer for multiple investigators on this campus; ii) to coordinate a flexible network that will enable investigators to perform flow cytometric analyses using fluorochromes and conditions tailored to their specific applications; and iii) to promote technical expertise for investigators with less experience in using modern flow cytometric techniques. These goals will enable investigators to achieve specific aims of ongoing NIH-funded projects, and to generate preliminary data essential for submission of competitive grant applications to the NIH and other agencies. Dedicated space has been allocated by Harbor-UCLA Research and Education Institute to house the instrument and the institution has committed strong support to the development of this essential research facility through coordination of recharge fees, and provisions for maintenance support and overhead costs for its long-term use.