RESEARCH FLOW CYTOMETER

与传统的显微荧光测定法或放射自显影法相比，流式细胞术具有显著的优势。首先，流式细胞术允许快速分析单个细胞或细胞群内的分布。 通过这种方式，可以分析来自给定群体的大量细胞，甚至可以检测到最罕见的亚群。 具有增加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。 染色技术和更好地保持细胞活力和膜完整性的条件的进步使得能够通过流式细胞术详细分析细胞周期和抗原表达。 流式细胞术与细胞分选的互补提供了额外的优点，即一旦通过荧光鉴定细胞具有不寻常或所需的特征，就可以物理分离细胞以进行进一步研究。 这项建议是基于迫切需要利用流式细胞术的这些优势，以实现正在进行的NIH资助的研究的具体目标。 研究者将使用流式细胞术技术来实现这些目标，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粘附分子来识别对免疫调节刺激有特异性反应的细胞。 获得拟议的仪器将大大推进这些研究的进展并扩大其范围。 因此，我们已经确定了在Harbor-UCLA医学中心校园内建立这种研究流式细胞仪设施的三个主要目标：i）为该校园内的多个研究者提供一种可访问的和经济的专用研究流式细胞仪; ii）协调一个灵活的网络，使研究者能够使用荧光染料和针对其特定应用的条件进行流式细胞仪分析;及iii）向在使用现代流式细胞技术方面经验较少的研究人员推广技术专长。 这些目标将使研究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资助申请所必需的初步数据。 Harbor-UCLA研究和教育研究所分配了专用空间来容纳该仪器，该机构通过协调充电费，以及为其长期使用提供维护支持和间接费用，致力于大力支持这一重要研究设施的发展。

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.