PURCHASE OF A FLOW CYTOMETER/CELL SORTER

该提案要求为购买一台流式细胞仪提供资金。 (具有分类能力)，用于该部的几名调查员 佛罗里达大学比较与实验病理学。这项研究 这些研究人员集中在各种生物制剂的功能上 例如病毒、毒素、激素和免疫调节剂以及 与细胞表面各自的受体相互作用，以及 细胞表面抗原/标记物的鉴定和定量 细胞的表面。我们目前的努力因缺乏准入而受到阻碍 到快速、灵敏的技术来研究单细胞的这种相互作用 和/或分子水平。尽管许多特工都是 被研究的物质只以极少量的形式存在于体内，我们努力 通过研究他们的行动模式得到了相当大的推动 几家生物技术公司使用DNA生产的材料的可用性 重组技术。随时准备使用流式细胞仪将使我们能够 在单细胞水平上研究这些试剂的功能活性 具有标准采样无法达到的时间分辨率 使用总细胞群体的技术。 我们计划的未来实验也取决于生产 单抗作为各种细胞的特异和独特的标志物 以及它们不同的受体。在这种标记的帮助下，相互作用 对不同细胞和不同细胞功能的调节 分析过了。尽管克隆抗体的生产已经成为一种 我科常规技术，分离、鉴定和鉴定 维持产生特异性抗体的杂交克隆是一段时间 消费过程。流式细胞仪极大地减少了所需的时间 以寻找和维持产生的单抗与 所需的特异性，因为它的排序功能缩短了选择范围 和克隆程序。

This proposal requests funding for the acquisition of a Flow Cytometer (with sorting capabilities) for several investigators in the Department of Comparative & Experimental Pathology, University of Florida. The research of these investigators centers on the function of various biological agents such as viruses, toxins, hormones, and immune regulators and the interaction with their respective receptors on cell surfaces, as well as identification and quantification of antigens/markers expressed on the surfaces of cells. Our current efforts are handicapped by lack of access to fast, sensitive techniques to study such interactions at the single cell and/or the molecular level. Although many of the agents that are being studied are only present in the body in minute quantities, our efforts to study their mode of action have received a considerable boost through the availability of material produced by several biotechnology firms using DNA recombinant techniques. Ready access to a flow cytometer would allow us to study the functional activity of such reagents at the single cell level with a time resolution that is impossible to achieve with standard sampling techniques using total cell populations. Future experiments we have planned are also dependent on the production of monoclonal antibodies as specific and unique markers for the various cells and their different receptors. With the aid of such markers the interplay and regulation of different cells and different cellular functions can be analyzed. Although the production of monoclonal antibodies has become a routine technique in our department, the isolation, characterization and maintenance of hybrid clones producing specific antibodies is a time consuming process. A flow cytometer drastically reduces the time required to search for and maintain the production of monoclonal antibodies with the desired specificity because its sorting capabilities shortens the selection and cloning procedures.