ACAS- WORK STATION FOR ANCHORED CELL ANALYSIS & SORTING

荧光细胞术在过去十年中已经发展成为一种 强大的方法，已经发现了许多应用程序， 细胞生物学，从形态学表征， 细胞结构对化学动力学的测定 一个细胞内的反应。 三大因素同等重要 对这一进展的重要贡献;发展广泛的 各种荧光探针，其中许多是非常具体的 并可用于活细胞;仪器的改进 其中激光的使用带来了显著的提升;以及， 通过软件的进步， 介绍了样品的处理和定量分析方法。 虽然 这些领域的进一步发展是值得欢迎的， 不可避免的是，最先进的技术已经成熟到足以要求 新的实验设计，应阐明生理和 在亚细胞水平上的病理过程。 本提案的目的是确保资金和购买 适当的工具，使一组调查人员 将数字成像荧光显微镜应用于 各自的项目。 这些项目包括研究 脂蛋白亚型在动脉粥样硬化形成和逆转胆固醇中的作用 转运;载脂蛋白B和 细胞内胆固醇沉积;细胞脂质变化受 载脂蛋白C-III;脂蛋白合成的调节， 在造血组织内产生B淋巴细胞; 风湿性疾病中自身抗体的产生; 细胞活化中的膜流动性。 拟议 仪器，“ACAS-470”锚定细胞分析工作站 和分选，代表了显微镜，激光， 微定位和计算机技术。 ACAS-470允许 定量荧光图像分析，荧光分析 光漂白后再分布（FRAP）以及自动 在培养物中选择贴壁依赖性细胞。 这 性能的组合非常适合各种 在所包括的项目内设计的实验方法。 ACAS-470是目前市场上唯一一款 允许荧光分析，荧光重新分布后， 光漂白（FRAP）和贴壁依赖细胞分选 细胞

Fluorescence cytometry has developed over the last decade into a powerful methodology that has found numerous applications in cell biology ranging from morphological characterization of cellular structures to determination of kinetics of chemical reactions within a single cell. Three major factors made equally important contributions to this progress; development of a wide variety of fluorescent probes, many of which are quite specific and can be used in the living cell; improvement of instrumentation in which the use of laser gave a significant boost; and, advancement of software through which a digital image processing and quantitative analysis was introduced. Although further developments in these areas are both welcome and inevitable, the state of the art is sophisticated enough to call for new experimental designs which should elucidate physiological and pathological processes at subcellular levels. The purpose of this proposal is to secure funds and purchase an adequate instrument which would enable a group of investigators to apply digital imaging fluorescence microscopy in their respective projects. These projects include studies of the role of lipoprotein subspecies in atherogenesis and reverse cholesterol transport; structural alterations of apolipoprotein B and cholesterol deposition in cells; cellular lipid changes modulated by apolipoprotein C-III; regulation of lipoprotein synthesis, production of B lymphocytes within hemopoietic tissues; autoantibody generation in rheumatic diseases; and the role of membrane fluidity in cellular activation. The proposed instrument, "ACAS-470" Workstation for Anchored Cell Analysis and Sorting, represents an integration of microscope, laser, micropositioning and computer technologies. ACAS-470 allows quantitative fluorescence image analysis, analysis of fluorescence redistribution after photobleaching (FRAP) as well as automated selection of anchorage dependent cells in culture. This combination of performances is well-suited to the variety of experimental approaches designed within included projects. ACAS-470 is currently the only instrument on the market that allows fluorescence analysis, fluorescence redistribution after photobleaching (FRAP) and cell sorting of anchorage dependent cells.