ESTABLISHMENT OF UPGRADED FLOW CYTOMETRY SYSTEM CORE

该提案要求支持新设备，以升级 唯一可用于流式细胞仪和细胞分选的设备 北京大学医学院的调查研究 明尼苏达州。现有的工厂只有一个贝顿-迪金森 FACS IV，购买于1980年。有两个主要的理由 拟议的升级：1)现有设施使用率很高， 即使是常规的分析和整理，也不能满足当前的需求； 2)尽管FACS IV已升级为允许双波长 激励和四参数分析，用户都有研究 这需要或将通过最先进的技术来大大增强 方法需要5个或5个以上的检测器进行分析。示例 包括三色和四色免疫荧光分析和分类 免疫荧光的复杂细胞混合物和组合 用荧光探针分析细胞内钙、PH值或细胞 周期分析。为了满足这些需求，目前主要用户有 不得不依赖外部协作和有限的访问权限 其他机构的乐器。该请求由 该设施的主要用户都有目前的NIH项目，包括 流式细胞术。目前资助的研究的主要重点 由这些赠款支持的是关于淋巴细胞分化和 功能：T和NK细胞的功能成熟；分析 正常和白血病B细胞前体细胞的信号转导 正常T细胞；正常人和正常人的分析和辐射敏感性 白血病祖细胞；人类免疫毒素的评价 骨髓移植。拟议的FACStar Plus系统将 满足对最先进的流式细胞仪和 分类。建议的FACScan系统多了一个探测器 比现有的分析仪器，将提供一种方便、 用户友好的仪器，以满足日益增长的需求 免疫荧光分析，并将免费的主要仪器 细胞分类和高级应用程序。

This proposal requests support for new equipment to upgrade the sole facility for flow cytometry and cell sorting available for research investigation in the Medical School of the University of Minnesota. The existing facility has a single Becton-Dickinson FACS IV, purchased in 1980. There are two major justifications for the proposed upgrade: 1) The existing facility is heavily used and cannot meet current demands even for routine analysis and sorting; 2) Although the FACS IV has been upgraded to allow dual wavelength excitation and four parameter analysis, the users all have research that requires or would be greatly enhanced by state-of-the-art methods requiring 5 or more detectors for analysis. Examples include 3- and 4-color immunofluorescent analysis and sorting of complex cell mixtures and combinations of immunofluorescent analysis with fluoroprobes for intracellular calcium, PH or cell cycle analysis. To meet these needs currently the major users have had to rely on outside collaboration and limited access to instruments at other institutions. The request is supported by the major users of the facility all with current NIH Projects involving flow cytometry. The principal focus of currently funded research supported by these grants is on lymphocyte differentiation and function: functional maturation of T and NK cells; analysis of normal and leukemic B cell precursors; signal transduction in normal T cells; analysis and radiosensitivity of normal and leukemic progenitor cells; and evaluation of immunotoxins in human bone marrow transplantation. The proposed FACStar Plus system will meet the acute need for state-of-the-art flow cytometry and sorting. The proposed FACScan system has one more detector for analysis than the existing instrument, will provide an accessible, user-friendly instrument to meet increasing demands for immunofluorescent analysis, and will free the major instrument for cell sorting and advanced applications.