SHARED FLOW CYTOMETRY FACILITY (FACSTAR PLUS)

这项提案是为了资助FACStarplus流式细胞仪/细胞分选机 更换现在由纽约大学流式细胞仪设备操作的Ortho 50H。 Ortho 50H虽然在当时非常先进，但却是一款笨拙的 用作分拣器的仪器。除了这种尴尬之外，它还 限制了它可以分析的参数的数量以及 其10年历史的计算机运行的速度。它也正在成为 一个“孤儿”乐器，没有得到制造商的支持。纽约大学 流式细胞仪设施是一个核心实验室，为 整个医疗中心。它的设计目的是提供“最先进的” 为支持700多万美元同行审查提供资金的设施 研究。 本建议书中要求的FACStarplus提供以下能力 用Ar激光进行3色分析，用4色分析 氩激光和染料激光的结合。三个荧光信号可以 用一台激光进行分析(和分类)。其中一个激光器有紫外线 使Hoechst染料可用于DNA检测的能力 活细胞和排序可以基于位置在 细胞周期。还包括染色体分选和流式核型分析。 在该仪器的能力范围内，细胞内钙、pH和 膜粘度的测定也可以使用紫外线 激动人心。去除染料泵后，该激光器可用于 荧光素和藻红蛋白，而另一种激光用于 紫外线。15usec排序周期允许以较高的速度进行排序 接近5000个细胞/秒。这比Ortho快三到四倍。 实际上，这意味着一种设计为产生5x10(7) 用于生化分析的细胞或从某一来源生产的cDNA文库 其中所需细胞占种群的20%可以是 在12小时内完成，而不是两天。

This proposal is for fund to a FACStarplus flow cytometer/cell sorter to replace the Ortho 50H now operated by the NYU Flow Cytometry Facility. The Ortho 50H, while remarkably advanced for its time, is an awkward instrument to use as a sorter. In addition to this awkwardness, it is limited in both the number of paramaeters that it can analyze and by the speed at which its 10 year old computers operate. It is also becoming an "orphan" instrument, poorly supported by its manufacturer. The NYU Flow Cytometry Facility is a core laboratory and provides service to the entire medical center. It is designed to provide "state of the art" facilities to support more than $7 million in peer review funded research. The FACStarplus requested in this proposal provides the capacity to perform 3 color analysis with the argon laser and 4 color analysis with the combination of argon and dye lasers. Three fluorescence signals can be analyzed (and sorted) with a single laser. One of the lasers has UV capability so that Hoechst dyes can be used for DNA determinations on viable cells and sorting can be performed on the basis of position in the cell cycle. Chromosomal sorting and flow karyotyping are also within the capabilities of this instrument Intracellular Calcium, pH and membrane viscosity determination can also be performed using UV excitation. With the dye pump removed, this laser can be used for fluorescein and phycoerythrin while the other argon laser is used for UV. The 15usec sort cycle permits sorting with high recoveries at rates close to 5000 cells/sec. This is three to four times faster the Ortho. In practical terms this means that a sort designed to produce 5 x 10(7) cells for biochemical analysis or cDNA library production from a source in which the desired cells make up 20% of the population can be performed in 12 hours not two days.