Multi-Parameter Flow Cytometer with High-Speed Sorting

描述(申请人提供)：这份共享仪器助学金申请是为了资助购买Becton Dickinson FACSAria高速分选流式细胞仪。该仪器将被整合到恩格勒特细胞分析实验室，该实验室是达特茅斯医学院诺里斯棉花癌症中心的共享资源。新的流式细胞仪将为用户提供他们所需的最新分选和分析能力。对新的流式细胞仪的要求是基于达特茅斯目前提供的细胞仪无法满足的几个需求。主要的需求是能够通过四个以上的荧光参数来定义细胞。FACSAria将有三个激光器，允许用户激发光谱上的一系列荧光染料，并根据13个荧光参数中的任何一个(或全部)对细胞进行分析和分类。此外，FACSAria将允许用户一次分选四组细胞，从而节省时间和细胞样本；将允许交叉光束补偿，以便于使用重叠的荧光染料；将促进具有非矩形分选门的细胞的定义；将在高细胞流速下提供更有效的分选。恩格勒特细胞分析实验室在一名董事、一名联席董事和一个咨询委员会的控制下运作，并在一名流动和成像主管的技术支持下运作。运营成本由NCI对诺里斯棉花癌症中心的核心拨款补贴、向用户收取的每小时费用以及达特茅斯医学院的机构支持来支付。达特茅斯医学院将为FACSAria的购买费用提供额外的机构支持。目前，有47名NIH赠款持有者依赖恩格勒特核心实验室的流式细胞仪；他们持有88笔NIH赠款。已经确定了一个“主要用户群”。这九个主要用户预计占FACSAria细胞仪总使用量的78%。作为私人投资机构，他们持有NIH拨款。

DESCRIPTION (provided by applicant): This Shared Instrumentation Grant application is for funding for the purchase of a Becton Dickinson FACSAria high-speed sorting flow cytometer. The instrument will be integrated into the Englert Cell Analysis Laboratory, which is a shared resource of the Norris Cotton Cancer Center at Dartmouth Medical School. The new flow cytometer will provide users with the up-to-date sorting and analysis capabilities that they require. The requirement for a new flow cytometer is based on several needs that are not met by the cytometers currently available at Dartmouth. The primary need is for the ability to define cells by more than four fluorescence parameters. The FACSAria will have three lasers and will permit users to excite a range of fluorochromes across the spectrum and to analyze and sort cells on any (or all) of thirteen fluorescence parameters. In addition, the FACSAria will allow users to save time and cell sample by the sorting of four populations of cells at one time; will allow cross-beam compensation to facilitate the use of overlapping fluorochromes; will facilitate the definition of cells with non-rectangular sort gates; and will provide for more efficient sorting at high cell flow rates. The Englert Cell Analysis Laboratory operates under the control of a Director, a Co-Director, and an Advisory Board, and with the technical support of a flow and imaging supervisor. Running costs are met by a subsidy from the NCI core grant to the Norris Cotton Cancer Center, by hourly charges to users, and by institutional support from Dartmouth Medical School. Additional institutional support will be provided by Dartmouth Medical School toward the purchase cost of the FACSAria. At the present time, there are 47 NIH grant holders dependent upon the flow cytometers in the Englert Core Laboratory; they hold, as PIs, 88 NIH grants. A "major user group" has been identified. These nine major users project 78% of the total use of the FACSAria cytometer. They hold, as PIs, NIH grants.