Multi-Parameter Flow Cytometer with High-Speed Sorting

描述（由申请人提供）：本共享仪器资助申请用于购买一台Becton Dickinson FACSAria高速分选流式细胞仪。该仪器将被整合到Englert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需要的最新的分类和分析功能。对新的流式细胞仪的需求是基于达特茅斯目前可用的细胞仪无法满足的几个需求。主要需要的是通过四个以上的荧光参数来定义细胞的能力。FACSAria将有三个激光器，允许用户在光谱上激发一系列荧光色，并对13种荧光参数中的任何一种（或全部）进行细胞分析和分选。此外，FACSAria将允许用户通过一次分选四个细胞群来节省时间和细胞样本；将允许交叉光束补偿，以方便使用重叠的荧光染料；将方便定义具有非矩形排序门的单元格；并且将在高细胞流速下提供更有效的分选。Englert细胞分析实验室在一名主任、一名联合主任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。运行费用由NCI对诺里斯棉花癌症中心的核心拨款补贴、按小时向使用者收费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购买FACSAria的费用提供额外的机构支持。目前，有47名NIH资助持有人依赖于Englert核心实验室的流式细胞仪；作为私人律师，他们拥有88项NIH拨款。已确定“主要用户组”。这9个主要用户占FACSAria细胞仪总使用量的78%。他们以pi的身份持有NIH的拨款。

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.