Purchase of Leica TCS SP2 Confocal Microscope System

此共享仪器赠款申请用于购买徕卡TCS SP2激光扫描共聚焦显微镜系统。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核心资源。恩格勒实验室成立于1981年，用于流式细胞术，自1993年以来一直为达特茅斯提供成像和流式细胞仪仪器。新的共焦仪器将取代过时的共焦系统，使用户能够获得他们所需的稳定、灵活和最新的成像能力。对新的共焦系统的需求是基于几个紧迫的需求。现有的共焦系统不具有激光稳定性或激发和发射灵活性，这是由不同和大量的用户进行多色定量工作所需的。此外，存在对活细胞研究的明显需求，而现有的直立显微镜支架缺乏温度控制无法满足这一需求。最后，一个新的系统将具有获取12位强度分辨率的图像的能力，将为FRAP和反向FRAP分析提供感兴趣的区域，并将为四种颜色的荧光采集提供4个PMT-所有这些都是我们目前的系统所缺乏的，现在许多用户需要这些。徕卡TCS SP2系统将满足核心设施的当前用户，并将提供灵活性，以适应未来用户的需求。恩格勒特细胞分析实验室在一名主任、一名联席主任和一个咨询委员会的控制下运作，并在流程和成像主管的技术支持下运行。它的财政支持来自NCI对诺里斯棉花癌症中心的核心拨款，对用户的每小时收费，以及达特茅斯医学院的机构支持。目前，有33名NIH赠款持有人依赖恩格勒特核心实验室的共焦系统；他们作为私人投资机构持有50笔NIH赠款，每年的直接成本为9918,884美元。已经确定了一个“主要用户群”。这11个主要用户预计占共焦系统总使用量的84%。作为私人投资机构，他们拥有20笔NIH拨款，每年的直接成本为5,161,571美元。

This Shared Instrumentation Grant application is for the purchase of a Leica TCS SP2 laser scanning confocal microscope system. The instrument will be integrated into the Englert Cell Analysis Laboratory, which is a Shared Core Resource of the Norris Cotton Cancer Center at Dartmouth Medical School. The Englert Laboratory was establish3d in 1981 for flow cytometry and has been providing Dartmouth with imaging as well as flow cytometric instrumentation since 1993. The new confocal instrument will replace an outdated confocal system, giving users access to the stable, flexible, and up-to-date imaging capabilities that they require. The requirement for a new confocal system is based on several urgent needs. The existing confocal system does not have the laser stability or excitation and emission flexibility required for multicolor quantitative work by a varied and large group of users. In addition, there is an expressed need for live cell studies that cannot be met by the existing upright microscope stand lacking temperature control. Finally, a new system will have the ability to acquire images with 12 bit intensity resolution, will provide regions of interest for FRAP and inverse FRAP analysis, and will provide 4 PMTs for four color fluorescence acquisition-all of which are lacking on our current system and are required now by many users. A Leica TCS SP2 system will satisfy current users of the Core Facility and will provide flexibility for adaptation to the needs of users in the future. 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 an imaging supervisor. It is supported financially from the NCI core grant to the Norris Cotton Cancer Center, by hourly charges to users, and by institutional support from Dartmouth Medical School. At the present time, there are 33 NIH grant holders dependent upon the confocal system in the Englert Core Laboratory; they hold, as PIs, 50 NIH grants, with $9,918,884 in annual direct costs. A "major user group" has been identified. These eleven major users project 84% of the total use of the confocal system. They hold, as PIs, 20 NIH grants with $5,161, 571 in annual direct costs.