Zeiss LSM 510 META Confocal Microscope

描述（由申请人提供）：一个由8名研究人员组成的多元化用户组，包括约翰霍普金斯大学两所学校的一个研究所和两个部门，要求资金购买蔡司LSM 510 Meta共聚焦显微镜，以取代大量使用的过时Leica TCS-NT，该显微镜一直是集成成像中心（IIC）的主要主力。这种最先进的蔡司共焦系统将取代我们现有的徕卡系统，并在功能和最重要的可靠性方面进行重大升级。IIC研究人员主要通过NIH资助，致力于研究许多细胞生物学/生物工程相关问题，包括但不限于染色质结构和基因表达;膜组织和动力学;基于微管的马达;膜运输;生殖细胞发育和迁移;感觉神经元发育;和内吞作用机制。在所要求的配置中，LSM 510-Meta将为我们的研究人员提供同时精确成像多个荧光团的能力，以及精确调制激光功率以最大限度地减少光毒性和光漂白的能力。此外，我们的研究人员还需要先进的活细胞成像技术，如FLIP、FRET、FRAP、顺序/多轨道扫描、GFP的光激活（解除锁定）和4D成像。我们的研究人员所需的这些功能在我们目前的Leica TCS-NT上都不存在或不足。 在本次提交中，更重要的是我们目前的徕卡系统始终不可靠;以及徕卡服务无法将我们的系统保持在合格的工作状态。虽然自1998年收购以来一直根据服务合同进行维护，但其使用寿命的25%已无法使用。此外，即使在可操作的情况下，由于持续的和无法解决的激光/针孔/物镜问题，系统在近40%的时间内的性能也低于最佳。因此，考虑到共聚焦显微镜对IIC研究人员的核心重要性，以及对可靠性和性能限制的担忧不断升级，我们主动租用了LSM 510 Meta，以便1）确保我们的用户可靠地使用共聚焦显微镜，从而支持NIH资助的研究;以及，2）在预计这项拨款申请时，随时准备好评估我们研究的性能和适用性的工具。重要的是，租赁该系统并不意味着必须购买;而是允许LSM 510-Meta与巴尔的摩地区的其他竞争仪器（包括Leica TCS-SP2和Perkin-EImer Ultraview）“并排”进行全面评估。在我们的用户组的评估中，LSM-510 Meta在多功能性、能力和对拟议研究的适用性方面显然是最好的。

DESCRIPTION (provided by applicant): A diverse user-group of 8 investigators, encompassing one research institute and two departments from two Schools at The Johns Hopkins University, request funds to purchase a Zeiss LSM 510 Meta confocal microscope to replace a heavily used, antiquated Leica TCS-NT, which has been the primary workhorse of the integrated Imaging Center (IIC). This state-of-the-art Zeiss confocal system will replace and constitute a significant upgrade from our present Leica system in terms of capabilities and, most importantly, reliability. IIC investigators, funded primarily through the NIH, work on a host of cell biological/bioengineering related questions including, though not limited to, chromatin structure and gene expression; membrane organization and dynamics; microtubule-based motors; membrane trafficking; germ cell development and migration; sensory neuron development; and mechanisms of endocytosis. In the requested configuration, the LSM 510-META will afford our investigators the capability to precisely image multiple fluorophores simultaneously, and the ability to accurately modulate laser power to minimize phototoxicity and photobleaching. Additionally, our investigators variously require the advanced live cell imaging techniques of FLIP, FRET, FRAP, sequential/multi-track scanning, photo-activation (un-caging) of GFP, and 4D imaging. These capabilities required by our investigators are all absent or deficient on our current Leica TCS-NT. Of greater import in this submission has been the consistent unreliability of our current Leica system; and the inability of Leica-Service to maintain our system in competent working condition. Though maintained under service contract since the instrument was acquired in 1998, it has been non-functional 25% of its useful life. Moreover, even when operable, the system performed less than optimally nearly 40% of the time due to ongoing and unresolvable laser/pinhole/ objective problems. Consequently, given the central importance of confocal microscopy to IIC investigators, and escalating concerns pertaining to reliability and performance limitations we proactively undertook the lease/rental of an LSM 510 META in order 1) to ensure reliable access to confocal microscopy for our users thus supporting NIH funded research; and, 2) in anticipation of this grant application, have the instrument readily available for evaluation as to performance and suitability in our research. Importantly, renting the system did not obligate its purchase; but rather allowed for thorough evaluation of the LSM 510-META 'side-by-side' against other competitive instruments available in the Baltimore area, including the Leica TCS-SP2 and the Perkin-EImer Ultraview. In the evaluation of our user-group, the LSM-510 Meta is clearly the best in terms of versatility, capability, and suitability for the proposed research.