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样化用户组，包括一个研究机构和来自约翰霍普金斯大学两所学校的两个部门，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，过时的徕卡TCS-NT，这一直是综合成像中心（IIC）的主要主力。这个最先进的蔡司共聚焦系统将取代并构成一个显著的升级，从我们目前的徕卡系统方面的能力，最重要的是，可靠性。IIC的研究人员主要由美国国立卫生研究院资助，研究大量细胞生物学/生物工程相关问题，包括但不限于染色质结构和基因表达；膜组织与动力学；microtubule-based汽车;膜贩运;生殖细胞发育和迁移；感觉神经元发育；以及内吞作用的机制。在要求的配置中，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，并能够精确调制激光功率以最大限度地减少光毒性和光漂白。此外，我们的研究人员还需要先进的活细胞成像技术，包括FLIP、FRET、FRAP、顺序/多道扫描、GFP的光激活（非笼化）和4D成像。我们的调查人员所要求的这些功能在我们目前的徕卡TCS-NT上都不存在或不足。

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.