Leica AOBS confocal for Use in Multi-User Facility

说明(由申请人提供)：本建议书请求支持购置一台共焦显微镜和一个远程访问工作站，以便将其添加到部门现有的共享资源中。凯斯西储大学遗传学教授。该系统由安装在全电动徕卡DM IRE2倒置显微镜上的徕卡TCS SP2 AOBS无滤光镜共焦、环境室、无振动工作站、计算机和软件组成。该系统将提供两个功能：1)它将取代过时和不足的BioRad MRC600共焦，它有一个不可靠的步进电机，在图像采集方面能力有限，并且没有3D重建 和重塑软件，以及2)它将更新我们的设施，以便可以进行活细胞成像。 由于目前设施中的显微镜都没有环境控制或电动部件，我们执行活细胞研究和复杂研究任务的能力受到限制，这些研究任务依赖于高测量精度和处理大量数据。之所以选择徕卡AOBS，是因为它比其他任何系统对荧光发射的灵敏度高30-50%；b)需要的破坏性照明光功率小得多，这是活细胞应用的优势；c)允许在高度分散的样品中进行更深层次的成像；d)可以使用9条激光线路的任意组合；e)允许同时对4个荧光团成像；f)具有消除完全重叠发射光谱串扰的软件。该工作站将被安置在目前共享的设施中，用于检查染色体、活细胞、胚胎和用多个荧光探针标记的生物体。它将补充已经满负荷使用的两个广域显微镜工作站。成像主管和咨询委员会将监督该系统的管理。该大学将为成像主任提供空间分配、翻新、安装、维护和工资方面的支持。本提案中概述的项目依赖于共定位，在这些定位中，最大限度地提高重叠发射光谱的分辨率和颜色分离是至关重要的，并需要分析分子的亚细胞分布和执行3D重建的能力。这些要求需要具有最高分辨率、灵敏度和灵活性的共焦显微镜。基于对共焦系统的技术考虑和比较，我们确定徕卡AOBS共焦系统是满足CWRU教职员工成像需求的最佳选择。

DESCRIPTION (provided by applicant): This proposal requests support for the acquisition of a confocal microscope and a remote access workstation to be added to an existing shared resource in the Dept. of Genetics at Case Western Reserve University. The system consists of a Leica TCS SP2 AOBS filter-free confocal mounted on a fully-motorized Leica DM IRE2 inverted microscope, environmental chambers, a vibration-free workstation, computers and software. This system will serve two functions: 1) it will replace an outdated and inadequate BioRad MRC600 confocal that has an unreliable stepper motor, limited capabilities in terms of image acquisition, and no 3-D reconstruction & remodeling software, and 2) it will update our facilities so that live-cell imaging can be performed. Since none of the microscopes in the current facility have environmental controls or motorized parts we are limited in our ability to perform live cell studies and complex research tasks that rely on high measuring accuracy and processing of large amounts of data. The Leica AOBS was chosen over other confocals since it: a) has 30-50% higher sensitivity for fluorescence emission than any other system; b) needs far less damaging illumination light power which is an advantage for live cell applications; c) allows deeper imaging in highly scattered specimens; d) can use any combination of 9 laser lines; e) allows simultaneous imaging of 4 fluorophores; and f) has software that eliminates crosstalk of completely overlapping emission spectra. The workstation will be housed in the current shared facility for examination of chromosomes, living cells, embryos, and organisms labeled with multiple fluorescent probes. It will complement two wide-field microscope workstations already utilized at capacity. The imaging director and advisory committee will oversee management of the system. The University will provide support for space allocation, renovation, installation, maintenance, and salary for the imaging director. The projects outlined in this proposal depend upon co- Iocalizations where it is essential to maximize resolution and color separation of overlapping emission spectra, and require the ability to assay the subcellular distribution of molecules and perform 3-D reconstruction. These requirements necessitate a confocal microscope with the highest resolution, sensitivity, and flexibility. Based on technical considerations and comparisons between confocal systems, we have determined that the Leica AOBS confocal is the best choice to meet the imaging needs of the faculty at CWRU.