Leica AOBS confocal for Use in Multi-User Facility

描述（由申请人提供）：本提案要求支持采购共聚焦显微镜和远程访问工作站，以添加到部门现有的共享资源中。凯斯西储大学的遗传学教授该系统由安装在全电动Leica DM IRE 2倒置显微镜上的Leica TCS SP2 AOBS无滤波共聚焦、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）它将取代过时且不充分的BioRad MRC 600共聚焦，该共聚焦具有不可靠的步进电机，在图像采集方面的能力有限，并且没有3D重建 和改造软件，以及2）它将更新我们的设施，以便可以进行活细胞成像。 由于目前工厂中的显微镜都没有环境控制或电动部件，因此我们执行活细胞研究和复杂研究任务的能力有限，这些研究任务依赖于高测量精度和大量数据的处理。选择Leica AOBS而不是其他共焦镜是因为它：a）比任何其他系统具有高30-50%的荧光发射灵敏度; B）需要更少的破坏性照明光功率，这对于活细胞应用是一个优势; c）允许在高度散射的样本中进行更深的成像; d）可以使用9条激光线的任何组合; e）允许4个荧光团的同时成像;以及f）具有消除完全重叠的发射光谱的串扰的软件。该工作站将安置在当前的共享设施中，用于检查染色体、活细胞、胚胎和用多种荧光探针标记的生物体。它将补充两个已经满负荷使用的宽视场显微镜工作站。成像主任和咨询委员会将监督系统的管理。大学将提供空间分配，装修，安装，维护和工资的成像主任的支持。本提案中概述的项目取决于共定位，其中最大化重叠发射光谱的分辨率和颜色分离是必不可少的，并且需要测定分子的亚细胞分布和执行3-D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。基于技术考虑和共焦系统之间的比较，我们确定Leica AOBS共焦是满足CWRU教师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 This system will serve two functions: 1) it will replace an outdated and inadequate BioRad MRC600 confocal that has an unreliable stepper motor, limited capabilities in terms of image acquisition, and no 3-D reconstruction &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.