Andor Revolution XD Live Cell Imaging System with Photomanipulation Laser

描述(申请人提供)：本申请是为了申请资金购买Andor Revsion XD活细胞成像系统，该系统配备旋转圆盘共焦显微镜和Frappa光操纵激光模块，以执行关键的光消融、光激活、光转换。本申请中建议的系统设置是为在具有多种应用的高使用量的成像核心设施中高度通用而量身定做的，并将充分利用Andor旋转磁盘共焦系统所提供的独特功能。该系统将配备(1) FRAPPA激光模块对许多光操纵实验至关重要；以及(2) 将安装在尼康完美聚焦钛倒置显微镜上，在长期成像过程中进行严格的焦面补偿。具体地说，该系统的配置是为了满足大量使用的细胞成像设备中遇到的广泛需求，同时满足以下应用的需求：(I)以每秒5帧速率快速获取高质量图像，(Ii)与成像相关的最小光漂白，(Iii)用于非常长期的活细胞成像的稳定阶段，(Iv)活细胞台顶孵化室(V)用于光调节实验的外部激光器，例如FRAP、Flip、FRET、光转换和光激活。我们在这里提供了我们建议在该系统上执行的应用程序的初步数据，从而提供了重要的证据，表明这确实是一个将提供我们预期和需要的所有工具好处的系统。

DESCRIPTION (provided by applicant): This application is to request funds to purchase the Andor Revolution XD live cell imaging system equipped with spinning disk confocal microscope and the FRAPPA photomanipulation laser module to perform the crucial photoablation, photoactivation, photoconversion. The system set-up proposed in this request is tailor-designed to be highly versatile in a heavily used imaging core facility with a multitude of applications and will take full advantage of the unique features afforded by the Andor spinning disk confocal system. The system will be (1) equipped with the FRAPPA laser module critical for numerous photomanipulation experiments; and (2) will be fitted on Nikon PerfectFocus Ti inverted microscope for rigorous focal plane compensation during long-term imaging. Specifically, this system is configured to cater to the wide-ranging need encountered in the heavily used Cell Imaging Facility by simultaneously meeting the demand of the following applications: (i) fast acquisition of high quality image at > 5 images per second frame rate, (ii) minimal imaging-related photobleaching, (iii) stable stage for very long term live cell imaging, (iv) live cell stage-top incubation chamber (v) external lasers for photomanipulation experiments such as FRAP, FLIP, FRET, photoconversion and photoactivation. We present here preliminary data for the applications we propose to perform on the system, thus provide important evidence that this is indeed the system that will deliver all the instrumental benefits we anticipate and need.