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.
