This application is to request funds to purchase the Zeiss PALM MicroBeam Laser Microdissection and Pressure Catapulting (LMPC) System. 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 PALM microdissection system. The system will be (1) enclosed by a heated CO2 incubation chamber for long term live cell imaging and (2) will be equipped with Zeiss Apotome structured illumination grid for reasonably fast high resolution live cell imaging. (3) The system will also have two different digital to cater for the distinct imaging need for brightfield and low-light live cell imaging. This configuration thus sets this system apart from a regular microdissection system as it is perfectly suited to perform micro-harvesting of tissue specimens as well as the combinatorial application of subcellular microdissection and live cell microscopy. Specifically, this system is specifically configured to cater to the wide-ranging need encountered in this heavily used Cell Imaging Facility by meeting the demands of the following applications: (1) procurement of thick specimen, (2) micro-harvesting of archival tissue specimen, (3) live cell harvesting for subsequent recultivation, (4) micro-severing of cellular processes, (5) patterned wounding of live cell monolayer, and (6) microdissection of portion of nuclear membrane. Working closely with Carl Zeiss Inc., we have challenged the PALM system in our imaging facility with this remarkably wide spectrum of demanding application. We present here preliminary data for each and every application 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.
This specially configured PALM Laser Microdissection System will enhance many basic and translational research projects. The installation of this versatile system in a core facility catering for ~300 labs will ensure that the maximum utilization of its full advantage.
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