Zeiss PALM Laser Capture Microdissection System

描述（由申请人提供）：本申请是为购买蔡司PALM微束激光显微解剖和压力弹射（LMPC）系统申请资金。该请求中提出的系统设置是量身定制的，可在大量使用的成像核心设施中具有多种应用，并将充分利用PALM显微解剖系统提供的独特功能。该系统将(1)由一个加热的CO2孵育室封闭，用于长期活细胞成像；(2)将配备蔡司apoome结构照明网格，用于合理快速的高分辨率活细胞成像。(3)该系统还将有两个不同的数字，以满足不同的成像需求，为明视场和低光活细胞成像。这种配置使该系统与常规的显微解剖系统区别开来，因为它非常适合进行组织标本的显微采集，以及亚细胞显微解剖和活细胞显微镜的组合应用。具体来说，该系统是专门配置以满足在这个频繁使用的细胞成像设备中遇到的广泛需求，满足以下应用的需求：(1)厚标本的获取，(2)档案组织标本的微采集，(3)活细胞的采集用于随后的再培养，(4)细胞过程的微切断，(5)活细胞单层的图案损伤，(6)部分核膜的微解剖。我们与卡尔蔡司公司密切合作，在我们的成像设备中挑战了PALM系统，具有非常广泛的要求应用。我们在这里提供了我们建议在系统上执行的每个应用程序的初步数据，从而提供了重要的证据，证明这确实是一个能够提供我们预期和需要的所有工具效益的系统。公共卫生相关性：这种特殊配置的PALM激光显微解剖系统将加强许多基础和转化研究项目。在可容纳300个实验室的核心设施中安装这一多功能系统将确保最大限度地利用其全部优势。

DESCRIPTION (provided by applicant): 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. PUBLIC HEALTH RELEVANCE: 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.