| Description |
The Harbor-UCLA Medical Center has a state-of-the art morphology core capable of performing morphological (both at cellular and subcellular levels), biochemical, cell and molecular biological studies. This newly established core will provide consultation and training services for immunohistochemistry and confocal microscopy for localization as well as subcellular distribution of proteins in tissues. This core has a Zeiss microscope equipped with Axiocam color digital camera, Axiovision, Image Pro-Plus software and an advanced computer workstation with graphic accelerators and high storage capacity for image processing. It also houses a Leica TCS SP Upright Confocal Microscope which has three lasers, an argon, a krypton, and a helium-neon which are connected to the microscope by a fiber optic and produce laser lines at 476 (near blue) 488 (blue), 568 (yellow) and 633(far red) nm. The system is equipped with three reflectance/fluorescence photomultiplier/ detectors and one for transmitted light (bright field, phase, and darkfield, etc.). In addition the facility houses a PALM (stands for positioning and ablation with laser microbeams) microdissection system, which uses Laser Pressure Catapulting technology to dissect selected specimens from various sources. This system, which enable fast, easy and reliable non-contact laser micromanipulation and microdissection, consists of a 337 nm pulsed nitrogen laser, a wave length which does not interfere biological function, RoboStage, CCD color video camera, fluorescence module for visualization of GFP and 488 nm fluorescent markers, manipulator, and a Zeiss Axiovert 200 microscope. The PALM-based molecular analyses of selected tissue and cell samples provide a better insight into the molecular components of certain disease processes or cellular pathways. Coupled with acquisition of the laser capture instrumentation, the availability of real-time PCR should allow the quantification of low abundance mRNAs in samples derived from either single cells or from small cell clusters. The availability and introduction of this system together with the expertise on histomorphology and cell biology provided by Dr. Sinha-Hikim will allow the molecular analyses of complex tissues and expand the GCRC core facilities to recruit larger numbers of potential NIH-funded clinical as well as serve basic investigators. The PALM microdissection system represents the up-stream tissue processing required before molecular analyses. The acquisition of the PALM instrument has allowed investigators from diverse disciplines and sites to harvest homogeneous populations of cells from tissues of interest before molecular analyses using the other resources and equipment of the proposed Core.
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