The objectives of the Microdissection and Cell Acquisition Module are to provide the equipment, technical service, training and expertise for the acquisition of individual cells or groups of cells for use in gene profiling and to provide investigators with optical imaging, digital image processing and image analysis capabilities. The specific goals of this module are to: (i) assist investigators in the preparation and sectioning of ocular tissues for laser capture microdissection;(ii) isolate and capture single cells or groups of cells from desired areas of the eye using the P.A.L.M. MicroBeam microdissection system;(iii) isolate RNA from extracted cells;(iv) train investigators in RNA amplification techniques for subsequent use in gene profiling;and (v) assist in state-of-the-art light and fluorescent microscopic image capture, and analysis. The module will fulfill the fundamental needs of our investigators to precisely isolate either single cells or clusters of cells from defined regions of ocular tissues (e.g., slow-cycling limbal epithelial basal cells versus rapidly cycling corneal epithelial basal cells;pigmented versus non-pigmented ciliary epithelia;corneal epithelial basal and wing cells before and after injury) that are either in a resting or perturbed (diseased) state. Isolated cells will be used for gene expression profiling, or cell culture experiments utilizing the Tissue and Cell Culture Module.
