This proposal requests support for the acquisition of an Arcturus AutoPix Laser Capture Microdissection Instrument (LCM) to be added to an existing shared histology/imaging resource in the Visual Sciences Research Center (VSRC) at Case Western Reserve University (CASE). A Users Group of 14 NIH-funded investigators utilize gene/protein expression-profiling technology, but are severely limited in their ability to select specific cell populations for high level genomic/proteomic analyses. The Arcturus AutoPix, the latest generation LCM, is a self-contained system for acquisition of specific tissues or cells from frozen sections for genomic and/or proteomic studies. The AutoPix was chosen over other commercial systems since it is the only LCM with (a) cell/tissue pattern recognition to automatically select cells as trained by the user, (b) multi-slide capacity to increase efficiency, (c) ability to use material without any special mounting or preparation requirements, and (d) an enclosed, protected environment to minimize mRNA/protein contamination. Moreover, Arcturus provides full technical and reagent support for isolation and linear amplification of mRNA for use in DNA microarray studies. The University will provide dedicated space for the LCM and the VSRC will provide salary for both the Resource Manager and Research Assistant III. The PI and Advisory Committee will oversee management of the system. The projects outlined in this proposal require a high-throughput system for the collection of specific cell types for analyses of gene and protein expression using DNA microarray, quantitative PCR, and mass spectroscopy. These requirements necessitate an instrument with the ability to rapidly identify and capture desired cell types in numbers to yield nucleic acid/protein sufficient in quality and quantity for subsequent analyses. This represents a partnership between the CASE VSRC and Cancer Center, and users will interact with excellent existing Center resources in histology, imaging, gene expression analysis, and bioinformatics, collectively functioning as an integrated, University-wide resource. Based upon technical specifications and instrument comparisons, we determined that the AutoPix is the best choice to meet the needs of CASE faculty.
