An arsenal of potent assay methods is available for the analyses of genes and the products of their expression. These new tools require highly purified DNA, RNA or protein samples. To most effectively study disease processes, investigators should ideally obtain their samples from a precisely defined organ substructure or a homogeneous population of cells isolated from organs or tissues that are usually quite heterogeneous in cellular makeup. This task may be difficult if the cells of interest constitute a low percentage of the total tissue volume. Laser capture microdissection (LCM) allows the identification of specific sub-structures or cell types and their precise removal from the tissue of origin without damage. LCM offers important advantages over other techniques for cell purification, particularly with respect to precision, speed, simplicity, and archiving capabilities. The acquisition of a Pix Cell lie LCM System at the SIU School of Medicine (SIUSM) will allow our NIH-funded faculty to collect highly defined samples for subsequent genomic analysis. Although our faculty is already experienced at conventional PCR, quantitative real-time PCR (QRT-PCR) offers numerous advantages. Integration of QRT-PCR capability with the Pix Cell lIe LCM system will allow participants to accurately quantify gene expression and to link changes in expression to specific cell populations. As an accessory to the LCM, we are therefore requesting four Smart Cycler II 16-site, add-on blocks. These modules will be used to expand our existing but very limited QRT-PCR capabilities to support a multi-user environment SIUSM and its core Research Imaging Facility will provide the necessary infrastructure and financial support to keep the requested equipment in excellent working order and to promote optimal use by participating faculty. The requested LCM and complementary QRT-PCR instrumentation will allow participating faculty to acquire additional high quality data in support of their current NIH-funded research programs (NHLBI, NINDS, NCRR, NIA, NCI, NICHD, NIDCD). The precise and definitive results afforded by LCM and QRT-PCR are not possible or feasible with existing technology at the SIUSM. The availability of LCM with integrated QRT-PCR as a core facility will greatly strengthen the on-going strong NIH-funded research programs of our faculty.
