Laser Capture Microdissection (LCM) System

描述(由申请人提供)：大量有效的分析方法可用于分析基因及其表达产物。这些新工具需要高度纯化的DNA、RNA或蛋白质样本。为了最有效地研究疾病过程，理想情况下，研究人员应该从精确定义的器官亚结构或从器官或组织中分离出来的同质细胞群中获取样本，这些器官或组织的细胞构成通常是相当不同的。如果感兴趣的细胞占总组织体积的百分比很低，这项任务可能会很困难。 激光捕获显微切割(LCM)可以识别特定的亚结构或细胞类型，并将其从来源组织中精确移除，而不会造成损害。与其他细胞纯化技术相比，LCM具有重要的优势，特别是在精密度、速度、简单性和归档能力方面。在SIU医学院(SIUSM)收购Pix Cell Lie LCM系统将使我们由NIH资助的教职员工能够收集高度明确的样本，用于后续的基因组分析。虽然我们的教员在常规的聚合酶链式反应方面已经很有经验，但定量实时聚合酶链式反应(QRT-PCR)提供了许多优点。QRT-PCR能力与Pix Cell Lie LCM系统的集成将使参与者能够准确量化基因表达，并将表达变化与特定细胞群体联系起来。因此，作为LCM的附件，我们需要四个16站点的Smart Cycler II附加模块。这些模块将用于扩展我们现有但非常有限的QRT-PCR能力，以支持多用户环境 SIUSM及其核心研究成像设施将提供必要的基础设施和财政支持，以保持所要求的设备处于良好的工作状态，并促进参与的教员最佳使用。所要求的LCM和补充的QRT-PCR仪器将允许参与的教师获得额外的高质量数据，以支持他们目前由NIH资助的研究项目(NHLBI、NINDS、NCRR、NIA、NCI、NICHD、NIDCD)。在SIUSM的现有技术下，LCM和QRT-PCR提供的准确和确定的结果是不可能或不可行的。LCM与整合的QRT-PCR作为核心设施的可用性将极大地加强我们教员正在进行的强大的NIH资助的研究项目。

DESCRIPTION (provided by applicant): 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.