Molecular/Morphology Core

分子和形态核心设施为五个单独的项目提供最先进的专业知识，涉及各种分子和结构相关技术。分子组分在组织和分散分离的Cajal间质细胞、肠神经细胞和平滑肌细胞的RT-PCR领域提供专业知识。分子组成部分继续为分子生物学研究提供基因组克隆、cDNA和培养的日常维护。核心拥有自动DNA测序仪(ABI 310和ABI 7700)，以对新克隆、扩增产品和转基因样品进行DNA序列分析。这个 分子组件为所有适用的项目和感兴趣的动物提供常规的转基因动物基因分型，设计和测试RT-PCR的引物，并为拟议的实验构建载体。核心的这个成分也为表达各种项目特异性cDNA的哺乳动物细胞系提供支持。核心的形态部分提供了各种成像技术方面的专业知识，以支持单个项目的形态需求。这些技术涵盖常规光学显微镜、相差显微镜、荧光显微镜、激光扫描共聚焦显微镜(固定组织的实时和标准三维重建)、数字成像、透射式电子显微镜、原位杂交、免疫组织化学和免疫电子显微镜(免疫金和二氨基联苯胺反应)等领域。该核心最近开发的其他领域包括使用Fluo-4对ICC-My网络进行钙成像，以及实时共聚焦显微镜。该核心还在胃肠道内兴奋性的时空映射领域提供支持。形态核心还将能够在不久的将来为对分析血浆中蛋白质相互作用感兴趣的项目提供FRET分析 膜或细胞质。核心输出的质量可以通过发表在胃肠病学和美国生理学杂志上的几篇论文封面上的图像来评估。

The Molecular and Morphology core facility provides state of the art expertise to the five individual projects on a variety of molecular and structurally related technologies. The Molecular component provides expertise in the areas of RT-PCR on tissues and on dispersed isolated interstitial cells of Cajal, enteric neurons and smooth muscle cells. The molecular component continues to provide day to day maintenance of genomic clones, cDNAs and cultures for molecular biological investigations. The core has automated DNA sequencers (ABI 310, and an ABI 7700) to perform DNA sequence analysis on novel clones, amplification products and transgenic samples. The molecular component provides routine genotyping of transgenic animals for all applicable projects and animals of interest, designs and tests primers for RT-PCR and constructs vectors for proposed experiments. This component of the core also provides support with mammalian cell lines expressing various project specific cDNAs. The morphology component of the core provides expertise in a variety of imaging techniques to support the morphological needs to individual projects. These technologies cover the areas of conventional light microscopy, phase contrast microscopy, fluorescence microscopy, laser scanning confocal microscopy (real time and standard 3-dimensional reconstruction of fixed tissues), digital imaging, transmission electron microscopy, in-situ hybridization, immunoohistochemistry and immuno-electron microscopy (immunogold and diaminobenzidine reactions). Other areas that the core has recently developed are calcium imaging of ICC-MY networks using fluo-4, coupled with real time confocal microscopy. The core also provides support in the area of spatio-temporal mapping of excitability within the gastrointestinal tract. The morphology core will also be able to provide FRET analysis in the near future to projects interested in analyzing the interactions of proteins within the plasma membrane or cytoplasm. The quality of the output of the core can be assessed by the images on several front covers of papers that were published in Gastroenterology and American Journal of Physiology.