Cell Morphology Core

细胞形态学核心。宾夕法尼亚大学的基因治疗计划是一个设备齐全的设施，能够进行组织学，显微镜和形态学分析的各个方面。这包括常规的组织学程序，包括尸检以及更先进的技术，如免疫荧光，原位杂交，酶活性染色和电子显微镜。在核心实验室空间内提供了处理和分析组织样本的所有必要仪器（包括 光学显微镜和电子显微镜）。研究人员可以使用核心的显微镜和成像软件从他们的样本中获取图像。 细胞形态学核心将执行所有的组织学和显微镜相关的工作需要为三个项目的赠款。这将包括以下程序：项目1：小鼠尸检; OTC、OTC突变体和MHCI-SIINFEKL的免疫染色; SIINFEKL特异性T细胞的原位四聚体染色。项目二：小鼠尸检;肝脏GFP检测;针对一般病理学（H&E）、纤维化（三色/天狼星红）、脂肪变性（油红-O）、细胞凋亡（TUNEL测定）、炎症（针对CD 4/CD 8 T细胞浸润的免疫染色）的肝脏染色;针对肝脏中hOTC表达的免疫染色; OTC酶 活性染色项目3：小鼠尸检;肝脏中GFP的检测;肝脏中hOTC表达的免疫染色，H&E染色。 细胞形态学核心的优势在于拥有多年的基因治疗研究肝组织分析经验，特别是针对该资助的每个项目提出的分析技术。虽然一些常规技术很容易执行，但某些方法，如通过免疫荧光或酶组织化学检测spf^^“肝脏中的人OTC，需要遵守已建立的 和经过测试的协议。方案已经制定出来，以前成功地用于几乎所有计划的组织学方法，不需要新开发。核心负责人执行所有实验，如免疫染色，并采用严格的对照品，以确认染色和载玻片的特异性，在将其发布给研究者之前，由核心负责人审查。行政核心监督小组 形态核心的预算和审查服务的定价，以确保成本效益。

The Cell Morphology Core at U. Penn's Gene Therapy Program is a fully equipped facility capable to carry out all aspects of histology, microscopy, and morphometric analyses. This includes routine histology procedures including necropsies as well as more advanced techniques such as multicolor immunofluorescence, in situ hybridization, enzyme activity staining and electron microscopy. All necessary instruments to process and analyze tissue samples are available within the Core's lab space (including microscopes for light and electron microscopy). Investigators can use the Core's microscopes and imaging software to acquire images from their samples. The Cell Morphology Core will perform all the histology- and microscopy-related work required for the three projects of this grant. This will include the following procedures: Project 1: mouse necropsies; immunostaining for OTC, OTC mutants, and MHCI-SIINFEKL; in situ tetramer staining for SIINFEKL-specific T cells. Project 2: mouse necropsies; GFP detection in liver; staining of livers for general pathology (H&E), fibrosis (trichrome/Sirius Red), steatosis (oil-red-O), apoptosis (TUNEL assay), inflammation (immunostaining for CD4/CDS T cell infiltrates); immunostaining for hOTC expression in liver; OTC enzyme activity staining. Project 3: mouse necropsies; detection of GFP in liver; immunostaining for hOTC expression in liver, H&E staining. The Cell Morphology Core has the advantage of having many years of experience with analyzing liver tissues for gene therapy studies and in particular with the analytical techniques proposed for each project of this grant. While some routine techniques are easy to perform, certain methods such as detection of human OTC in spf^^'' liver by immunofluorescence or enzyme histochemistry require the adherence to established and tested protocols. Protocols have been worked out and previously used successfully for almost all planned histological methods and do not need to be newly developed. The Core performs all experiments such as immunostains with stringent controls to confirm the specificity of the stains and slides are reviewed by the Core director before being released to the investigator. The Administrative Core oversees the Cell Morphology Core's budget and reviews pricing of services to assure cost effectiveness.