Cell Morphology Core

宾夕法尼亚大学基因治疗计划的细胞形态核心是一个设备齐全的设施，能够进行组织学、显微镜和形态计量分析的所有方面。这包括常规的组织学程序，包括尸检以及更先进的技术，如多色免疫荧光、原位杂交、酶活性染色和电子显微镜。处理和分析组织样本的所有必要仪器都在核心的实验室空间内(包括 光学显微镜和电子显微镜)。研究人员可以使用Core的显微镜和成像软件从他们的样本中获取图像。 细胞形态核心将执行这笔赠款的三个项目所需的所有组织学和显微镜相关工作。这将包括以下程序：项目1：老鼠身体解剖； 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2：小鼠身体解剖；肝脏GFP检测；肝脏常规病理(H&E)、纤维化(三色/天狼星红)、脂肪变性(油红O)、细胞凋亡(TUNEL法)、炎症(CD4/CDS T细胞浸润免疫染色)；肝脏Hotc表达的免疫染色；OTC酶 活性染色。项目3：小鼠身体解剖；肝脏绿色荧光蛋白检测；肝脏Hotc表达免疫染色，H&E染色。 细胞形态核心的优势是在基因治疗研究的肝组织分析方面拥有多年的经验，特别是在为这笔赠款的每个项目提出的分析技术方面。虽然一些常规技术很容易执行，但某些方法，如免疫荧光或酶组织化学检测SPF^‘’肝脏中的人类OTC，需要遵守已建立的 和测试过的协议。方案已经制定出来，并且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。核心执行所有的实验，如严格控制的免疫染色，以确认染色的特异性，并在发放给研究人员之前，由核心主管审查载玻片。管理核心对单元进行监督 形态核心的预算和审查定价的服务，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.