Cell Morphology Core

宾夕法尼亚大学基因治疗项目的细胞形态学核心是一个设备齐全的设施，能够进行组织学、显微镜和形态分析的各个方面。这包括常规组织学程序，包括尸检以及更先进的技术，例如多色免疫荧光、原位杂交、酶活性染色和电子显微镜。核心实验室空间内提供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 Core 的显微镜和成像软件从样本中获取图像。 细胞形态学核心将执行本次资助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：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2：小鼠尸检；肝脏中GFP检测；肝脏染色以了解一般病理学 (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（CD4/CDS T 细胞浸润的免疫染色）；肝脏 hOTC 表达的免疫染色；非处方酶 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分析肝组织以进行基因治疗研究的经验，特别是针对本次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（例如通过免疫荧光或酶组织化学检测 spf^^'' 肝脏中的人 OTC）需要遵守已建立的规则 并测试了协议。几乎所有计划的组织学方法都已经制定并成功使用了协议，不需要重新开发。 Core 执行所有实验，例如免疫染色，并进行严格的控制，以确认染色剂的特异性，载玻片在发布给研究者之前由 Core 主管进行审查。行政核心负责监督该小组 Morphology Core 的预算并审查服务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.