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Cell Morphology Core

细胞形态核心

基本信息

批准号：
9060373
负责人：
Peter Bell
金额：
$ 9.36万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至 2018-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9060373
关键词：
Adenovirus Vector Adherence Apoptosis Autopsy Basic Science Biological Assay Budgets Cellular Morphology Clinic Clinical Contracts Control Groups Detection Development Disease Electron Microscopy Enzymes Equipment Evaluation Fibrosis Generations Grant Guidelines Hematoxylin and Eosin Staining Method Hereditary Disease Histocytochemistry Histology Human Image Immunofluorescence Immunologic In Situ In Situ Hybridization Inflammation Instruction Laboratories Lentivirus Vector Liver Measures Methods Microscope Microscopy Mus Oils Ornithine carbamoyltransferase deficiency Pathology Performance Phase Price Procedures Process Production Protocols documentation Quality Control Regulation Research Research Personnel Resources Sampling Serotyping Services Slide Specificity Staining method Stains T-Lymphocyte TdT-Mediated dUTP Nick End Labeling Assay Techniques Technology Testing Tissue Sample Tissues Translations Trichrome stain method Viral Vector Work adeno-associated viral vector base cost effective cost effectiveness design enzyme activity experience gene therapy gene transfer vector good laboratory practice imaging software instrument interest light microscopy meetings microscopic imaging mutant novel pre-clinical preclinical study programs research study therapeutic development urea cycle vector

项目摘要

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.

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、显微镜和形态计量学分析的各个方面。这包括常规的组织学程序，包括尸检，以及更先进的技术，如多色免疫荧光，原位杂交，酶活性染色和电子显微镜。所有处理和分析组织样本所需的仪器都可以在Core的实验室空间中使用(包括