CORE--CELL MORPHOLOGY

核心--细胞形态学

• 批准号: 7457702
• 负责人: Peter Bell
• 金额: $ 26.16万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7457702
• 关键词: Adenoviruses; Antibodies; Apoptosis; Biological Assay; Cell Proliferation; Cellular Morphology; Computers; DNA; Detection; Electron Microscopy; Enzymes; Evaluation; Freezing; Goals; Gold; Grant; Green Fluorescent Proteins; Histology; Image; Image Analysis; Immunoelectron Microscopy; Immunohistochemistry; In Situ Hybridization; Label; Light; Microscopic; Negative Staining; Paraffin; Plastic Embedding; Preparation; Printing; Proteins; RNA Probes; Reporter Genes; Research; Research Personnel; Services; Staining method; Stains; Standards of Weights and Measures; Techniques; Tissue Sample; Tissues; Transgenes; Transgenic Organisms; Transmission Electron Microscopy; Viral; Viral Vector; Virus; Work; Writing; beta-Galactosidase; cellular transduction; density; digital; member; particle; size; vector; viral detection

The Cell Morphology Core (CMC) will support the grant members with all aspects of histology-related work necessary to accomplish their research goals. Key services will be transgene detection in tissues (immunohistochemistry, in situ hybridization, reporter gene detection such as beta-galactosidase and GFP), viral vector evaluation by electron microscopy, and standard histology for pathological evaluation. Specifically, The CMC prepares sections from paraffin, frozen and plastic-embedded tissues. Routine and special stains, as well as apoptosis and cell proliferation assays, are performed regularly. Immunohistochemistry is performed with fluorescent or enzyme-labeled antibodies using all the appropriate positive and negative controls. In situ hybridization for identification of transduced cells or the detection of viruses uses labeled DNA or RNA probes. Light microscopic images are taken with a digital camera and can be sent to investigators electronically or on a disc. Image analysis can be performed, quantifying the degree of staining for any target. Transmission electron microscopy (TEM) is performed to evaluate tissue samples at an ultrastructural level. This technique can be combined with immunostaining (immuno-electron microscopy) for the specific subcellular localization of transgenic proteins with gold-labeled antibodies. For vector analysis, negative staining of viral preparations is applied for EM analysis. Virus vectors are assessed for average density, purity, uniformity and viral particle integrity. Contaminations in adenovirus preparations with AAV (and vice versa) are detectable due to the difference in size of both viruses. Photographs are taken, developed and printed within the CMC. Investigators can receive either the hard copy photographs or computer files including written evaluations.

细胞形态学核心（CMC）将支持资助成员完成其研究目标所需的组织学相关工作的各个方面。主要服务将是组织中的转基因检测（免疫组织化学，原位杂交，报告基因检测，如β -半乳糖苷酶和绿色荧光蛋白），电子显微镜下的病毒载体评估，以及病理评估的标准组织学。具体来说，CMC从石蜡、冷冻和塑料包埋的组织中制备切片。常规和特殊染色，以及细胞凋亡和细胞增殖试验，定期进行。免疫组织化学用荧光或酶标记抗体使用所有适当的阳性和阴性对照进行。原位杂交鉴定转导细胞或检测