CORE--CELL MORPHOLOGY

核心--细胞形态学

• 批准号: 6919793
• 负责人: Peter Bell
• 金额: $ 23.78万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-07-01 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6919793
• 关键词: bioimaging /biomedical imaging; biomedical facility; biotechnology; cell morphology; cellular pathology; cytogenetics; cytology; green fluorescent proteins; histology; image processing; immunocytochemistry; immunoelectron microscopy; in situ hybridization; light microscopy; transfection /expression vector; transmission electron microscopy

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.

细胞形态核心（CMC）将支持授予成员，以实现其研究目标所需的组织学相关工作的各个方面。关键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，例如β-半乳糖苷酶和GFP），通过电子显微镜评估病毒载体评估以及用于病理评估的标准组织学。具体而言，CMC从石蜡，冷冻和塑料包裹的组织中制备切片。定期进行常规和特殊污渍以及凋亡和细胞增殖测定法。使用所有适当的阳性对照和阴性对照使用荧光或酶标记的抗体进行免疫组织化学。原位杂交用于鉴定转导的细胞或检测 病毒使用标记的DNA或RNA探针。光显微镜图像是用数码相机拍摄的，可以以电子方式或盘式发送给研究人员。可以执行图像分析，量化任何目标的染色程度。进行透射电子显微镜（TEM）以评估超微结构水平的组织样品。该技术可以与具有金标记抗体的转基因蛋白的特异性亚细胞定位与免疫染色（免疫电子显微镜）结合使用。对于向量分析，负 病毒制剂的染色用于EM分析。评估病毒载体的平均密度，纯度，均匀性和病毒粒子完整性。由于两种病毒的大小差异，可检测到具有AAV（反之亦然）的腺病毒制剂中的污染。照片在CMC内拍摄，开发和印刷。调查人员可以收到纸质照片或计算机文件，包括书面评估。