CORE--CELL MORPHOLOGY

核心--细胞形态学

• 批准号: 7266266
• 负责人: Peter Bell
• 金额: $ 25.22万
• 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/7266266
• 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，例如β-半乳糖苷酶和GFP）、电子显微镜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、冷冻和塑料包埋组织制备切片。定期进行常规和特殊染色以及细胞凋亡和细胞增殖测定。使用所有适当的阳性和阴性对照，使用荧光或酶标记抗体进行免疫组织化学。原位杂交用于鉴定转导细胞或检测 病毒使用标记的DNA或RNA探针。光学显微镜图像是用数码相机拍摄的，可以通过电子方式或光盘发送给调查人员。可以进行图像分析，量化任何目标的染色程度。进行透射电子显微镜（TEM）以在超微结构水平上评价组织样品。该技术可以与免疫染色（免疫电子显微镜）结合，用于用金标记抗体对转基因蛋白进行特异性亚细胞定位。对于矢量分析，阴性 将病毒制剂染色用于EM分析。评估病毒载体的平均密度、纯度、均匀度和病毒颗粒完整性。由于两种病毒大小的差异，腺病毒制剂中的AAV污染（反之亦然）是可检测的。照片在CMC内拍摄、显影和打印。调查人员可以收到硬拷贝照片或包括书面评价在内的计算机文件。