CORE--CELL MORPHOLOGY

细胞形态学核心 (CMC) 将为资助成员提供实现其研究目标所需的组织学相关工作的各个方面的支持。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，如β-半乳糖苷酶和GFP）、通过电子显微镜评估病毒载体以及用于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。定期进行常规和特殊染色以及细胞凋亡和细胞增殖测定。使用荧光或酶标记抗体并使用所有适当的阳性和阴性对照进行免疫组织化学。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研究人员。可以进行图像分析，量化任何目标的染色程度。透射电子显微镜 (TEM) 用于在超微结构水平上评估组织样本。该技术可以与免疫染色（免疫电子显微镜）相结合，用金标记抗体对转基因蛋白进行特定的亚细胞定位。对于矢量分析，负 病毒制剂的染色用于电镜分析。评估病毒载体的平均密度、纯度、均匀性和病毒颗粒完整性。由于两种病毒大小的差异，可以检测到 AAV 腺病毒制剂中的污染（反之亦然）。照片在 CMC 内拍摄、冲洗和打印。调查人员可以收到硬拷贝照片或包括书面评估的计算机文件。

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.