CORE--CELL MORPHOLOGY

细胞形态核心(CMC)将为资助成员提供与组织学相关的所有方面的工作，以实现他们的研究目标。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)，电子显微镜病毒载体评估，以及病理评估的标准组织学。具体地说，CMC准备石蜡、冰冻和塑料包埋组织的切片。常规和特殊染色，以及细胞凋亡和细胞增殖分析，定期进行。使用所有适当的阳性和阴性对照，用荧光或酶标记抗体进行免疫组织化学。原位杂交法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。利用透射电子显微镜在超微结构水平上对组织样品进行评估。这项技术可以与免疫染色(免疫电子显微镜)相结合，对带有金标抗体的转基因蛋白进行特定的亚细胞定位。对于向量分析，为负值 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、一致性和病毒颗粒的完整性。由于两种病毒的大小不同，可以检测到带有AAV的腺病毒制剂中的污染(反之亦然)。照片是在CMC内拍摄、冲洗和打印的。调查人员可以收到硬拷贝照片或包括书面评估在内的计算机文件。

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.