Cell Morphology Core

细胞形态核心(CMC)为资助成员提供与组织学相关的所有方面的工作 完成他们的研究目标所必需的。核心由彼得·贝尔博士指导，安置在800号楼 一平方英尺的专用空间。它拥有处理组织和细胞的所有必要设备 一系列的组织学分析。这包括几个荧光显微镜，一个传输电子 显微镜、显微镜、超微切割机、几台低温恒温器和自动组织处理设备。 对于光学显微镜水平的组织分析，使用石蜡、冷冻和塑料包埋的样品。 常规和特殊染色以及组织中报告基因表达的检测(GFP，LacZ)是 定期演出。进一步的技术包括免疫染色、原位杂交、细胞凋亡和细胞 细胞增殖实验。免疫染色是用荧光或酶标记抗体进行的，使用所有 适当的积极对照和消极对照。原位杂交用于鉴定转导细胞或 病毒的检测使用标记的DNA或RNA探针。图像可以通过宽视场或共焦方式获取 显微镜观察并用Image-Pro Plus和分析软件分析，量化染色程度 任何目标。 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。 对于媒介分析，病毒制剂的负染用于EM分析。病毒载体是 评估平均密度、纯度、均匀性和病毒颗粒完整性。腺病毒中的污染物 由于两种病毒的大小不同，携带AAV的制剂(反之亦然)是可以检测到的。 调查人员还可以接受组织学技术和显微镜方面的培训，使他们能够 他们利用核心的资源进行自己的分析。

The Cell Morphology Core (CMC) supports the grant members with all aspects of histology-related work necessary to accomplish their research goals. The core is directed by Dr. Peter Bell and is housed in 800 square feet of dedicated space. It owns all the equipment necessary to process tissues and cells for a wide array of histological analyses. This includes several fluorescence microscopes, a transmission electron microscope, microtomes, an ultramicrotome, several cryostats, and automated tissue processing equipment. For tissue analysis at the light microscope level, paraffin, frozen and plastic-embedded samples are used. Routine and special stains as well as the detection of reporter gene expression in tissues (GFP, LacZ) are performed regularly. Further techniques include immunostaining, in situ hybridization, apoptosis and cell proliferation assays. Immunostaining 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 DMA or RNA probes. Images can be acquired by widefield or confocal microscopy and analyzed with Image-Pro Plus and AnalySIS software, quantifying the degree of staining for any target. Transmission electron microscopy (TEM) is performed to evaluate tissue samples at an ultrastructural level. For vector analysis, negative staining of viral preparations is applied for EM analysis. Viral 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. Investigators can also receive training in histological techniques and microscopy allowing them to perform their own analyses by utilizing the core's resources.