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Cell Morphology Core

细胞形态核心

基本信息

批准号：
8376150
负责人：
Peter L Bell
金额：
$ 7.72万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
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/8376150
关键词：
Address Adenoviruses Affect Alkaline Phosphatase Antibodies Apoptosis Biodistribution Biological Assay Cell Proliferation Cells Cellular Morphology Computer software Confocal Microscopy Critiques Detection Electron Microscope Electron Microscopy Ensure Enzymes Equipment Evaluation Fluorescence Resonance Energy Transfer Freezing Galactosidase Gene Expression Gene Transfer Goals Grant Harvest Head Hereditary Disease Histocytochemistry Histological Techniques Histology Histopathology Housing Image Imagery In Situ Hybridization Knowledge Label Laboratories LacZ Genes Leadership Light Microscope Methodology Methods Microscope Microscopic Microscopy Microtome - medical device Molecular Molecular Biology Morphology Negative Staining Organ Paraffin Pathologic Plastic Embedding Play Preparation Process Proteins Protocols documentation RNA RNA Probes Reporter Genes Research Research Personnel Resources Sampling Services Staining method Stains Techniques Tissue Sample Tissues Training Transgenes Translational Research Transmission Electron Microscopy Viral Viral Vector Virus Walking Work base cell type cellular transduction cryostat density experience fluorescence microscope gene therapy instrument member particle programs ranpirnase research study square foot success tissue processing transgene expression transmission process vector viral detection web page

项目摘要

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.

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。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。