CORE--DI VIRAL VECTOR

核心--DI病毒载体

• 批准号: 6585583
• 负责人: MICHEAL M LAI
• 金额: $ 10.62万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-04-01 至 2003-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6585583
• 关键词: biomedical facility; confocal scanning microscopy; immunopathology; murine hepatitis virus; transfection /expression vector; virus RNA; virus protein

The neuropathology core provides services essential to all four projects. Ad detailed in the projects histopathological changes, whether due to DI vector driven expression of novel genes or analysis of infection in immunodeficient mice, is the critical end point for many of the proposed experiments. Tissue specimens are acquired by the core as part of the experimental procedures. Animals are rarely dedicated solely to acquisition of samples for this core. Exceptions are those procedures (plastic embedding and EM) which require perfusion. Times of tissue acquisition are dictated by the individual experiments and are coordinated by the core director. The core research associate is responsible for processing, sectioning and staining of all tissues. Routine examinations, carried out on the majority of animals include H&E, luxol fast blue of myelin and immunoperoxidase determinations for the distribution of viral Ag. Special stains for specific subtypes of cells, expression of cytokines, in situ hybridizations etc., are dictated by the individual experiments and are also quality controlled and schedule in conjunction with the core director. Epon embedded materials and frozen selections are also processed by the core. We are fortunate to have retained the services of Wen for the past 8 years. He has experience in all of the proposed techniques and has been an invaluable asset to our ongoing research efforts. Depending upon the nature of the experiments, slides are either screened by the core director or read in a blinded fashion by Dr. Hilton or both. Blinded reading of critical experiments eliminates investigator bias. In the past we have published precise protocols for all of the proposed techniques and therefore no difficulties are anticipated. One protocol change worthy of note is the use of the murine mAb designated RIP for staining of oligodendroglia. This mAb is specific for oligodendroglia and recognized oligodendroglia of both rat and mouse origins and can be used on formalin fixed or frozen sections. Supernatants containing the RIP mAb are kindly provided by Regeneron Inc. since they are unable to provide us directly with the hybridoma.

神经病理学核心提供对所有四个项目至关重要的服务。 详细说明项目中的组织病理学变化，是否由于DI 载体驱动的新基因表达或感染分析， 免疫缺陷小鼠，是许多建议的关键终点。 实验组织标本由核心采集，作为 实验程序。动物很少会专门 采集该岩心的样品。这些程序 （塑料包埋和EM），需要灌注。组织次数 采集由单独的实验决定并进行协调 核心导演。核心研究助理负责 所有组织的处理、切片和染色。常规检查， 在大多数动物身上进行的测试包括H&E，luxol坚牢蓝， 髓鞘和免疫过氧化物酶测定用于病毒分布 AG.针对特定细胞亚型的特殊染色， 细胞因子，原位杂交等，是由个人决定的 实验，也是质量控制和时间表结合 和核心主管Epon嵌入材料和冷冻选择是 也由核心处理。我们很幸运能保留这些服务 在过去的8年里，他在所有建议的经验， 技术，并已成为我们正在进行的研究的宝贵资产 努力根据实验的性质，载玻片可以是 由核心主任筛选或由希尔顿博士盲读 或两者关键实验的盲态阅读排除了研究者 bias. 在过去，我们已经发布了所有建议的精确协议， 技术，因此预计没有困难。一个协议 值得注意的变化是使用命名为RIP的鼠mAb用于 少突胶质细胞染色。该mAb对少突胶质细胞具有特异性， 识别的大鼠和小鼠来源的少突胶质细胞， 福尔马林固定或冷冻切片。含RIP mAb的上清液 由Regeneron Inc.因为他们无法向我们提供 直接用杂交瘤。