CORE--DI VIRAL VECTOR

核心--DI病毒载体

• 批准号: 6302740
• 负责人: MICHEAL M LAI
• 金额: $ 18.61万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-04-01 至 2001-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6302740
• 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.

神经病理学核心为所有四个项目提供必要的服务。 AD项目中详细的组织病理学改变，无论是由于DI 新基因的载体驱动表达或感染分析 免疫缺陷小鼠，是许多建议的关键终点 实验。组织样本由岩心获取，作为 实验程序。动物很少会专门用来 获取该岩心的样品。例外的是这些程序 (塑料包埋和EM)，需要灌流。组织次数 获取由单独的实验决定，并且是协调的 由核心主管负责。核心研究助理负责 所有组织的加工、切片和染色。例行检查， 在大多数动物上进行的包括H&E、豪华坚牢蓝 用髓鞘和免疫过氧化物酶测定病毒的分布 AG.针对特定亚型细胞的特殊染色，表达 细胞因子、原位杂交等是由个体决定的 实验也受到质量控制，并与计划一起进行 和核心导演在一起。EPON嵌入材料和冷冻选择是 也由内核处理。我们很幸运地保留了这些服务 在过去的8年里。他在所有提议的 技术，并一直是我们正在进行的研究的无价资产 努力。根据实验的性质，幻灯片可以是 由核心导演筛选或由希尔顿博士盲读 或者两者都有。对关键实验的盲读消除了调查人员 偏见。 在过去，我们已经为所有提议的 技术，因此预计不会有任何困难。一种协议 值得注意的变化是使用指定为RIP的鼠单抗 少突胶质细胞染色。该单抗对少突胶质细胞和 可识别的大鼠和小鼠来源的少突胶质细胞，可用于 在福尔马林固定或冰冻切片上。含RIP单抗的上清液 是由Regeneron公司提供的，因为他们无法为我们提供 与杂交瘤直接接触。