IMAGING TRANSMISSION ELECTRON MICROSCOPE

最近克隆的蛋白质的高分辨率电子显微镜免疫标记现在可以清楚地识别在神经和神经内科疾病中影响大脑功能的部位。这种独特的能力，再加上对定量超微结构数据的更大需求，使得康奈尔大学威尔医学院(WMC-CU)神经病学、神经科学和药理学系的教职员工对现代透射电子显微镜(TEM)的使用产生了前所未有的需求。该小组的原始研究主要使用1973年的飞利浦201透射电子显微镜模型提供了关于神经递质转运体和受体的亚细胞分布的基本信息，该模型已过时，无法经济有效地满足研究人员的需求。该提案要求购买一台新的透射电子显微镜(Tecnai 12 BioTwin)，配备数码相机，并具有在线成像和定量分析的能力。这种技术先进的系统的使用将确保完成由美国国立卫生研究院几个不同分支机构资助的重大生物医学研究项目。这些资金不能从任何一名研究人员的研究拨款中获得，除非通过NIH共享仪器计划才能获得。本提案中的主要用户群体--六名主要调查人员(S)--的资助项目完全依靠瞬变电磁法进行数据分析。这些研究需要使用透射电子显微镜来定量确定正常的亚细胞分布以及药物或病变诱导的阿片受体和其他功能蛋白在大鼠中枢神经系统(CNS)内靶向的变化。组成少数使用者群体的其他六名研究人员的项目需要对中枢神经系统抗原进行更有限但关键的在线电子显微镜分析，特别是在神经退行性疾病和药物成瘾的遗传动物模型中。总之，这些研究的结果将对理解人类的疼痛控制、自主神经功能、动机行为和记忆具有重要意义。为了确保新的瞬变电磁系统的长期成功使用，仪器的日常操作将由一个由所有主要用户组成的内部咨询委员会管理，并由经验丰富的工作人员在医学院的全力支持下进行维护。

High resolution electron microscopic immunolabeling of recently cloned proteins now permits clear identification of sites mediating brain functions that are affected in neurological and neurophychiatric diseases. This unique capability, together with a greater demand for quantitative ultrastructural data, has produced an unprecedented need for the use of a modern transmission electron microscope (TEM) by the faculty within the Departments of Neurology and Neuroscience and Pharmacology at Weill Medical College of Cornell University (WMC-CU). Original studies from this group have provided fundamental information on the subcellular distribution of neurotransmitter transporters and receptors using primarily a 1973 model of a Philips 201 TEM, which has become obsolete and unable to cost-effectively meet the needs of the investigators. This proposal requests funds to purchase a new TEM (Tecnai 12 BioTwin) with a digital camera and capacity for on-line imaging and quantitative analysis. The availability of this technologically advanced system will assure the completion of major biomedical research projects that are funded from several different branches of the NIH. These funds cannot be obtained from the research grants of any one investigator, and are not available except through the NIH Shared Instrument Program. The funded projects of the six principal investigators (P.I.'s) who comprise the major users group in this proposal rely exclusively on a TEM for data analysis. These studies require the use of a TEM for quantitatively determining the normal subcellular distribution and drug- or lesion-induced changes in the targeting of opioid receptors and other functional proteins within the rat central nervous system (CNS). The projects of six other investigators who comprise the minor users group require more limited, but crucial, on-line TEM analysis of CNS antigens, particularly in genetic animal models of neurodegenerative diseases and drug addiction. Together, the results from these studies will have major implications for understanding the control of pain, autonomic functions, motivated behaviors, and memory in humans. To assure the successful long-term use of the new TEM system, the daily operation of the instrument will be managed by an internal advisory committee of all major users, and maintained by an experienced staff with the full support of the Medical College.