TEM with Digital and Cryopreparation Systems

配备数字和冷冻制备系统的 TEM

• 批准号: 6576822
• 负责人: ANDREW P EVAN
• 金额: $ 38.66万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-06-01 至 2004-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6576822
• 关键词: biomedical equipment purchase; biomedical equipment resource; cell biology; cryoelectron microscopy; cytology; digital imaging; immunocytochemistry; transmission electron microscopy

DESCRIPTION (provided by applicant): The purpose of the application is to obtain funding to replace the existing Philips 400 TEM in the shared-user EM Center in the Department of Anatomy and Cell Biology with a state-of-the-art TEM, the JEOL 1230. The Philips 400 TEM was purchased with NSF funds in 1979 and has served over 40 different researchers over the last 23 years, but this machine is presently unreliable and outdated. Since the acquisition of the Philips 400 TEM, significant technical advances have occurred in transmission electron microscopes which include ease of use and extended capabilities like digital image recording and ultracryo-microscopy. The new TEM will be housed in a 1789 sq. ft. laboratory dedicated to electron microscopy, supervised by a full-time faculty member, Dr. Vincent Gattone and managed by a full-time, highly qualified EM specialist, Ms. Caroline Miller. The microscope will be available to all investigators within the Indiana University-Purdue University campus at Indianapolis which includes Schools of Dentistry, Medicine, Science and Engineering & Technology. However, the majority of the research projects will come from ten (10) laboratories. These investigators have been identified as "major users" of the new instrument. Dr. Robert Bacallao will employ a variety of biochemical assays for protein sorting, immunohistochemical and ultrastructural analyses of Golgi complex dysfunction following ischemic injury. Dr. Andrew Evan will use imunohistochemical and digital imaging to precisely correlate sites of renal crystal deposition with known inhibitors of stone formation. Dr. Loren Field will use TEM to correlate structural changes in cardiomyocytes after specific pathological conditions. Dr. Lincoln Ford requires digital TEM images to accurately measure changes in smooth muscle thick filaments during contractility. Dr. Susan Gunst needs digital TEM images to evaluate the cellular mechanisms that regulate the response of airway smooth muscle to mechanical forces generated during breathing. Dr. James McAteer's studies require the high resolution of the TEM to detect subtle damage to the vascular endothelium induced by shock wave lithotripsy. Dr. Bruce Molitoris will employ immunohistochemistry to determine the cellular, biochemical and molecular mechanisms responsible for ischemia induced membrane changes. Dr. Carrie Phillips will use immunogold EM to evaluate the cellular localization of inversin protein in the inv mutant mouse model of PKD. Dr. Zao Xu will use TEM to investigate the nature of cell death, e.g. necrosis or apoptosis, in neostriatum of the rat after transient global schemia. Dr. Richard Gregory will employ immunohistochemistry to examine the role of a Streptococcus mutans 65 kDa fimbrial protein binding as a mechanism for the induction of dental carries.

描述（由申请人提供）： 该申请的目的是获得资金，用最先进的 TEM JEOL 1230 替换解剖学和细胞生物学系共享用户 EM 中心的现有 Philips 400 TEM。 Philips 400 TEM该机器于 1979 年由 NSF 资金购买，在过去 23 年中已为 40 多名不同的研究人员提供服务，但该机器目前不可靠且过时。自购买 Philips 400 TEM 以来，透射电子显微镜取得了重大技术进步，其中包括易用性和数字图像记录和超低温显微镜等扩展功能。新的 TEM 将坐落在一个 1789 平方英尺的电子显微镜实验室中，由全职教员 Vincent Gattone 博士监督，并由全职高素质 EM 专家 Caroline Miller 女士管理。该显微镜将向印第安纳大学-普渡大学印第安纳波利斯分校校园内的所有研究人员开放，其中包括牙科学院、医学院、科学学院以及工程与技术学院。然而，大多数研究项目将来自十 (10) 个实验室。这些研究人员已被确定为新仪器的“主要用户”。 Robert Bacallao 博士将采用多种生化检测方法对缺血性损伤后高尔基复合体功能障碍进行蛋白质分选、免疫组织化学和超微结构分析。 Andrew Evan 博士将使用免疫组织化学和数字成像技术，将肾晶体沉积部位与已知的结石形成抑制剂精确关联起来。 Loren Field 博士将使用 TEM 来关联特定病理条件后心肌细胞的结构变化。 Lincoln Ford 博士需要数字 TEM 图像来准确测量平滑肌粗丝在收缩过程中的变化。 Susan Gunst 博士需要数字 TEM 图像来评估调节气道平滑肌对呼吸过程中产生的机械力的反应的细胞机制。 James McAteer 博士的研究需要高分辨率的 TEM 来检测冲击波碎石术引起的血管内皮的细微损伤。 Bruce Molitoris 博士将采用免疫组织化学来确定导致缺血诱导的膜变化的细胞、生化和分子机制。 Carrie Phillips 博士将使用immunogold EM 来评估inv 蛋白在PKD 突变小鼠模型中的细胞定位。徐早博士将使用 TEM 来研究细胞死亡的本质，例如：短暂性整体缺血后大鼠新纹状体中的坏死或凋亡。 Richard Gregory 博士将采用免疫组织化学来检查变形链球菌 65 kDa 菌毛蛋白结合作为诱导牙携带机制的作用。