TEM with Digital and Cryopreparation Systems

描述（由申请人提供）： 该申请的目的是获得资金，以取代现有的飞利浦400 TEM在解剖学和细胞生物学系的共享用户EM中心与最先进的TEM，JEOL 1230。Philips 400 TEM于1979年由NSF基金购买，在过去的23年中为40多名不同的研究人员提供了服务，但这台机器目前不可靠且过时。自从收购Philips 400 TEM以来，透射电子显微镜取得了重大技术进步，包括易用性和扩展功能，如数字图像记录和超低温显微镜。新的TEM将被安置在一个1789平方米。英尺实验室致力于电子显微镜，由全职教师，博士文森特Gattone监督和管理的全职，高素质的EM专家，女士卡罗琳米勒。该显微镜将提供给位于印第安纳波利斯的印第安纳州普渡大学校园内的所有研究人员，其中包括牙科，医学，科学和工程与技术学院。然而，大多数研究项目将来自十（10）个实验室。这些调查人员被确定为新仪器的“主要用户”。Robert Bacallao博士将采用各种生化分析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地联系起来。Loren Field博士将使用TEM来关联特定病理条件下心肌细胞的结构变化。林肯福特博士需要数字TEM图像，以准确测量收缩过程中平滑肌粗丝的变化。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TEM的高分辨率来检测冲击波碎石术对血管内皮的细微损伤。布鲁斯莫利托里斯博士将采用免疫组织化学来确定负责缺血诱导膜变化的细胞、生化和分子机制。Dr. Carrie菲利普斯将使用免疫金EM评价PKD inv突变小鼠模型中反转蛋白的细胞定位。Zao Xu博士将使用TEM研究短暂性全脑缺血后大鼠新纹状体中细胞死亡的性质，例如坏死或凋亡。Richard Gregory博士将采用免疫组织化学方法研究变形链球菌65 kDa菌毛蛋白结合作为诱导牙齿携带机制的作用。

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1979 and has served over 40 different researchers over the last 23 years, but this machine is presently unreliable and outdated.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.