JEOL Freeze Fracture/Freeze Etch Device

JEOL 冷冻断裂/冷冻蚀刻装置

• 批准号: 8640669
• 负责人: DONNA BEER STOLZ
• 金额: $ 30.02万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-05-01 至 2015-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8640669
• 关键词: 20 year old; Area; Country; Coupled; Devices; Electron Microscopy; Electrons; Equipment; Faculty; Freeze Etching; Freeze Fracturing; Funding; Guns; Image; Image Analysis; Institution; Label; Light; Membrane; Methods; Microscopy; Modality; Molecular; Nature; Protocols documentation; Research; Research Personnel; Resolution; Shadowing (Histology); Structure; Techniques; Technology; Time; United States National Institutes of Health; Universities; Vacuum; instrument; light microscopy; medical schools; member; optical imaging

The use of freeze-fracture, deep-etch and molecular replica methods are currently finding a tremendous resurgence of utility in modern electron microscopy. Regardless of the increased sophistication of cryo- methods and other advanced EM technologies and coincident improvements in the resolution of light microscopy, traditional EM microscopies still have incredible utility. Moreover, as many facilities have been closed with the advent of high-resolution light microscopy, those that remain have become increasingly, and notably in our case, phenomenally active, collaborating with research groups from all over the country. The cause of the re-emergence in the use of replica methods springs from the need to localize molecules at higher resolution in membranes. Traditionally, we have significant expertise in the technique, and while the expertise still exists, the equipment in our facility does not. Our freeze-fracture machine, a refurbished ~20 year old Cressington CFE50, was retired two years ago due to vacuum, gun and electrical problems that could not be remedied by the company since this specific instrument is no longer manufactured. Because of this, we are requesting funds to purchase a JEOL JFD-V-TP freeze-fracture/freeze-etch/rotary shadowing machine for the Center for Biologic Imaging (CBI) at the University of Pittsburgh Medical School. The reason for our need to continue the use of these protocols by purchasing a new instrument is four-fold: 1. The recent recruitment of several new faculty members that have critical requirements for technology afforded by this instrument, including the rotary shadowing component of the machine that cannot be purchased as a stand alone feature. 2. Ongoing requests from long time collaborators for access to high-resolution structure-function imaging of membranes and molecular replicas of molecules. 3. Integration of light microscopy modalities coupled with high-resolution analysis at the membrane, especially utilizing FRIL (Freeze-Fracture Replica Immuno- Labeling). 4. There are no other devices of this nature anywhere at the University of Pittsburgh, or in the greater Pittsburgh area, for that matter. The mandate of the CBI is to provide access to a full range of light and electron optical, image analysis, and morphometric methods to all research groups (over 250 NIH-funded users) within the University of Pittsburgh as well as many from outside institutions. Acquisition of this machine will assist our major users enormously and provide all other researchers access to important high-resolution technologies to further their research.

冷冻断裂、深蚀刻和分子复制方法的使用目前发现了巨大的 现代电子显微镜的复兴。尽管低温技术越来越复杂， 方法和其他先进的EM技术，并同时提高了光的分辨率 传统的EM显微镜仍然具有令人难以置信的实用性。此外，由于许多设施已 随着高分辨率光学显微镜的出现而关闭，那些仍然存在的已经变得越来越多， 特别是在我们的案例中，非常活跃，与来自全国各地的研究小组合作。的 使用复制方法重新出现的原因是需要在更高的温度下定位分子。 膜的分辨率。传统上，我们在该技术方面具有重要的专业知识，而专业知识 我们的设备还在，但我们的设备不在。我们的冷冻断裂机，翻新~20岁 Cressington CFE 50两年前因真空、喷枪和电气问题而退役，无法使用 该公司已采取补救措施，因为该特定仪器已不再生产。因此，我们 申请资金购买JEOL JFD-V-TP冷冻断裂/冷冻蚀刻/旋转阴影机， 匹兹堡大学医学院生物成像中心（CBI）。我们之所以需要 通过购买新仪器继续使用这些协议有四个方面：1.最近招聘的 几位新教员对该仪器提供的技术有着关键的要求， 包括不能作为独立特征购买的机器的旋转阴影部件。 2.长期合作者持续要求获得高分辨率结构-功能成像， 膜和分子的分子复制品。3.光学显微镜模式的集成，以及 在膜上进行高分辨率分析，特别是利用FRIL（冷冻断裂免疫分析）， 标签）。4.在匹兹堡大学的任何地方都没有这种性质的其他设备， 大匹兹堡地区，就此而言。CBI的任务是提供全方位的照明， 电子光学，图像分析和形态测量方法的所有研究小组（超过250个NIH资助的 用户）以及许多来自外部机构的匹兹堡大学。购买这台机器 将极大地帮助我们的主要用户，并为所有其他研究人员提供重要的高分辨率 技术，以进一步研究。