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A Freeze-Fracture & Etching System for Northwestern Univ

冻裂

基本信息

批准号：
6439852
负责人：
MARK JOHNSON
金额：
$ 20.87万
依托单位：
NORTHWESTERN UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2002
资助国家：
美国
起止时间：
2002-07-01 至 2004-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6439852
关键词：
biomedical equipment purchase cryoelectron microscopy freeze etching

项目摘要

The requested funds will be used to purchase a Freeze-Fracture and Etching System that will be the cornerstone instrument of a new Cryopreparation Facility to be housed in the Electron Probe Instrumentation Center at Northwestern University. We currently have an old freeze-fracture and etching system, but it is limited to a small user base, has limited capacity to process samples, is limited in resolution, is subject to breakdown, and is generally difficult to use. This has limited the potential for studies involving cryopreparation at Northwestern University. The new device has a much greater capability to process samples, is much easier to use and thus will facilitate a much larger user base, and should be less prone to breakdown. The device to be purchased is a Cressington CFE Freeze- Fracture Replication System outfitted with low and 45 degree angle platinum/carbon and tantalum/tungsten shadowing sources, two 90 degree carbon backing evaporation guns, two microtome holders, two Heuser mount specimen stages, one gold hat specimen stage, one double replica stage, two quad Heuser mount stages, low and high angle quartz crystal thickness monitors, dewar pumping and shield kit for ultra-low temperature coating. This system will allow freeze- fracture and freeze-etching of specimens, coating with both platinum/carbon and tantalum/tungsten at low and high angles, high throughput (4 times higher than current device and up to 50 runs or 200 specimens without venting) and coating at ultralow temperature to achieve nearly "grainless" PLATINUM/CARBON COVERAGE. In combination with the different methods of freeaing that will be available (a Med-Vac helium slam freezer and a Leica CPC Universal cryoworkstation), this will provide a versatile facility for cryopreparation. This equipment will be used in a variety of studies including examination of the changes in extracellular matrix architecture in glaucoma, age-related macular degeneration, and lymphangiogenesis, characterization of a variety of biomimetic/bioinspired biomaterials, ultrastructural examination of the intracellular matrix of vascular endothelial cells, characterization of synaptic junctions, detailed examination of the interaction of biomimetic polymeric scaffolds with ovarian follicles and ovarian somatic cells with oocyte, determining the polymorphic phase of cationic and anionic mixtures, investigating the morphology of fused lipid bilayer vesicles, and analyzing the supramolecular organization of the actin machinery in the motility of crawling cells.

申请的资金将用于购买冷冻断裂和蚀刻系统，该系统将作为西北大学电子探针仪器中心新的冷冻修复设施的基石仪器。我们目前有一个旧的冷冻断裂和蚀刻系统，但它限于较小的用户群，处理样品的能力有限，分辨率有限，容易崩溃，通常很难使用。这限制了西北大学涉及冷冻修复的研究的潜力。新设备具有更大的处理样品的能力，更容易使用，因此将促进更大的用户基础，应该不太容易出现故障。将购买的设备是Cressington CFE冷冻断口复制系统，配备有低角度和45度角的铂/碳和钽/钨阴影源，两支90度碳背蒸发枪，两个切片机支架，两个Heuser安装样品台，一个金帽样品台，一个双复制工作台，两个四个Heuser安装工作台，低和高角度石英晶体厚度监测器，杜瓦泵和超低温涂层屏蔽套件。该系统将允许样品的冷冻断裂和冷冻蚀刻，在低角度和高角度用铂/碳和钽/钨进行涂层，高吞吐量(比目前的设备高4倍，最多50次或200个样品不透气)，并在超低温下进行涂层，以实现几乎“无颗粒”的铂/碳覆盖。与可用的不同冷冻方法(Med-Vac氦冷冻机和徕卡CPC通用冷冻工作站)相结合，这将为冷冻修复提供一种多功能的设施。该设备将用于一系列研究，包括检测青光眼、老年性黄斑变性和淋巴管生成过程中细胞外基质结构的变化，各种仿生/生物启发生物材料的表征，血管内皮细胞细胞内基质的超微结构检测，突触连接的表征，仿生聚合物支架与卵巢滤泡和卵巢体细胞与卵母细胞相互作用的详细研究，确定阳离子和阴离子混合物的多晶相，研究融合的脂质双层囊泡的形态，以及分析爬行细胞运动中肌动蛋白机械的超分子组织。