PURCHASE OF CRYO-SYSTEM (BALZERS HPF010 AND BAF060 FFU)

冷冻固定化细胞和组织提供了最佳的方法 细胞黏附分子(CAM)在细胞表面的结构保持 微生物和细胞的胞外表面。请拨出资金以 购买由a)Balzers高压冷冻机组成的冷冻系统 010，以及b)Balzers冷冻压裂装置(BAF 060)。这个冷冻系统将 用于高压固定化大细胞或细胞团 在2100大气压的液氮中冷冻，然后将它们插入 冷冻转移到Balzers FFU 060的胞外表面 这些细胞将通过深度蚀刻暴露出来，以揭示CAM。营地将会是 通过使用BAF060沉积一层薄金属膜来复制 无论是单向阴影还是双角度旋转阴影，然后 用5纳米的碳层稳定金属膜。金属薄膜 (细胞表面)将由高分辨率场成像 使用Autrata改进型背散射电子探测器的发射扫描电子显微镜 成像胶片，可能还有胶体金标签，要灵敏 原子序数对比度。这些设备将被安置在 细胞生物学和神经解剖学，一个可供 明尼苏达大学校区。主要用户将有权保证访问 至少75%的设备时间和休息时间将提供给其他 基于内部咨询委员会制定的政策的用户。这 冷冻系统将允许我们继续研究检测到 细菌和哺乳动物细胞上的单个CAM。主要用户将申请 这项技术与冷冻扫描或透射电子显微镜一起用于人体的研究 白细胞、血小板、蛋白质附着在成熟的表面 精子，与接合和附着有关的细菌相互作用 肠上皮细胞，以及包括软骨在内的水合组织 还有胶原蛋白垫子。

Cryoimmobilization of cells and tissues offers the best method for the structural preservation of cell adhesion molecules (CAM) on the extracellular surfaces of microorganisms and cells. Funds are requested to purchase a cryosystem composed of a) Balzers High Pressure Freezer Machine 010, and b) Balzers Freeze Fracture Unit (BAF 060). This cryosystem will be used to immobilize large cells or aggregates of cells by high pressure freezing at 2100 atmosphere of liquid nitrogen and then inserting them by cryotransfer into the Balzers FFU 060 where the extracellular surfaces of the cells will be exposed by deep etching to reveal CAM. The CAMP will be replicated by depositing a thin metal film using the BAF 060 to make either unidirectional shadowing of dual angle rotary shadowing, and then stabilizing the metal film with a 5 nm layer of carbon. The metal film (topographical surface of cell) will be imaged by high resolution field emission SEM using an Autrata modified back-scatter electron detector to image the film, possibly colloidal gold labels as well, be sensitive atomic number contrast. This equipment will be housed in the Department of Cell Biology and Neuroanatomy, a facility available to the entire University of Minnesota campus. Major users will have guaranteed access to at least 75% of equipment time and rest will be made available to other users based on policies set by an Internal Advisory Committee. This cryosystem will permit us to continue investigations on the detection of individual CAM on bacteria and mammalian cells. Major users will apply this technology together with cryoSEM or TEM to the study of human leukocytes, platelets, attachment of proteins to the surface of maturing sperm, bacterial interaction related to conjugation and attachment to intestinal epithelial cells, and to hydrated tissues including cartilage and collagen mats.