Acquisition of a Freeze Fracture Instrument and a High Pressure Freezer

这是一个建议资金购买冻结骨折 仪器和高压冷冻机。 冷冻断裂 机器需要确定：（1）类型和 蛋白质在膜中的分布;（2）囊泡的形态 融合;（3）细胞外基质 （4）非双层脂质在膜中的作用 功能 该研究包括分析自然和 人工膜，对照和实验的比较 样品，并调查植物和原生生物。 （未成年人用户） 还将研究真菌、动物和DNA分子。 冻结 骨折，这涉及分裂和复制细胞， 膜，是唯一的技术， 膜内蛋白和脂质相的观察。 因此，这是绝对必要的，这项研究。 如果冷冻断裂显微照片是有意义的， 解释，这是最好的样品是uncryoprotected，但 冻结得如此之快，以至于在形成之前发生玻璃化 破坏超微结构的大冰晶。 高 压力冷冻机，它简单地施加压力和冷冻剂 共同降低了避免结冰所需的结冰速度 晶体形成，是唯一的工具，将允许良好的 冷冻发生在未冷冻保护的 sample. 因此，它大大减少了伪影，增强了 有意义的解释副本，并增加了 冷冻断裂实验的效率。

This is a proposal for funds to purchase a freeze fracture instrument and a high pressure freezer. The freeze fracture machine is required for determination of: (1) typed and distribution of proteins in membranes; (2) patterns of vesicle fusion; (3) relationships between extracellular matrix components; and (4) the role of non-bilayer lipids in membrane function. The research proposed includes analysis of natural and artificial membranes, comparison of control and experimental samples, and investigation of plants and protists. (Minor users will also examine fungi, animals, and DNA molecules.) Freeze fracture, which involves cleaving and replicating cells and membranes, is the only technique that allows structural observation of intramembrane proteins and lipid phases. Therefore, it is absolutely required for this research. If freeze fracture micrographs are to be meaningfully interpreted, it is best for the sample to be uncryoprotected, but frozen so fast that vitrification occurs before the formation of large ice crystals that destroy ultrastructure. The high pressure freezer, which briefly applies pressure and cryogen together to lower the rate of freezing required to avoid ice crystal formation, is the only instrument that will allow good freezing to occur below the surface layer of an uncryoprotected sample. Therefore, it greatly reduces artifacts, enhances meaningful interpretation of replicas, and increases the efficiency of freeze fracture experimentation.