High pressure freezing to capture secretory pathway dynamics

拨款申请分为两部分。首先，该提案是为继续支持植物科学界高压冷冻设施的运营提供资金。这是由BBSRC资助的设备，允许通过在高压下冻结来非常快速地保存用于电子显微镜的生物材料。这意味着避免了化学保存造成的人工制品，也避免了冰晶对组织的破坏。一些科学家目前正在使用该设施，并需要继续这样做，以完成他们的研究方案。第二，申请资金支持电子显微镜，这是支持牛津布鲁克斯大学植物细胞生物学研究碳水化合物生产和蛋白质加工所涉及的亚细胞系统(分泌途径)的功能所必需的。一种名为电子断层扫描的新技术将被用来计算出参与这些过程的细胞隔间的三维组织。同时，通过高压冷冻保存的材料将与抗体结合作为染色，以便在细胞内定位与分泌途径内结构的组织和功能有关的特定蛋白质。

The grant application is two fold. Firstly the proposal is for funding to continue supporting the running of a high-pressure freezing facility for the plant science community. This is BBSRC funded equipment that permits the very rapid preservation of biological material for electron microscopy by freezing under high pressure. This means that artefacts caused by chemical preservation are avoided and also ice crystal damage to tissue is avoided. A number of scientists are currently using the facility and need to continue to do so in order to complete their research programmes. Secondly, funding is requested to support the electron microscopy which is required to support the Oxford Brookes plant cell biology research into the functioning of the sub-cellular system (the secretory pathway) involved in carbohydrate production and protein processing. A new technique, electron tomography will be used to work out the three-dimensional organisation of the cell compartments involved in these processes. At the same time material preserved by high/pressure freezing will be used in combination with antibodies as stains, in order to locate, within the cell specific proteins that are involved in the organisation and functioning of the structures within the secretory pathway.