High pressure freezing to capture secretory pathway dynamics

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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.