Dual Beam Environmental Scanning Electron Microscopy for Novel Applications in the Life Sciences

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We are seeking funds to purchase a Quanta 3D electron microscope. This instrument is a dual beam instrument, combining a focused ion beam (FIB) and an environmental scanning electron microscope. It opens up a wide range of novel applications across the BBSRC remit, but it is a very young technology and as such this project will also address itself to developing optimised protocols for sectioning different types of samples with the ion beam and understanding the impact of gallium ion implantation and consequences for subsequent imaging. Three broad classes of experiments will be performed: (1) we will be developing methods for cutting through samples including hard/soft interfaces (2) we will develop cryoFIB approaches to permit imaging of sectioned samples, such as cells which have been frozen to keep cell contents intact (3) we will perform serial sectioning and imaging to permit 3-dimensional reconstruction of complex samples, at a resolution exceeding that available, for instance, by optical sectioning. A wide range of sample types will be examined, using knowledge based on previous work by the three applicants and external collaborations. In particular we will initially be interested, under project 1, in cell growth at hard interfaces in the context of medical devices and bioreactors and biofilms, and in sectioning whole wheat grains. In project 3, we will be exploring porosity in tissue scaffolds and compacted drug formulations, and the location of labeled organelles and proteins within cells. As we gain understanding of the underlying physics, permitting us to optimise sectioning strategies, we will ensure dissemination to the wider community to facilitate wider use of this instrumentation.