High resolution of SEM imaging of meristems parasites bacteria tissues sperm and biomaterials in a multi-user facility

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We need a new high resolution SEM to provide high quality SEM imaging for a wide range of projects across the department. The new SEM will also allow the development of correlative microscopy approaches, broadening further its applications. Wide access to the new SEM for Biology Department researchers will be ensured by locating it in the Imaging and Cytometry Laboratory of the Technology Facility (TF). The TF has established the necessary infrastructure and support mechanisms to ensure easy access, training and maintenance. The TF location will also allow access by external academic and commercial research groups. Examples of projects that require the new SEM are listed below:- 1. Regulation of auxillary meristem activity (Professor Ottoline Leyser) Professor Leyser¿s research centres on the hormonal control of shoot branching in Arabidopsis. Shoot branches arise from axillary meristems (Ams) that are laid down in the axils of the leaves initiated by the primary shoot meristem. Leyser¿s group uses mutants defective in AM regulation, in combination with hormonal treatments to unravel the network of genes and signals that control AM activity. Arabidopsis AMs consist of a dome of cells approximately 50¿m across, and thus SEM provides the ideal way to examine their structure and the early events in their ontogeny. 2. Tissue engineering scaffolds (Professor Jenny Southgate) Professor Southgate will progress her studies of tissue engineered scaffolds developed from natural decellularised and synthetic materials. These will be imaged to analyse surface topography and pore size and to assess the attachment, organisation, morphology and differentiation of cells maintained on scaffolds in three-dimensional cultures. 3. Oviduct-sperm interatctions (Professor Henry Leese) Professor Leese¿s group works on sperm binding to bovine oviduct epithelial cells. The current SEM is used to determine the degree of differentiation of cultured bovine oviduct epithelial cells following on from previous work (3-6) and the numbers of sperm which bind following co-incubation. We will be able to use the more versatile cryostage to capture various time points on an improved dynamic resolution compared to our current methodology and analysis time will be greatly improved with the digital imaging capabilities. 4. Infections parasites (Professor Deborah Smith) Professor Smith¿s group uses molecular genetic and cell biological approaches to study host-parasite interaction in several species of protozoan parasite. High and low level resolution microscopy are essential tools for this research. Access to a SEM facility will allow additional phenotypic characteristics to be accurately assessed, particularly with respect to surface perturbation of infective parasite stages and the dynamics of host cell entry e.g. during phagocytosis of Leishmania species. 5. Development of correlative microscopy Dr O¿Toole is currently seeking better methodologies to enable correlation between fluorescence and electron microscope images. Preliminary studies with Dr Chong on Archaea with the proposed SEM have shown the ability to gain significant advances in the information that can be obtained from coupling these techniques together. We have demonstrated that it is possible to correlate internal fluorescence structure with the detailed extracellular morphology. 6. Jeol UK Collaboration This collaboration aims to develop Quantomix capsule applications to allow surfaces of live cells to be imaged with the SEM. The true capabilities of this are still being elucidated, but we hope to progress these studies significantly through the proposed collaboration. 7. Avacta Ltd The new SEM will also be exploited by a local SME, Avacta, who are providing both a financial contribution to the purchase along with in-kind support for future training initiatives. This will significantly enhance Avacta¿s research effort as well as strengthening our on-going collaborative work with the