Development of light and electron microscopy facilities at University of Sheffield. (360G-Wellcome-093134_Z_10_Z)

In order to understand biological processes, it is necessary to be able to visualize individual components in their natural cellular, tissue or organismal environment. The aim of this proposal is to facilitate such visualization by developing new electron microscopy approaches within the Electron Microscopy (EM) Facility and maintaining and further developing the Wellcome-funded Light Microscopy Facility (LMF) at the University of Sheffield. The LMF is a multiuser facility that provides access to confocal microscopy which is widely used to study biological processes in living and fixed tissues. Our existing confocals are heavily used and usage is predicted to increase in the next year. One instrument is greater than eleven years old and one aim of this application is to replace it so that our users' requirements may be accommodated. Our current EM facility allows the sectioning of tissues and cells embedded in resin and sectioned at room temperature. While this provides us with usefu l ultrastructural details of cells, it is not usually compatible with antibody-labelling of molecular components. We are therefore requesting funds for freeze substitution equipment that will enable us to perform immunogold labelling of specific proteins in samples with well-preserved morphology.