Life in 3D, expanding the structural length scales via Serial Block Face Scanning Electron Microscopy (SBF-SEM)

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Bioimaging is one of the key technologies for life science research but the way life functions has however often been studied by reducing this to 2-dimensional projections for instance by light and / or electron microscopy. Although this approach has led and will also in the future lead to breakthrough discoveries there is a realisation that if possible "life" should be studied as much as possible in a 3D environment, ideally in the context of a tissue or organism. Some of the newer microscopy techniques therefore are focussing on this aspect and allow for imaging large volumes in all 3 dimensions, e.g. by using light sheet light microscopy. Electron Microscopy has generally focussed on higher resolution structural information but alongside the "resolution revolution" in structural cryo Electron Microscopy (EM) which is revealing ever greater molecular details, there has been a quieter but probably equally important revolution in volume EM. By sequentially scanning the block face of an embedded sample and removing the scanned top layer, a large high-resolution 3-dimensional volume can be acquired. Although at somewhat lower resolution this volume is much larger than possible by standard (serial thick section) Electron Tomography. This allows for the structural visualisation of complete cells and the interactions of these cells with other cells or the extracellular matrix inside a tissue. As all life is built in 3 Dimensions, Serial Block Face Scanning Electron Microscopy (SBF-SEM) has now become the method of choice for a growing number of life science studies.We would like to apply for a Scanning Electron Microscope (SEM) with an integrated diamond knife based microtome. We will integrate the technology into the Wolfson Bioimaging Facility, the centralised microscopy facility of the Faculty of Biomedical Sciences at the University of Bristol which houses both light and electron microscopy and is well-known for its Correlative Microscopy technologies.We will apply the tool to study a variety of research questions that can only be studied in 3D. These include but are certainly not limited to the formation of synapses in the brain, the infiltration of macrophages into tissue, and the formation and release of platelets in the blood stream. Importantly this tool will allow us to also study the interaction of the extracellular matrix with cells, an area where 3D imaging is critical.Last but not least we will use the microscope to advance our understanding in the Synthetic Biology field, one of the highlight areas of the BBSRC. Some of the Synthetic Biology research in Bristol focusses on the development of artificial extracellular supports with the idea to grow cells on, and ultimately to use these for transplantation. In order to study the interaction of seeded cells with such matrix supports the SBFSEM will be of critical importance.Training of the next generation of scientists is an important aspect within the Wolfson Bioimaging Facility. As home of an international EMBO course and other national training courses the technology would be of special interest to seamlessly integrate into some of those training courses, providing students with training in the latest state of the art technology.