Liverpool 3View: a national hub for 3D-EM bioscience research

New 3View equipment is required for imaging the fine details of the insides of cells and tissues in 3D. In all of our projects we are using multidisciplinary approaches to address our questions however they are all united by the need for imaging at high resolution. The challenge with this is that the closer you look the harder it is to see the surrounding context. The solution is to take images of many serial sections through the sample and then digitally reconstruct them into a 3D volume that can be viewed from any angle. This helps with understanding the spatial relationships between cells within tissues, between subcellular compartments and for observing rare events. The 3View system makes this type of analysis routine, allowing rapid processing of samples.
Our projects span a broad range of subjects from basic cell biology and physiology such how cells organise their communication systems when they are moving, how the recycling machinery of the cell works, changes to cells and tissues during ageing and the biogenesis and structural organisation of healthy tissues. Our work also extend through to translational projects, for example interactions between engineered stem cells and host tissue, development of novel cell scaffolds for treating age-related eye disorders, infection routes taken by parasites or bacteria in the gut and the development of novel nanoparticle tools for delivering cargoes to cells.
The equipment will be housed in a leading UK electron microscopy unit and available to external scientists from the UK and Europe. 3View is a new technology that is only beginning to become established within the UK. As part of making Liverpool a national hub for this technology and to help drive innovative applications, we will set up a workshop programme that will act as a forum for experienced users and will enable new users to learn about sample processing, imaging and analysis.

We would like to purchase a Gatan 3View2XP system to enable high-resolution 3D-electron microscopy (EM) of cell and tissue volumes to be routinely performed. Our proposals will improve understanding of: i) cell signalling integration and compartmentalisation; ii) organelle biogenesis, structure and interactions; iii) aging related changes in model organisms and human tissue; iv) stem cell engineering for translational applications; v) host-parasite and gut-bacteria interactions required for infection; vi) cell-matrix and musculoskeletal biology in humans and animals; vii) cellular interactions of novel biofunctionalised nanoparticle reagents.
The projects described represent multidisciplinary approaches to address the research questions. The common theme within all of the projects is the need for 3-D EM imaging of cell and tissue volumes to allow visualisation of subcellular compartments, cell-cell and cell-substrate interactions, tissue morphology and organism anatomy. The proposal incorporates a consortium of local and external academics from leading UK research institutes with immediate requirements for access to this new technology. Since we are a leading bio-EM unit within the UK with a critical mass of high quality projects, we are well positioned to extend our capabilities to become one of the national hubs for this technology with reserved capacity for external groups.

The proposed projects will impact the following groups over the indicated timescales
- Local and external researchers that will access the equipment (immediate impact)
- Scientists within & outside research field of the PIs (months, years: Detailed in Academic Beneficiaries)
- Medical and veterinary clinicians (months, years)
- Biotechnology companies (years)
- General public (months, years)

3View is a new technology that was initially developed and established in Germany followed by a rapid uptake of installed capacity in the US and the Far East. The UK has an international reputation in advanced imaging capabilities and research and 3View is highly complementary to many of these approaches. However, we currently lack national 3View capacity to serve existing demand with consequent risks to scientific competitiveness. A strategic investment in this technology to improve UK capacity has the potential to benefit any groups that use imaging in their research. There will be a specific benefit to BBSRC-funded researchers. All current 3View systems have been funded by the MRC, the Wellcome Trust or CRUK. We propose to reserve a minimum of 20% of our capacity for external users with priority given to those with BBSRC funded projects.
All users of the equipment will be offered full training in advanced skills of sample preparation, imaging and image processing. We will also establish a workshop that will provide a national focal point for new and experienced users. The workshop will involve leaders in 3View imaging from the UK and Europe and facilitate training, discussion of best practice, troubleshooting and provide networking opportunities. Together, these initiatives will contribute to the professional development of all users of our facility and help to promote co-operation amongst the network of 3View facilities within the UK.
Scientifically, our application is focussed on our strengths in cell biology, physiology, biomedical and veterinary research. The work that will use the 3View includes highly translational studies, for example, the investigations into bacterial-gut interactions and the role of dietary factors at modulating this (Campbell). Advances in this area have potential human quality of life and health benefits and may inform government policy and industry protocols for example in the types of additives used in food. Similarly, the work that will be done to understand the mechanisms of invasion of Trypanosomes across the Tsetse fly gut wall (Acosta-Serrano) has the potential to reveal new therapeutic avenues over the next decade for treating this debilitating disease. Research into age-related deterioration of tendon mechanical function (Laird & Clegg) may provide insights into methods of limiting injury risk that will inform treatment practices and drug development. Other examples with potential to have medical benefits and biotechnology company involvement include: novel nanotechnologies for drug delivery (Brust and Jones), developing new stem cell tools for treating kidney disease (Wilm & Murray) and transplants for treating age-related eye disorders (Williams).
The Biomedical EM unit and all participating co-applicants are committed to engaging with the public about their research. These include visits to the facility and labs by schools, fundraisers and patients; producing websites written in a lay style, publicising the results of our research via local and national press with support from funding agency/University press offices and participating in educational forums.